Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Data Requirement: : EPA DP Barcode D288775 EPA MRID 458677 10 EPA Guideline 70 1( Special Study)

Test material: Purity: not reported Common name: Atrazine Chemical name: IUPAC CAS name 6 chloro N ethyl N'( 1 methylethyl) 1,3,5 triazine 2,4 diamine CAS No. 1912 24 9 synonyms EPA PC Code: 80803 Primary Reviewer: Thomas M. Steeger, Ph. D., Senior Biologist Date: April 12, 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 Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 2 of 31 CITATION: Smith, E. E., L. DuPreez, and K. Solomon. 2003. Gonadal and laryngeal responses to field exposure of Xenopus laevis to atrazine in areas of corn production in South Africa. The Institute of Environmental and Human Health, Texas Tech University, Lubbock, TX and School of Environmental Sciences and Development, Potchefstroom University for CHE, Potchefstroom, South Africa. Sponsor: Syngenta Crop Protection, Inc., Laboratory Study ID: ECORISK Number SA 01C.

EXECUTIVE SUMMARY:

This report is part of a multiphase study examining the effects of atrazine on African clawed frogs ( Xenopus laevis) in their native habitat ( South Africa). This phase of the study summarizes the body weights, lengths and condition index ( weight/ length) of both male and female collected from reference site ( no corn grown and no atrazine/ triazine use) and experimental site ( corn grown and atrazine/ triazines used) ponds. The study also examined several aspects of gonadal growth ( testis weight, gonadosomatic index) and development ( incidence of deformities, cellular morphology) in frog testes collected from the two sites to determine whether atrazine exposure had affected these measurement endpoints.

While males were smaller than females at both study sites, there was no difference in weight or length of males or females collected at reference or experimental sites. Condition indices for males and females did not differ between reference and experimental collection sites. However, the mean testes weight from reference ponds ( 0.037 ± 0.021 mg) was statistically different from mean weight of testes ( 0.046 ± 0.018 mg) from frogs collected at experimental sites. GSI was significantly different for males collected from reference ( 0.13 ± 0.04) compared to males collected from experimental sites ( 0.17 ± 0.04).

There were no statistical differences in the cellular morphology of testes collected at reference sites from those collected at experimental site. The incidence of external abnormalities ( missing limbs) was 1% across all animals collected ( reference and experimental) and was attributed to the predatory effects of turtles. The incidence of testicular oocytes was 3% in males collected from reference ponds and 2% in experimental ponds. The study claims that laryngeal somatic index for males and females from reference and experimental sites were not significantly different ( p > 0.05); however, no raw data were provided to verify these observations.

Although not discussed at length in this phase of the study, the interpretation of these data is complicated by the fact that atrazine, its degradates and other triazines were detected at significant levels in reference ponds. Additionally, at some sites, frogs were collected over a protracted period of time ( 6 months), rendering GSI a questionable metric. This study provides evidence that under the conditions studied, the incidence of testicular oocytes in Xenopus laevis is roughly 2 to 3%. Because of the presence of atrazine in both sampling areas, a regression based analysis would be more appropriate than hypothesis testing based analysis. Unfortunately, though, the study is not designed for regression analysis. Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 3 of 31 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. ° Test material identity, strength, purity and composition were characterized ° Independent QA inspections of raw data and final report were conducted. ° Interim Report was written ° Raw data, documentation, records, protocols, and final phase report will be archived.

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 were field collected in two areas ( non corn growing areas and corn growing areas) in the vicinity of Potchefstrooom, South Africa, using traps baited with liver and meat scraps. Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 4 of 31 B. STUDY DESIGN:

Objective: 1. To determine if exposure to triazines used in corn production affects the incidence or extent of morphological changes in the gonads or intersex gonads in the male and female metamorphs and adults of X. laevis. 2. To determine whether there are changes in the morphology of the male larynx in metamorphs and adults of X. laevis in response to exposures to triazines used in corn production. 3. To determine if there are changes in plasma steroid concentrations of adult X. laevis in response to exposures to triazines used in corn production. 4. To determine if other frogs present at the sites show any morphological or other changes in the gonads. 5. To assess feasibility of determining if X. laevis in the exposed sites have been subjected to selection pressure.

1. Experimental Conditions Sampling was conducted in April and May approximately 6 months after atrazine was applied ( October / November); water samples to characterize exposure were collected one month prior to frog sampling. Environmental parameters that were measured at each site included habitat description ( vegetative cover and water depth). Water samples were analyzed for atrazine, related triazines, other pesticides and metals.

A total of eight sites were sampled in two adjacent regions ( Viljoenskroon corn growing region = E; non corn growing region = C) in the vicinity of Potchefstroom, South Africa.

Compounds of interest included atrazine, its metabolites desethylated atrazine ( DEA), desisopropyl atrazine ( DIA), diaminochlorotriazine ( DACT), plus other herbicides ( terbutylazine, simazine and acetochlor).

Frogs were sampled after the rainy season in April and May; however, at atrazine sites E1 and E8, frogs were collected from April to June and from April to September, respectively, due to low sample sizes. The low sample sizes at sites E1and E8 were attributed to the introduction of sharptooth catfish ( Clarius gariepinus) into the ponds due to high runoff. Sampling at all other sites occurred once or twice within a " few days." Frogs were collected in 10 baited traps and harvested after two days in " clean water". Afterwards, animals were housed individually in 2 L plastic containers for 48 hours to recover from " capture stress".

Total larynx weight was used to determine the response of the larynx in animals from experimental and control study sites. Adult males from each site were selected randomly for a total of 170 animals.

Testes were processed for routine paraffin embedding, serially sectioned at 5

m, and stained with hematoxylin and eosin. Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 5 of 31 Three slides were selected: slide A was selected from the rostral terminus of the testes on the basis of the first observation of a seminiferous tubule; slide B was selected from the middle; and slide C was selected from a similar arbitrarily assigned caudal region of the testis with a complete seminiferous tubule. Three fields were photographed from each of the three slides yielding nine photomicrographs of each testis. The photomicrographs were overlain with a grid and the cell/ tissue type under each crossbar was identified and counted by three different readers to determine the representative fractional volume of spermatogonia, spermatocyte, sperm, connective tissue, blood vessel " other" morphological features.

For oocytes, every 200th micron section was evaluated.

II. RESULTS and DISCUSSION: [ All results discussed in this section and the next are those reported by the study authors. Although supplemental data are typically used in a qualitative manner only, EFED verified spreadsheet data and ran basic statistical analyses on the major study parameters. See attached appendix. If results differed in any substantive way, the difference was reported in the text below.]

The researchers were unable to collect Stage 66 metamorphs except in reference site C3; therefore, the study focused on adults. According to the report, the targeted number of fish, i. e., 20 males and 20 females per site, could not be achieved ( Table 1) at sites E1 and E8 due to the introduction of sharptooth catfish; however, no explanation is provided as to why the targeted number could not be obtained from reference site C3.

Table 2 presents the mean body weight and snout vent lengths ( ± standard deviation) of frogs collected in reference and experimental sites. There was no statistical difference in the average SVL of females collected from reference sites ( 71.8 ± 4.45 mm) compared with females collected from experimental sites ( 73.9 ± 10.1 mm). Additionally, there was no difference in mean weight of females from reference sites ( 36.5 ± 4.32 g) and experimental sites ( 42.8 ± 15.7 g).

There was no statistical difference in the average SVL of males collected from reference sites ( 62.4 ± 8.45 mm) compared with males collected from experimental sites ( 62.6 ± 2.87 mm). There was also no statistical difference in mean weight of males collected at reference ( 25.8 ± 9.93 g) and experimental sites ( 27.0 ± 4.14 g).

While the report states that there was no difference in weight of the left testis from reference ( 18.6 mg) and experimental sites ( 23.3 mg) nor between the weight of the right testis from reference ( 15.8 mg) and experimental ( 22.1 mg) site collected animals, there were differences in the total testicular weight ( combined right and left testis) between reference and experimental collection sites. The mean testes weight from control ( 0.037 ± 0.021 mg) was statistically different ( ANOVA, p < 0.0449) than mean weight of testes ( 0.046 ± 0.018 mg) from frogs collected at experimental sites.

Table 2. Mean snout vent length and body weight of male and female adult frogs collected at experimental ( E1  E8) and reference ( R1  R3) sites.

Pond Snout Vent Length ( mm) Body Weight ( g)

Males Females Males Females E1 59.20 ± 9.49 64.48 ± 14.3 23.39 ± 14.4 29.15 ± 19.2 Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 6 of 31 E3 66.63 ± 6.99 74.39 ± 6.94 32.57 ± 8.99 39.38 ± 8.47 E4 63.83 ± 3.35 77.54 ± 10.9 29.90 ± 4.17 48.82 ± 18.5 E6 62.66 ± 9.58 88.50 ± 9.53 26.11 ± 12.4 66.96 ± 22.7 E8 60.72 ± 7.28 64.47 ± 13.8 23.09 ± 8.79 29.80 ± 18.9 R1 58.00 ± 6.64 69.55 ± 8.02 19.44 ± 6.10 32.83 ± 11.1 R3 56.99 ± 10.9 68.91 ± 14.5 20.75 ± 13.3 35.35 ± 20.2 R8 72.11 ± 7.19 76.95 ± 7.04 37.26 ± 12.0 41.25 ± 10.4 Average condition indices ( body weight ÷ snout vent length) for females collected from reference sites ( 0.48 ± 0.15 g/ mm) were not significantly different ( p> 0.0562) from females collected at experimental sites ( 0.54 ± 0.20 g/ mm). Similarly, male CI between reference ( 0.39 ± 0.14 g/ mm) and experimental sites ( 0.42 ± 0.12 g/ mm) were not statistically different ( p> 0.237).

The report states that mean gonadalsomatic indices ( GSI = total weight of gonad ÷ body weight) were lower for males collected in reference sites than in experimental sites but were not statistically different. EFED's analysis ( see attached SAS output), though, indicates that GSI were significantly different ( ANOVA, p < 0.0001) for males collected from reference ( 0.13 ± 0.04) sites compared to males collected from experimental sites ( 0.17 ± 0.04).

Morphological abnormalities ( deformed limbs) were observed in 3 out of 212 ( 0.01%) of the animals and were attributed to attacks by the Cape terrapin ( Pelomedusa subrufa).

Based on gross morphology, only 2 out of 101 males examined had discontinuous gonads, and 4 had testes of unequal size.

The extent of morphological changes in adult frog testes was determined by evaluating 56 testes from reference sites and 43 from experimental sites ( Table 3). There were no statistical differences in the cell types comprising testes collected from reference and experimental sites.

Testicular oocytes were observed in 3% of the animals collected from reference sites and 2% of the frogs collected from experimental sites.

The study claims that laryngeal somatic index for males and females from reference and experimental sites were not significantly different ( p > 0.05); however, no raw data were provided to verify these observations. Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 7 of 31 Table 3. Mean cell type identified in testes of male frogs collected from reference ( C1, C3 and C6) and experimental ( E1, E3, E4, E6 and E8) sites along with associated t test p values.

Site Spermatogoni a Spermatocyte s Sperm Connective Tissue and Other Cells Blood Vessels C1 3.0 25.6 55.9 14.9 0.5 C3 4.5 34.3 46.3 14.6 0.3 C6 2.8 22.6 60.3 13.9 0.3 E1 3.4 28.8 53.4 13.7 0.8 E3 3.6 24.9 58.1 13.2 0.2 E4 3.0 24.5 59.8 12.6 0.2 E6 3.2 20.8 62.4 13.2 0.3 E8 2.9 30.1 52.2 14.5 0.2 Mean of C1  C6 3.5 27.5 54.2 14.5 0.4 Mean of E1  E8 3.2 25.8 57.2 13.5 0.3 t test p values 0.6214 0.6382 0.4782 0.0754 0.7305 C. REPORTED STATISTICS: Statistical analyses of frog numbers at each site were conducted using standard methods such as Chi square, Kruskal Wallis and Student's t test. The initial analyses of histopathology were carried out using standard ANOVA procedures followed by Tukey's multi comparison test using Minitab ® statistical software.

D. VERIFICATION OF STATISTICAL RESULTS: Statistical analyses conducted using SAS ( Statistical Analysis System, Release 8.01, Cary, North Carolina). See attached printout.

E. STUDY DEFICIENCIES:

Atrazine and/ or triazine degradates were present in control sites. Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 8 of 31 While the low sample size at sites E1 and E8 may have been explained by the sudden introduction of predatory catfish, it doesn't explain why the remaining corn growing sites ( E3, E4 and E6) also yielded fewer frogs than non corn growing sites.

Because frogs were sampled over disproportionate period of time, e. g., 6 months for site E8, animals were at different stages of development.

Frog traps were baited with liver/ meat scraps that may have contained hormones.

Following capture, frogs were housed individually in " clean water" for 48 hours. The volume and characteristics of this water were not described in the study.

F. REVIEWER'S COMMENTS:

Atrazine is typically applied in October / November when corn is planted in South Africa. Frog sampling was conducted after the rainy season in April to May, and water samples to characterize exposure were collected within a time span of four weeks before frogs were sampled. At exposure site E8, frogs were sampled over a six month time frame; the authors attributed the fewer number of frogs to a high runoff event that washed catfish into the pond and the catfish preyed on the frogs. Given the aerial photos and the fact that the majority of the ponds relied on rain ( runoff) and were not dependent on a stream, it is unclear where the catfish came from. Because Clarius is an aquaculture species, it is possible that aquaculture ponds were in the vicinity of E1 and E8.

Frogs were attracted into baited traps containing liver and/ or scrap meat that may have contained hormones. Apparently trapped animals were held in cages for two days which would likely have provided sufficient time to consume all of the bait. Also, depending on catch rates, the loading within the cages could have stressed the frogs. Although it doesn't state the desired sample size in the report's methodology, apparently ( based on the raw data), 20 male and 20 female frogs were to be sampled at each site. Table 1 shows that while the desired sample size was generally available for frogs collected in non corn growing areas, sample sizes were minimally 50% lower in corn growing areas. In site E1, sampling for female frogs extended into late June and at site E8 sampling extended into mid August for females and into mid September for males. Extended sampling periods at these two sites may have resulted in collecting animals that were at considerably different states of development. The gonadosomatic index and ovarian aromatase activity in females collected from sites E1 and E8 expressed the highest variability relative to any of the other treatment sites.

Table 1. Summary of total number of Xenopus laevis collected from April through May 2002 at non corn growing ( C) and corn growing ( E) sites in South Africa.

Site Males Females C1 20 ( Apr 21  Apr 23) 20 ( Apr 21  May 1)

C3 17 ( Apr 4  Apr 11) 20 ( Apr 4  Apr 11) Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 9 of 31 C6 20 ( May 13  May 15) 20 ( May 13  May 14)

E1 8 ( April 29  May 7) 8 ( April 29  June 29)

E3 10 ( Apr 9  Apr 17) 10 ( Apr 9  Apr 18)

E4 10 ( Apr 13  Apr 15) 10 ( Apr 14  Apr 16)

E6 10 ( May 5  May 9) 10 ( May 5  May 6)

E8 6 ( Apr 29  Sep 17) 15 ( Apr 29  Aug 17)

G. CONCLUSIONS:

This report is part of a multiphase study examining the effects of atrazine on African clawed frogs ( Xenopus laevis) in their native habitat ( South Africa). This phase of the study summarizes the body weights, lengths and condition index ( weight/ length) of both male and female collected from reference site ( no corn grown and no atrazine/ triazine use) and from experimental site ( corn grown and atrazine/ triazines used) ponds. The study also examined several aspects of gonadal growth ( testis weight, gonadosomatic index) and development ( incidence of deformities, cellular morphology) in frog testes collected from the two sites to determine whether atrazine exposure had affected these measurement endpoints.

While males were smaller than females at both study sites, there was no difference in weight or length of males or females collected at reference or experimental sites. Condition indices for males and females did not differ between reference and experimental collection sites. However, the mean testes weight from reference ponds ( 0.037 ± 0.021 mg) were statistically different than mean weight of testes ( 0.046 ± 0.018 mg) from frogs collected at experimental sites. GSI was significantly different for males collected from reference ( 0.13 ± 0.04) compared to males collected from experimental sites ( 0.17 ± 0.04).

There were no statistical differences in the cellular morphology of testes collected at reference sites and those collected at experimental site. The incidence of external abnormalities ( missing limbs) was 1% across all animals collected ( reference and experimental) and was attributed to the predatory effects of turtles. The incidence of testicular oocytes was 3% in males collected from reference ponds and 2% in experimental ponds. The study claims that laryngeal somatic index for males and females from reference and experimental sites were not significantly different ( p > 0.05); however, no raw data were provided with which to verify these observations.

Although not discussed at length in this phase of the study, the interpretation of these data is complicated by the fact that atrazine, its degradates and other triazines were detected at significant levels in reference ponds. Additionally, at some sites, frogs were collected over a protracted period of time ( 6 months) rendering GSI a questionable metric. This study provides evidence that under the conditions studied, the incidence of testicular oocytes in Xenopus laevis in South Africa is roughly 2 to 3%. Because of the presence of atrazine in both sampling areas, a regression based analysis would be more appropriate than hypothesis testing based analysis. This study, though, was not designed for regression analysis. Can you include a summary conclusion Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 10 of 31 statement here? Anything useful in this study? After the study is completed, a more definitive conclusion will be reached.

MEAN MALE SVL FOR REFERENCE ( R) AND EXPERIMENTAL ( E) COLLECTION SITES 109 Obs POND _ TYPE_ _ FREQ_ MEAN STD 1 E1 0 8 59.2000 9.4870 2 E3 0 10 66.6300 6.9863 3 E4 0 10 63.8300 3.3549 4 E6 0 10 62.6600 9.5755 5 E8 0 6 60.7167 7.2841 6 R1 0 20 58.0000 6.6373 7 R3 0 17 56.9882 10.9170 8 R6 0 20 72.1100 7.1908 Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 11 of 31 ANOVA FOR MALE SLV BETWEEN SITES 110 The ANOVA Procedure Class Level Information Class Levels Values SITE 2 C E Number of observations 101 Dependent Variable: LENGTH Sum of Source DF Squares Mean Square F Value Pr > F Model 1 2.016502 2.016502 0.02 0.8822 Error 99 9041.101320 91.324256 Corrected Total 100 9043.117822 R Square Coeff Var Root MSE LENGTH Mean 0.000223 15.22363 9.556373 62.77327 Source DF Anova SS Mean Square F Value Pr > F SITE 1 2.01650201 2.01650201 0.02 0.8822 Levene's Test for Homogeneity of LENGTH Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F SITE 1 77939.9 77939.9 6.28 0.0139 Error 99 1229054 12414.7 Bartlett's Test for Homogeneity of LENGTH Variance Source DF Chi Square Pr > ChiSq SITE 1 5.1814 0.0228 Dunnett's t Tests for LENGTH NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 99 Error Mean Square 91.32426 Critical Value of Dunnett's t 1.98422 Comparisons significant at the 0.05 level are indicated by ***.

Difference SITE Between Simultaneous 95% Comparison Means Confidence Limits E  C 0.2850  3.5203 4.0902 Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 12 of 31 NONPARAMETRIC COMPARISON OF MALE LENGTH BETWEEN SITES 114 The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable LENGTH Classified by Variable SITE Sum of Expected Std Dev Mean SITE N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ C 57 2872.50 2907.0 145.999621 50.394737 E 44 2278.50 2244.0 145.999621 51.784091 Average scores were used for ties.

Wilcoxon Two Sample Test Statistic 2278.5000 Normal Approximation Z 0.2329 One Sided Pr > Z 0.4079 Two Sided Pr > | Z| 0.8159 t Approximation One Sided Pr > Z 0.4082 Two Sided Pr > | Z| 0.8163 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 0.0558 DF 1 Pr > Chi Square 0.8132 Median Scores ( Number of Points Above Median) for Variable LENGTH Classified by Variable SITE Sum of Expected Std Dev Mean SITE N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ C 57 29.0 28.217822 2.503874 0.508772 E 44 21.0 21.782178 2.503874 0.477273 Average scores were used for ties.

Median Two Sample Test Statistic 21.0000 Z  0.3124 One Sided Pr < Z 0.3774 Two Sided Pr > | Z| 0.7547 Median One Way Analysis Chi Square 0.0976 DF 1 Pr > Chi Square 0.7547 Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 13 of 31 MEAN FEMALE SVL FOR REFERENCE ( R) AND EXPERIMENTAL ( E) COLLECTION SITES 116 Obs POND _ TYPE_ _ FREQ_ MEAN STD 1 E1 0 6 64.4833 14.3272 2 E3 0 10 74.3900 6.9437 3 E4 0 10 77.5400 10.9218 4 E6 0 10 88.5000 9.5274 5 E8 0 15 64.4733 13.8195 6 R1 0 20 89.5450 91.3941 7 R3 0 20 68.9100 14.4936 8 R6 0 20 76.9500 7.0433 Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 14 of 31 ANOVA FOR FEMALE SLV BETWEEN SITES 117 The ANOVA Procedure Class Level Information Class Levels Values SITE 2 C E Number of observations 111 Dependent Variable: LENGTH Sum of Source DF Squares Mean Square F Value Pr > F Model 1 628.8675 628.8675 0.38 0.5363 Error 109 178119.4467 1634.1234 Corrected Total 110 178748.3142 R Square Coeff Var Root MSE LENGTH Mean 0.003518 52.99887 40.42429 76.27387 Source DF Anova SS Mean Square F Value Pr > F SITE 1 628.8675382 628.8675382 0.38 0.5363 Levene's Test for Homogeneity of LENGTH Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F SITE 1 1.864E8 1.864E8 0.83 0.3656 Error 109 2.461E10 2.2581E8 Bartlett's Test for Homogeneity of LENGTH Variance Source DF Chi Square Pr > ChiSq SITE 1 70.8565 <. 0001 Dunnett's t Tests for LENGTH NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 109 Error Mean Square 1634.123 Critical Value of Dunnett's t 1.98197 Comparisons significant at the 0.05 level are indicated by ***.

Difference Simultaneous SITE Between 95% Confidence Comparison Means Limits E  C  4.776  20.036 10.483 Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 15 of 31 NONPARAMETRIC COMPARISON OF FEMALE LENGTH ACROSS SITES 121 The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable LENGTH Classified by Variable SITE Sum of Expected Std Dev Mean SITE N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ C 60 3256.0 3360.0 168.985548 54.266667 E 51 2960.0 2856.0 168.985548 58.039216 Average scores were used for ties.

Wilcoxon Two Sample Test Statistic 2960.0000 Normal Approximation Z 0.6125 One Sided Pr > Z 0.2701 Two Sided Pr > | Z| 0.5402 t Approximation One Sided Pr > Z 0.2707 Two Sided Pr > | Z| 0.5415 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 0.3788 DF 1 Pr > Chi Square 0.5383 Median Scores ( Number of Points Above Median) for Variable LENGTH Classified by Variable SITE Sum of Expected Std Dev Mean SITE N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ C 60 27.0 29.729730 2.637040 0.450000 E 51 28.0 25.270270 2.637040 0.549020 Average scores were used for ties.

Median Two Sample Test Statistic 28.0000 Z 1.0351 One Sided Pr > Z 0.1503 Two Sided Pr > | Z| 0.3006 Median One Way Analysis Chi Square 1.0715 DF 1 Pr > Chi Square 0.3006 Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 16 of 31 AVERAGE LENGTH OF MALE AND FEMALE FROGS AT REFERENCE AND EXPERIMENTAL SITES 123 Obs SITE SEX _ TYPE_ _ FREQ_ LENGTH SD_ L 1 C F 0 3 78.4683 10.4010 2 C M 0 3 62.3661 8.4536 3 E F 0 5 73.8773 10.0521 4 E M 0 5 62.6073 2.8664 MEAN MALE WEIGHT FOR REFERENCE ( R) AND EXPERIMENTAL ( E) COLLECTION SITES 124 Obs POND _ TYPE_ _ FREQ_ MEAN STD 1 E1 0 8 23.3888 14.4006 2 E3 0 10 32.5740 8.9938 3 E4 0 10 29.9000 4.1673 4 E6 0 10 26.1050 12.3935 5 E8 0 6 23.0917 8.7925 6 R1 0 20 19.4445 6.1022 7 R3 0 17 20.7541 13.2996 8 R6 0 20 37.2590 12.0340 Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 17 of 31 ANOVA FOR MALE WEIGHT BETWEEN SITES 125 The ANOVA Procedure Class Level Information Class Levels Values SITE 2 C E Number of observations 101 Dependent Variable: WEIGHT Sum of Source DF Squares Mean Square F Value Pr > F Model 1 52.00466 52.00466 0.35 0.5572 Error 99 14842.45191 149.92376 Corrected Total 100 14894.45657 R Square Coeff Var Root MSE WEIGHT Mean 0.003492 45.83106 12.24434 26.71624 Source DF Anova SS Mean Square F Value Pr > F SITE 1 52.00466491 52.00466491 0.35 0.5572 Levene's Test for Homogeneity of WEIGHT Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F SITE 1 126859 126859 2.23 0.1387 Error 99 5637162 56941.0 Bartlett's Test for Homogeneity of WEIGHT Variance Source DF Chi Square Pr > ChiSq SITE 1 3.0051 0.0830 Dunnett's t Tests for WEIGHT NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 99 Error Mean Square 149.9238 Critical Value of Dunnett's t 1.98422 Comparisons significant at the 0.05 level are indicated by ***.

Difference Simultaneous SITE Between 95% Confidence Comparison Means Limits E  C 1.447  3.428 6.323 Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 18 of 31 NONPARAMETRIC COMPARISON OF MALE WEIGHT BETWEEN SITES 129 The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable WEIGHT Classified by Variable SITE Sum of Expected Std Dev Mean SITE N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ C 57 2742.0 2907.0 146.005999 48.105263 E 44 2409.0 2244.0 146.005999 54.750000 Average scores were used for ties.

Wilcoxon Two Sample Test Statistic 2409.0000 Normal Approximation Z 1.1267 One Sided Pr > Z 0.1299 Two Sided Pr > | Z| 0.2599 t Approximation One Sided Pr > Z 0.1313 Two Sided Pr > | Z| 0.2626 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 1.2771 DF 1 Pr > Chi Square 0.2584 Median Scores ( Number of Points Above Median) for Variable WEIGHT Classified by Variable SITE Sum of Expected Std Dev Mean SITE N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ C 57 26.0 28.217822 2.503874 0.456140 E 44 24.0 21.782178 2.503874 0.545455 Average scores were used for ties.

Median Two Sample Test Statistic 24.0000 Z 0.8858 One Sided Pr > Z 0.1879 Two Sided Pr > | Z| 0.3757 Median One Way Analysis Chi Square 0.7846 DF 1 Pr > Chi Square 0.3757 Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 19 of 31 MEAN FEMALE WEIGHT FOR REFERENCE ( R) AND EXPERIMENTAL ( E) COLLECTION SITES 131 Obs POND _ TYPE_ _ FREQ_ MEAN STD 1 E1 0 6 29.1450 19.2315 2 E3 0 10 39.3810 8.4697 3 E4 0 10 48.8210 18.5235 4 E6 0 10 66.9560 22.6873 5 E8 0 15 29.8047 18.8819 6 R1 0 20 32.8315 11.1449 7 R3 0 20 35.3455 20.2334 8 R6 0 20 41.2495 10.4261 Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 20 of 31 ANOVA FOR FEMALE WEIGHT BETWEEN SITES 132 The ANOVA Procedure Class Level Information Class Levels Values SITE 2 C E Number of observations 111 Dependent Variable: WEIGHT Sum of Source DF Squares Mean Square F Value Pr > F Model 1 1040.14605 1040.14605 2.97 0.0874 Error 109 38122.22029 349.74514 Corrected Total 110 39162.36634 R Square Coeff Var Root MSE WEIGHT Mean 0.026560 47.58918 18.70147 39.29775 Source DF Anova SS Mean Square F Value Pr > F SITE 1 1040.146048 1040.146048 2.97 0.0874 Levene's Test for Homogeneity of WEIGHT Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F SITE 1 2126150 2126150 5.83 0.0174 Error 109 39770440 364866 Bartlett's Test for Homogeneity of WEIGHT Variance Source DF Chi Square Pr > ChiSq SITE 1 9.1873 0.0024 Dunnett's t Tests for WEIGHT NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 109 Error Mean Square 349.7451 Critical Value of Dunnett's t 1.98197 Comparisons significant at the 0.05 level are indicated by ***.

Difference Simultaneous SITE Between 95% Confidence Comparison Means Limits E  C 6.143  0.917 13.202 Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 21 of 31 NONPARAMETRIC COMPARISON OF FEMALE WEIGHT ACROSS SITES 136 The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable WEIGHT Classified by Variable SITE Sum of Expected Std Dev Mean SITE N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ C 60 3132.50 3360.0 168.996671 52.208333 E 51 3083.50 2856.0 168.996671 60.460784 Average scores were used for ties.

Wilcoxon Two Sample Test Statistic 3083.5000 Normal Approximation Z 1.3432 One Sided Pr > Z 0.0896 Two Sided Pr > | Z| 0.1792 t Approximation One Sided Pr > Z 0.0910 Two Sided Pr > | Z| 0.1820 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 1.8122 DF 1 Pr > Chi Square 0.1782 Median Scores ( Number of Points Above Median) for Variable WEIGHT Classified by Variable SITE Sum of Expected Std Dev Mean SITE N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ C 60 27.0 29.729730 2.637040 0.450000 E 51 28.0 25.270270 2.637040 0.549020 Average scores were used for ties.

Median Two Sample Test Statistic 28.0000 Z 1.0351 One Sided Pr > Z 0.1503 Two Sided Pr > | Z| 0.3006 Median One Way Analysis Chi Square 1.0715 DF 1 Pr > Chi Square 0.3006 Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 22 of 31 AVERAGE WEIGHT OF MALE AND FEMALE FROGS AT REFERENCE AND EXPERIMENTAL SITES 138 Obs SITE SEX _ TYPE_ _ FREQ_ WEIGHT SD_ L 1 C F 0 3 36.4755 4.3213 2 C M 0 3 25.8192 9.9288 3 E F 0 5 42.8215 15.7066 4 E M 0 5 27.0119 4.1411 MEAN TOTAL WEIGHT OF TESTES ( LEFT AND RIGHT COMBINED) BY POND 139 Obs POND _ TYPE_ _ FREQ_ WEIGHT SD_ W 1 E1 0 8 0.036925 0.014242 2 E3 0 10 0.053570 0.017665 3 E4 0 10 0.057670 0.009618 4 E6 0 10 0.033560 0.018074 5 E8 0 6 0.044200 0.019885 6 R1 0 29 0.022082 0.008848 7 R3 0 17 0.030953 0.023958 8 R6 0 30 0.051621 0.013727 MEAN TOTAL WEIGHT OF TESTES ( LEFT AND RIGHT COMBINED) BY SITE 140 Obs SITE _ TYPE_ _ FREQ_ WEIGHT SD_ W 1 C 0 76 0.037232 0.021104 2 E 0 44 0.045650 0.018130 Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 23 of 31 ANOVA FOR TOTAL TESTES WEIGHT BETWEEN REFERENCE AND EXPERIMENTAL SITES 141 The ANOVA Procedure Class Level Information Class Levels Values SITE 2 C E Number of observations 120 NOTE: Due to missing values, only 91 observations can be used in this analysis.

Dependent Variable: T_ WT Sum of Source DF Squares Mean Square F Value Pr > F Model 1 0.00161041 0.00161041 4.14 0.0449 Error 89 0.03462027 0.00038899 Corrected Total 90 0.03623068 R Square Coeff Var Root MSE T_ WT Mean 0.044449 47.75261 0.019723 0.041302 Source DF Anova SS Mean Square F Value Pr > F SITE 1 0.00161041 0.00161041 4.14 0.0449 Levene's Test for Homogeneity of T_ WT Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F SITE 1 2.989E 7 2.989E 7 1.81 0.1823 Error 89 0.000015 1.654E 7 Bartlett's Test for Homogeneity of T_ WT Variance Source DF Chi Square Pr > ChiSq SITE 1 1.0070 0.3156 Dunnett's t Tests for T_ 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 89 Error Mean Square 0.000389 Critical Value of Dunnett's t 1.98699 Comparisons significant at the 0.05 level are indicated by ***.

Difference SITE Between Simultaneous 95% Comparison Means Confidence Limits E  C 0.008418 0.000197 0.016639 *** Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 24 of 31 NONPARAMETRIC COMPARISON OF TOTAL TESTES WEIGHT BETWEEN REFERENCE AND EXPERIMENTAL SITES 145 The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable T_ WT Classified by Variable SITE Sum of Expected Std Dev Mean SITE N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ C 47 1900.0 2162.0 125.912307 40.425532 E 44 2286.0 2024.0 125.912307 51.954545 Average scores were used for ties.

Wilcoxon Two Sample Test Statistic 2286.0000 Normal Approximation Z 2.0768 One Sided Pr > Z 0.0189 Two Sided Pr > | Z| 0.0378 t Approximation One Sided Pr > Z 0.0203 Two Sided Pr > | Z| 0.0407 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 4.3298 DF 1 Pr > Chi Square 0.0375 Median Scores ( Number of Points Above Median) for Variable T_ WT Classified by Variable SITE Sum of Expected Std Dev Mean SITE N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ C 47 19.0 23.241758 2.396612 0.404255 E 44 26.0 21.758242 2.396612 0.590909 Average scores were used for ties.

Median Two Sample Test Statistic 26.0000 Z 1.7699 One Sided Pr > Z 0.0384 Two Sided Pr > | Z| 0.0767 Median One Way Analysis Chi Square 3.1325 DF 1 Pr > Chi Square 0.0767 Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 25 of 31 MEAN FEMALE CONDITION INDEX BY CITE 147 Obs SITE _ TYPE_ _ FREQ_ CI SD_ CI 1 C 0 61 0.47898 0.15083 2 E 0 51 0.54278 0.19533 ANOVA FOR FEMALE CONDITION INDEX BETWEEN REFERENCE AND EXPERIMENTAL SITE 148 The ANOVA Procedure Class Level Information Class Levels Values SITE 2 C E Number of observations 112 NOTE: Due to missing values, only 110 observations can be used in this analysis.

Sum of Source DF Squares Mean Square F Value Pr > F Model 1 0.11133162 0.11133162 3.73 0.0562 Error 108 3.22722541 0.02988172 Corrected Total 109 3.33855703 R Square Coeff Var Root MSE CI Mean 0.033347 33.99081 0.172863 0.508559 Source DF Anova SS Mean Square F Value Pr > F SITE 1 0.11133162 0.11133162 3.73 0.0562 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 0.00619 0.00619 3.05 0.0835 Error 108 0.2191 0.00203 Bartlett's Test for Homogeneity of CI Variance Source DF Chi Square Pr > ChiSq SITE 1 3.5615 0.0591 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 108 Error Mean Square 0.029882 Critical Value of Dunnett's t 1.98218 Comparisons significant at the 0.05 level are indicated by ***.

Difference SITE Between Simultaneous 95% Comparison Means Confidence Limits E  C 0.06380  0.00172 0.12931 Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 26 of 31 NONPARAMETRIC COMPARISON OF FEMALE CONDITION FACTOR BETWEEN REFERENCE AND EXPERIMENTAL SIT 152 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 C 59 2965.0 3274.50 166.833000 50.254237 E 51 3140.0 2830.50 166.833000 61.568627 Wilcoxon Two Sample Test Statistic 3140.0000 Normal Approximation Z 1.8522 One Sided Pr > Z 0.0320 Two Sided Pr > | Z| 0.0640 t Approximation One Sided Pr > Z 0.0334 Two Sided Pr > | Z| 0.0667 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 3.4416 DF 1 Pr > Chi Square 0.0636 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 C 59 24.0 29.50 2.627047 0.406780 E 51 31.0 25.50 2.627047 0.607843 Median Two Sample Test Statistic 31.0000 Z 2.0936 One Sided Pr > Z 0.0181 Two Sided Pr > | Z| 0.0363 Median One Way Analysis Chi Square 4.3832 DF 1 Pr > Chi Square 0.0363 Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 27 of 31 MEAN MALE CONDITION INDEX BY SITE 154 Obs SITE _ TYPE_ _ FREQ_ CI SD_ CI 1 C 0 57 0.39435 0.13640 2 E 0 44 0.42482 0.11530 ANOVA FOR MALE CONDITION INDEX BETWEEN REFERENCE AND EXPERIMENTAL SITE 155 The ANOVA Procedure Class Level Information Class Levels Values SITE 2 C E Number of observations 101 Dependent Variable: CI Sum of Source DF Squares Mean Square F Value Pr > F Model 1 0.02304412 0.02304412 1.41 0.2372 Error 99 1.61346081 0.01629758 Corrected Total 100 1.63650493 R Square Coeff Var Root MSE CI Mean 0.014081 31.31858 0.127662 0.407624 Source DF Anova SS Mean Square F Value Pr > F SITE 1 0.02304412 0.02304412 1.41 0.2372 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 0.000694 0.000694 1.37 0.2444 Error 99 0.0501 0.000506 Bartlett's Test for Homogeneity of CI Variance Source DF Chi Square Pr > ChiSq SITE 1 1.3333 0.2482 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 99 Error Mean Square 0.016298 Critical Value of Dunnett's t 1.98422 Comparisons significant at the 0.05 level are indicated by ***.

Difference SITE Between Simultaneous 95% Comparison Means Confidence Limits E  C 0.03046  0.02037 0.08130 Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 28 of 31 NONPARAMETRIC COMPARISON OF MALE CONDITION INDEX BETWEEN REFERENCE AND EXPERIMENTAL SITES 159 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 C 57 2680.0 2907.0 146.006849 47.017544 E 44 2471.0 2244.0 146.006849 56.159091 Wilcoxon Two Sample Test Statistic 2471.0000 Normal Approximation Z 1.5513 One Sided Pr > Z 0.0604 Two Sided Pr > | Z| 0.1208 t Approximation One Sided Pr > Z 0.0620 Two Sided Pr > | Z| 0.1240 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 2.4172 DF 1 Pr > Chi Square 0.1200 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 C 57 23.0 28.217822 2.503874 0.403509 E 44 27.0 21.782178 2.503874 0.613636 Median Two Sample Test Statistic 27.0000 Z 2.0839 One Sided Pr > Z 0.0186 Two Sided Pr > | Z| 0.0372 Median One Way Analysis Chi Square 4.3426 DF 1 Pr > Chi Square 0.0372 Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 29 of 31 MEAN MALE GONADOSOMATIC INDEX ( GSI) BY SITE 161 Obs SITE _ TYPE_ _ FREQ_ GSI SD_ GSI 1 C 0 76 0.13027 0.037421 2 E 0 44 0.16847 0.042300 ANOVA FOR MALE GONADOSOMATIC ( GSI) BETWEEN REFERENCE AND EXPERIMENTAL SITE 162 The ANOVA Procedure Class Level Information Class Levels Values SITE 2 C E Number of observations 120 NOTE: Due to missing values, only 91 observations can be used in this analysis.

Dependent Variable: GSI Sum of Source DF Squares Mean Square F Value Pr > F Model 1 0.03315187 0.03315187 20.87 <. 0001 Error 89 0.14135759 0.00158829 Corrected Total 90 0.17450946 R Square Coeff Var Root MSE GSI Mean 0.189972 26.79357 0.039853 0.148742 Source DF Anova SS Mean Square F Value Pr > F SITE 1 0.03315187 0.03315187 20.87 <. 0001 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 3.249E 6 3.249E 6 0.72 0.3994 Error 89 0.000403 4.531E 6 Bartlett's Test for Homogeneity of GSI Variance Source DF Chi Square Pr > ChiSq SITE 1 0.6603 0.4164 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 89 Error Mean Square 0.001588 Critical Value of Dunnett's t 1.98699 Comparisons significant at the 0.05 level are indicated by ***. Difference SITE Between Simultaneous 95% Comparison Means Confidence Limits E  C 0.038194 0.021583 0.054806 *** Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 30 of 31 NONPARAMETRIC COMPARISON OF MALE GONADOSOMATIC INDEX BETWEEN REFERENCE AND EXPERIMENTAL SIT 166 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 C 47 1645.0 2162.0 125.915315 35.000 E 44 2541.0 2024.0 125.915315 57.750 Wilcoxon Two Sample Test Statistic 2541.0000 Normal Approximation Z 4.1020 One Sided Pr > Z <. 0001 Two Sided Pr > | Z| <. 0001 t Approximation One Sided Pr > Z <. 0001 Two Sided Pr > | Z| <. 0001 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 16.8587 DF 1 Pr > Chi Square <. 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 C 47 14.0 23.241758 2.396612 0.297872 E 44 31.0 21.758242 2.396612 0.704545 Median Two Sample Test Statistic 31.0000 Z 3.8562 One Sided Pr > Z <. 0001 Two Sided Pr > | Z| 0.0001 Median One Way Analysis Chi Square 14.8701 DF 1 Pr > Chi Square 0.0001 Data Evaluation Report on Gonadal and Laryngeal Responses to Field Exposure of Xenopus laevis to Atrazine in Areas of Corn Production in South Africa EPA MRID Number 458677 10 Page 31 of 31 MEAN CELL TYPES FOR EXPERIMENTAL SITES 168 Obs POND _ TYPE_ _ FREQ_ total Gonia Sperma Sperm other BV_ avg 1 E1 0 8 33.725 3.425 28.7625 53.35 13.675 0.75 2 E3 0 10 33.340 3.640 24.8800 58.06 13.220 0.22 3 E4 0 10 32.500 2.970 24.4900 59.82 12.580 0.16 4 E6 0 10 33.960 3.240 20.7900 62.41 13.240 0.31 5 E8 0 5 33.740 2.880 30.1400 52.24 14.540 0.16 MEAN CELL TYPES FOR REFERENCE SITES 169 Obs POND _ TYPE_ _ FREQ_ total Gonia Sperma Sperm other BV_ avg 1 C1 0 19 34.9158 3.02105 25.5737 55.9316 14.9474 0.50526 2 C3 0 17 34.3941 4.53529 34.2882 46.2647 14.6353 0.28824 3 C6 0 20 34.3800 2.82000 22.6050 60.3200 13.9050 0.33500 AVERAGE CELL TYPES BY SITE 170 Obs SITE _ TYPE_ _ FREQ_ total Gonia Sperma Sperm other BV_ avg 1 C 0 3 34.5633 3.45878 27.4890 54.1721 14.4959 0.37617 2 E 0 5 33.4530 3.23100 25.8125 57.1760 13.4510 0.32000


Comment Info: =================

General Comment:I smoked cigarettes for over 5 years, a pack a day, since I was 15 years old. The only thing that has significantly helped me to quit smoking is vaping. Hands down, it has saved my life. I would be smoking right now with no wanting to quit without vaping. I feel healthier, I feel stronger, and I don't smell absolutely terrible all the time. Even if vaping proves to be bad from a health perspective, which I doubt, I'd still do it because there is no way that it's worse than smoking cigarettes.

A 75 day comment period is insufficient time for the vaping industry and the wider vaping community to comment on the proposed deeming regulations. I would respectfully request that the period for comment be extended to 180 days.


[Federal Register Volume 87, Number 80 (Tuesday, April 26, 2022)]
[Notices]
[Pages 24551-24552]
[FR Doc No: 2022-08863]

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DEPARTMENT OF ENERGY

Federal Energy Regulatory Commission

[Docket No. AD22-4-000]

Improving Winter-Readiness of Generating Units; Third
Supplemental Notice of Technical Conference

    As announced in the Notices of Technical Conference issued in this
proceeding on November 18, 2021 and March 10, 2022, the Federal Energy
Regulatory Commission (Commission) will convene a Joint Technical
Conference with the North American Electric Reliability Corporation
(NERC) and the Regional Entities in the above-referenced proceeding on
Wednesday, April 27 and Thursday, April 28, 2022 from approximately
11:30 a.m. to 5:30 p.m. Eastern time each day. The conference will be
held virtually via WebEx.
    The purpose of this conference is to discuss how to improve the
winter-readiness of generating units, including best practices, lessons
learned, and increased use of the NERC Guidelines, as recommended in
the Joint February 2021 Cold Weather Outages Report.\1\
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    \1\ See The February 2021 Cold Weather Outages in Texas and the
South Central United States--FERC, NERC and Regional Entity Staff
Report at pp 18, 192 (November 16, 2021), https://www.ferc.gov/news-events/news/final-report-february-2021-freeze-underscores-winterization-recommendations.
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    The conference will be open for the public to attend
electronically.

Registration for the conference is not required and there is no fee for
attendance. To join the conference, go to the web Calendar of Events
for this event on FERC's website, www.ferc.gov. The link for the event
will be posted at the top of the calendar page and will ``go live''
just prior to the conference start time. The conference will also be
transcribed. Transcripts will be available for a fee from Ace
Reporting,<PHONE_NUMBER>.
    Commission conferences are accessible under section 508 of the
Rehabilitation Act of 1973. For accessibility accommodations, please
send an email to [email protected], call toll-free<PHONE_NUMBER>
(voice) or<PHONE_NUMBER> (TTY), or send a fax to<PHONE_NUMBER> with
the required accommodations.
    For more information about this technical conference, please
contact Lodie White at [email protected] or<PHONE_NUMBER>. For
information related to logistics, please contact Sarah McKinley at
[email protected] or<PHONE_NUMBER>.

    Dated: April 20, 2022.
Debbie-Anne A. Reese,
Deputy Secretary.
[FR Doc. 2022-08863 Filed 4-25-22; 8:45 am]
BILLING CODE 6717-01-P


[Federal Register: February 8, 2008 (Volume 73, Number 27)]
[Notices]
[Page 7617-7619]
[DOCID:fr08fe08-105]

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SECURITIES AND EXCHANGE COMMISSION

[Release No. 34-57260; File No. SR-BSE-2008-06]

Self-Regulatory Organizations; Boston Stock Exchange, Inc.;
Notice of Filing and Immediate Effectiveness of Proposed Rule Change
Regarding a Proposed Combination Between the Montr[eacute]al Exchange
Inc. and TSX Group Inc.

February 1, 2008
    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 January 29, 2008, the Boston Stock Exchange, Inc. (``BSE'' 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 prepared by BSE. The Exchange filed the
proposal as a ``non-controversial'' proposed rule change pursuant to
Section 19(b)(3)(A)(iii) 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.
---------------------------------------------------------------------------

    \1\ 15 U.S.C. 78s(b)(1).
    \2\ 17 CFR 240.19b-4.
    \3\ 15 U.S.C. 78s(b)(3)(A)(iii).
    \4\ 17 CFR 240.19b-4(f)(6).
---------------------------------------------------------------------------

I. Self-Regulatory Organization's Statement of the Terms of Substance
of the Proposed Rule Change

    The Exchange is submitting the proposed rule change to the
Commission to amend the Fifth Amended and Restated Operating Agreement,
dated January 26, 2005, as may be amended from time to time (``BOX LLC
Agreement''), of the Boston Options Exchange Group LLC (``BOX LLC''),
in connection with the proposed business combination (the
``Combination'') of the Montr[eacute]al Exchange Inc.,\5\ a company
incorporated in Qu[eacute]bec, Canada (``MX''), and TSX Group Inc., a
company incorporated in Ontario, Canada (``TSX Group''). The text of
the rule proposal, including the proposed Instrument of Accession, is
available on the Exchange's Web site (http://www.bostonstock.com), at

the Exchange, and at the Commission's Public Reference Room.
---------------------------------------------------------------------------

    \5\ The Montr[eacute]al Exchange Inc. is also known in French as
the Bourse de Montr[eacute]al Inc.
---------------------------------------------------------------------------

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

    In its filing with the Commission, BSE 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. BSE 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
Statutory Basis for, the Proposed Rule Change

1. Purpose
    On January 13, 2004, the Commission approved four BSE proposals
that together established, through an operating agreement among its
owners, a Delaware limited liability company, BOX LLC, to operate BOX
as an options trading facility of the Exchange.\6\ Currently, MX U.S.
2, Inc., a wholly owned U.S. subsidiary of MX (``MX US''), owns a 31.4%
ownership interest in BOX LLC.
---------------------------------------------------------------------------

    \6\ See Securities Exchange Act Release Nos. 49066 (January 13,
2004), 69 FR 2773 (January 20, 2004) (establishing a fee schedule
for the proposed BOX facility); 49065 (January 13, 2004), 69 FR 2768
(January 20, 2004) (creating Boston Options Exchange Regulation LLC
to which the BSE would delegate its self-regulatory functions with
respect to the BOX facility); 49068 (January 13, 2004), 69 FR 2775
(January 20, 2004) (approving trading rules for the BOX facility);
and 49067 (January 13, 2004), 69 FR 2761 (January 20, 2004)
(approving certain regulatory provisions of the operating agreement
of BOX LLC).
---------------------------------------------------------------------------

    The Exchange is submitting the proposed rule change to the
Commission to amend the BOX LLC Agreement pursuant to the proposed
Instrument of Accession in connection with the Combination of MX, the
largest derivatives exchange in Canada, and TSX Group, which, among
other things, owns Canada's pre-eminent equity market. As a result of
the Combination, MX will become a direct \7\ subsidiary of TSX Group, a
publicly traded Ontario corporation.\8\
    The Combination will be effected through a series of amalgamations
\9\ involving intermediate holding companies, which will cease to exist
following completion of the Combination. As a result of these
transactions, MX will be amalgamated with a direct subsidiary of TSX
Group. The amalgamated company also will be named Montr[eacute]al
Exchange Inc. and will be a Qu[eacute]bec corporation. Consequently, MX
U.S. (including MX US's 31.4% ownership interest in BOX LLC) will
become an indirect, wholly owned subsidiary of TSX Group. The
Combination is subject to the approval of the Autorit[eacute] des
march[eacute]s financiers (the securities regulator for the province of
Qu[eacute]bec, Canada) and the shareholders of MX, as well as other
regulatory approvals.
---------------------------------------------------------------------------

    \7\ Certain non-voting preference shares of MX will be owned by
TSX Inc., a direct, wholly owned subsidiary of TSX Group. The share
interests of TSX Group and TSX Inc. will together represent the
entire ownership interest, voting and non-voting, in MX.
    \8\ At its next shareholders' meeting after the effective date
of the Combination, TSX Group will propose changing its name to TMX
Group Inc.
    \9\ The term ``amalgamation'' refers to the combination of two
or more business entities into a single entity pursuant to which the
combined entity becomes the successor in interest, by operation of
law, to the rights and obligations of the combining entities.
Amalgamations are commonly used in Canada to effect business
combinations and are similar to mergers in the United States.
---------------------------------------------------------------------------

    Following the Combination, MX, MX US, and TSX Group will continue
to operate their respective businesses in substantially the same manner
as they had prior to the Combination, and MX and MX US senior
management will remain under the stewardship of MX's current chief
executive officer.\10\ Additionally, the operations of each of MX and
TSX Group will continue to be located in the same province in which it
is currently located, and each will remain subject to its existing
regulatory framework and oversight. Consequently, MX US's management of
its ownership interest in BOX shall remain essentially unaffected by
the Combination.
---------------------------------------------------------------------------

    \10\ Organizational changes contemplated in connection with the
Combination are described in the TSX Group registration statement
separately filed with the Commission regarding the issuance of
shares in connection with the Combination.
---------------------------------------------------------------------------

    Pursuant to Section 8.4(g) of the BOX LLC Agreement, BOX LLC is
required to amend the BOX LLC Agreement to make an Acquirer \11\ a
party to the LLC Agreement if such Acquirer acquires a Controlling
Interest \12\ in a BOX Member who holds a percentage interest in BOX
LLC equal to or greater than 20%. Therefore, since TSX Group is
acquiring a Controlling Interest in MX, whose wholly owned subsidiary,
MX US, owns a 31.4% ownership interest in BOX LLC, TSX Group, as an
Acquirer, is required to and will become a party to the BOX LLC
Agreement pursuant to the proposed Instrument of Accession. As a
result, TSX Group will agree to abide by all the provisions of the BOX
LLC Agreement, including those provisions requiring submission to the
jurisdiction of the Commission.\13\
---------------------------------------------------------------------------

    \11\ An ``Acquirer'' is defined as ``a Person who, alone or
together with any Affiliate of such Person, acquires a controlling
interest in a Member.'' See BOX LLC Agreement, Section 8.4(g)(ii).
    \12\ A ``Controlling Interest'' is defined as ``the ownership by
any Person, alone or together with any Affiliate of such Person, of
a 25% or greater interest in a Member.'' See BOX LLC Agreement,
Section 8.4(g)(i).
    \13\ The BOX LLC Agreement states, in part, that ``the Members,
officers, directors, agents, and employees of Members irrevocably
submit to the exclusive jurisdiction of the U.S. federal courts,
U.S. Securities and Exchange Commission, and the Boston Stock
Exchange, for the purposes of any suit, action or proceeding
pursuant to U.S. federal securities laws, the rules or regulations
thereunder, arising out of, or relating to, BOX activities or
Article 19.6(a), (except that such jurisdictions shall also include
Delaware for any such matter relating to the organization or
internal affairs of BOX, provided that such matter is not related to
trading on, or the regulation, of the BOX Market), and hereby waive,
and agree not to assert by way of motion, as a defense or otherwise
in any such suit, action or proceeding, any claims that they are not
personally subject to the jurisdiction of the U.S. Securities and
Exchange Commission, that the suit, action or proceeding is an
inconvenient forum or that the venue of the suit, action or
proceeding is improper, or that the subject matter hereof may not be
enforced in or by such courts or agency.'' See BOX LLC Agreement,
Section 19.6.
---------------------------------------------------------------------------

    For the reasons stated above, BSE is submitting to the Commission
the proposed Instrument of Accession, which constitutes an amendment to
the BOX LLC Agreement, as a rule change. The proposed rule change is
subject to becoming effective and operative pursuant to Section
19(b)(3)(A) of the Act \14\ and Rule 19b-4(f)(6) \15\ thereunder. BSE
proposes to make this proposal operative upon the consummation of the
Combination, currently anticipated in late February 2008.
---------------------------------------------------------------------------

    \14\ 15 U.S.C. 78s(b)(3)(A).
    \15\ 17 CFR 240.19b-4(f)(6).
---------------------------------------------------------------------------

2. Statutory Basis
    The Exchange believes that the proposal is consistent with the
requirements of Section 6(b) of the Act,\16\ in general, and furthers
the objectives of Section 6(b)(1),\17\ in particular, in that it
enables the Exchange to be so organized so as to have the capacity to
be able to carry out the purposes of the Act and to comply, and to
enforce compliance by its exchange members and persons associated with
its exchange members, with the provisions of the Act, the rules and
regulations thereunder, and the rules of the Exchange.
---------------------------------------------------------------------------

    \16\ 15 U.S.C. 78f(b).
    \17\ 15 U.S.C. 78f(b)(1).
---------------------------------------------------------------------------

    The Exchange also believes that this filing furthers the objectives
of Section 6(b)(5) of the Act \18\ in that it is designed to facilitate
transactions in securities, to prevent fraudulent and manipulative acts
and practices, to promote just and equitable principles of trade, to
foster cooperation and coordination with persons engaged in regulating,
clearing, settling, processing information with respect to, and
facilitating 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.
---------------------------------------------------------------------------

    \18\ 15 U.S.C. 78f(b)(5).
---------------------------------------------------------------------------

    Additionally, the Exchange notes that the provisions of the BOX LLC
Agreement, previously approved by the Commission, provide a framework
for addressing the Combination. Accordingly, BSE believes the
Combination does not present any novel

issues that have not been anticipated and addressed by the BOX LLC
Agreement.

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 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

    The Exchange has neither solicited nor received comments on 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 \19\ and Rule 19b-
4(f)(6) thereunder.\20\
---------------------------------------------------------------------------

    \19\ 15 U.S.C. 78s(b)(3)(A).
    \20\ 17 CFR 240.19b-4(f)(6). In addition, Rule 19b-4(f)(6)(iii)
requires that a self-regulatory organization submit to the
Commission 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 filing of
the proposed rule change, or such shorter time as designated by the
Commission. The Commission notes that BSE has satisfied the five-day
pre-filing notice requirement.
---------------------------------------------------------------------------

    Normally, a proposed rule change filed under Rule 19b-4(f)(6) \21\
may not become operative prior to 30 days after the date of filing.
However, Rule 19b-4(f)(6)(iii) \22\ 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. In its filing, the
Exchange requested waiver of the 30-day operative delay because the MX
shareholders are expected to approve the Combination on February 13,
2008 and subsequent thereto the Combination is expected to close in
late February of 2008. Furthermore, BSE believes the Combination does
not present any novel issues that have not been anticipated and
addressed by the BOX LLC Agreement.
---------------------------------------------------------------------------

    \21\ 17 CFR 240.19b-4(f)(6).
    \22\ 17 CFR 240.19b-4(f)(6)(iii).
---------------------------------------------------------------------------

    The Commission believes that waiving the 30-day operative delay is
consistent with the protection of investors and the public interest
because the proposed rule change will allow the Exchange to proceed
with the Combination, without undue delay, in a manner consistent with
the provisions of the BOX LLC Operating Agreement. Accordingly,
consistent with the protection of investors and the public interest,
the Commission designates the proposed rule change to be operative upon
consummation of the Combination.\23\
---------------------------------------------------------------------------

    \23\ For the purposes only of waiving the 30-day operative
delay, the Commission has considered the proposed rule's impact on
efficiency, competition, and capital formation. See 15 U.S.C.
78c(f).
---------------------------------------------------------------------------

    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-BSE-2008-06 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-BSE-2008-06. 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 BSE. 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-BSE-2008-06 and should be
submitted on or before February 29, 2008.

    For the Commission, by the Division of Trading and Markets,
pursuant to delegated authority.\24\
---------------------------------------------------------------------------

    \24\ 17 CFR 200.30-3(a)(12).
---------------------------------------------------------------------------

Florence E. Harmon,
Deputy Secretary.
 [FR Doc. E8-2329 Filed 2-7-08; 8:45 am]

BILLING CODE 8011-01-P


[Federal Register Volume 79, Number 70 (Friday, April 11, 2014)]
[Notices]
[Pages 20285-20287]
[FR Doc No: 2014-08123]

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SECURITIES AND EXCHANGE COMMISSION

[Release No. 34-71888; File No. SR-DTC-2014-02]

Self-Regulatory Organizations; The Depository Trust Company;
Notice of Filing and Immediate Effectiveness of Proposed Rule Change to
Amend the DTC Settlement Service Guide to Clarify the Largest
Provisional Net Credit Procedures

April 7, 2014.
    Pursuant to Section 19(b)(1) of the Securities Exchange Act of 1934

(``Act'') \1\ and Rule 19b-4 \2\ thereunder, notice is hereby given
that on March 27, 2014, the Depository Trust Company (``DTC'') filed
with the Securities and Exchange Commission (``Commission'') the
proposed rule change \3\ as described in Items I, II and III below,
which Items have been prepared primarily by DTC. DTC filed the proposed
rule change pursuant to Section 19(b)(3)(A) \4\ of the Act and Rule
19b-4(f)(4) \5\ thereunder, 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.
---------------------------------------------------------------------------

    \1\ 15 U.S.C. 78s(b)(1).
    \2\ 17 CFR 240.19b-4.
    \3\ Exhibit 5 of this proposed rule change filing shows the text
of the rule changes to be made by DTC. File No. SR-DTC-2014-02,
Exhibit 5, http://www.sec.gov/rules/sro/dtc.shtml.
    \4\ 15 U.S.C. 78s(b)(3)(A).
    \5\ 17 CFR 240.19b-4(f)(4).
---------------------------------------------------------------------------

I. Self-Regulatory Organization's Statement of the Terms of Substance
of the Proposed Rule Change

    As discussed below, the proposed rule change would clarify DTC's
existing Largest Provisional Net Credit (``LPNC'') \6\ Procedures
within DTC's Settlement Service Guide (the ``Service Guide'').\7\
---------------------------------------------------------------------------

    \6\ DTC Rule 1 (Definitions), available at http://www.dtcc.com/en/legal/rules-and-procedures.aspx.
    \7\ Service Guide, available at http://www.dtcc.com/~/media/
Files/Downloads/legal/service-guides/Settlement.ashx.
---------------------------------------------------------------------------

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

    In its filing with the Commission, DTC 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. DTC 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
Statutory Basis for, the Proposed Rule Change

1. Purpose
    With regard to processing of Money Market Instruments (``MMI''),
DTC employs the LPNC risk management control to mitigate the risks
associated, on the same business day, with: (i) the insolvency of an
MMI issuer or refusal to pay (``RTP'') on a MMI maturity obligation by
the applicable MMI Issuing and Paying Agent, and (ii) a DTC
Participant's (``Participant'') failure to settle or DTC's ceasing to
act for the Participant. In this regard, on each processing day, DTC
withholds intraday credit from each MMI Participant for the sum of the
two largest aggregate net credits with respect to an issuer's acronym
for purposes of calculating the Participant's net settlement balance
and collateral monitor. This provides protection in the event that MMI
transactions are reversed in DTC's system in accordance with the DTC
Rules & Procedures (``Rules'') as a result of an issuer failure or
RTP.\8\
---------------------------------------------------------------------------

    \8\ Such reversals override DTC's net debit cap and collateral
monitor risk controls.
---------------------------------------------------------------------------

    DTC most recently updated its Rules relating to LPNC in 2013.\9\
---------------------------------------------------------------------------

    \9\ Securities Exchange Act Release No. 68983 (Feb. 25, 2013),
78 FR 13924 (Mar. 1, 2013) (SR-DTC-2012-10).
---------------------------------------------------------------------------

    Pursuant to this rule filing, DTC proposes clarifications to the
LPNC Procedures in the Service Guide, including: (i) providing an
example illustrating the composition of the LPNC calculation; and (ii)
making other technical changes relating to the description of the LPNC
process and timeframes.
2. Statutory Basis
    The proposed rule change clarifies the terms associated with
processing of MMI transactions and associated risk controls. As a
result, the proposed rule change is consistent with the requirements of
the Act and the rules and regulations thereunder applicable to DTC, and
in particular, with Section 17A(b)(3)(F) \10\ of the Act. This section
requires that the Rules be designed to promote the prompt and accurate
clearance and settlement of securities transactions and, in general, to
protect investors and the public interest.
---------------------------------------------------------------------------

    \10\ 15 U.S.C. 78q-1(b)(3)(F).
---------------------------------------------------------------------------

3. Implementation Timeframe
    The proposed rule change is effective immediately.

(B) Self-Regulatory Organization's Statement on Burden on Competition

    DTC does not believe that the proposed rule change will have any
impact, or impose any burden, on competition. As stated above, the
proposed change merely provides clarifications to the Service Guide and
will not impact any Participant's access to, or use of, DTC services.
In addition, the proposal does not impact the costs of Participants'
use of DTC services.

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

    Written comments relating to the proposed rule change have not yet
been solicited or received with respect to this filing.

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

    The forgoing [sic] rule change has become effective pursuant to
Section 19(b)(3)(A) \11\ of the Act and paragraph (f)(4) \12\ of Rule
19b-4 thereunder. At any time within 60 days of the filing of the
proposed rule change, the Commission summarily may temporarily suspend
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.
---------------------------------------------------------------------------

    \11\ 15 U.S.C. 78s(b)(3)(A).
    \12\ 17 CFR 240.19b-4(f)(4).
---------------------------------------------------------------------------

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 email to<EMAIL_ADDRESS>Please include
File No. SR-DTC-2014-02 on the subject line.

Paper Comments

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

All submissions should refer to File No. SR-DTC-2014-02. This file
number should be included on the subject line if email 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 Web site viewing and
printing in the Commission's Public Reference Room, 100 F Street NE.,
Washington, DC 20549 on official business days between the hours of
10:00 a.m. and 3:00 p.m. Copies of such filings will also be available
for inspection and copying at the principal office of DTC and on DTC's
Web site at http://www.dtcc.com/legal/sec-rule-filings.aspx. 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 No. SR-DTC-2014-02 and should be
submitted on or before May 2, 2014.

    For the Commission, by the Division of Trading and Markets,
pursuant to delegated authority.\13\
Kevin M. O'Neill,
Deputy Secretary.
---------------------------------------------------------------------------

    \13\ 17 CFR 200.30-3(a)(12).
---------------------------------------------------------------------------

[FR Doc. 2014-08123 Filed 4-10-14; 8:45 am]
BILLING CODE 8011-01-P


[Federal Register Volume 83, Number 61 (Thursday, March 29, 2018)]
[Proposed Rules]
[Pages 13440-13442]
[FR Doc No: 2018-06251]

=======================================================================
-----------------------------------------------------------------------

DEPARTMENT OF HEALTH AND HUMAN SERVICES

Food and Drug Administration

21 CFR Chapter I

[Docket No. FDA-2018-N-1214]

Medical Gas Regulation; Public Workshop; Request for Comments

AGENCY: Food and Drug Administration, HHS.

ACTION: Request for comments; public workshop.

-----------------------------------------------------------------------

SUMMARY: The Food and Drug Administration (FDA, the Agency, or we) is
announcing an additional public workshop on medical gas regulation
entitled ``Medical Gas Regulation: Workshop III.'' FDA has previously
held two public workshops entitled ``Medical Gas Regulation: Workshop
I'' and ``Medical Gas Regulation: Workshop II.'' The topic to be
discussed is potential areas of Federal drug regulation that should be
revised with respect to medical gases.

DATES: The public workshop will be held on May 11, 2018, from 9 a.m. to
5 p.m. Submit either electronic or written comments on this public
workshop by August 9, 2018. See the SUPPLEMENTARY INFORMATION section
for registration date and information.

ADDRESSES: The public workshop will be held at FDA's White Oak Campus,
10903 New Hampshire Ave., Bldg. 31 Conference Center, Rm. 1503 (the
``Great Room''), Silver Spring, MD 20993-0002. Entrance for public
workshop participants (non-FDA employees) is through Building 1 where
routine security check procedures will be performed. For parking and
security information, please refer to https://www.fda.gov/AboutFDA/WorkingatFDA/BuildingsandFacilities/WhiteOakCampusInformation/ucm241740.htm. The workshop is free and seating will be on a first-
come, first-served basis. Attendees who do not wish to make an oral
presentation do not need to register.
    You may submit comments as follows. Please note that late, untimely
filed comments may not be considered. For timely consideration, we
request that electronic comments on workshop topics be submitted before
or within 90 days after each workshop (i.e., comments submitted by or
before March 15, 2018, for Workshop I; May 10, 2018, for Workshop II;
and August 9, 2018, for Workshop III). FDA has one shared docket for
all workshops. However, with this notice, the docket number will change
from FDA-2017-N-0001 to FDA-2018-N-1214. All comments submitted on the
previous docket number will be transferred to the new docket number.
The https://www.regulations.gov electronic filing system will accept
comments until midnight Eastern Time at the end of August 9, 2018.
Comments received by mail/hand delivery/courier (for written/paper
submissions) will be considered timely if they are postmarked or the
delivery service acceptance receipt is on or before the relevant date.

Electronic Submissions

    Submit electronic comments in the following way:
     Federal eRulemaking Portal: https://www.regulations.gov.
Follow the instructions for submitting comments. Comments submitted
electronically, including attachments, to https://www.regulations.gov
will be posted to the docket unchanged. Because your comment will be
made public, you are solely responsible for ensuring that your comment
does not include any confidential information that you or a third party
may not wish to be posted, such as medical information, your or anyone
else's Social Security number, or confidential business information,
such as a manufacturing process. Please note that if you include your
name, contact information, or other information that identifies you in
the body of your comments, that information will be posted on https://www.regulations.gov.
     If you want to submit a comment with confidential
information that you

do not wish to be made available to the public, submit the comment as a
written/paper submission and in the manner detailed (see ``Written/
Paper Submissions'' and ``Instructions'').

Written/Paper Submissions

    Submit written/paper submissions as follows:
     Mail/Hand delivery/Courier (for written/paper
submissions): Dockets Management Staff (HFA-305), Food and Drug
Administration, 5630 Fishers Lane, Rm. 1061, Rockville, MD 20852.
     For written/paper comments submitted to the Dockets
Management Staff, FDA will post your comment, as well as any
attachments, except for information submitted, marked and identified,
as confidential, if submitted as detailed in ``Instructions.''
    Instructions: All submissions received must include the Docket No.
FDA-2018-N-1214 for ``Medical Gas Regulation.'' Received comments,
those filed in a timely manner (see ADDRESSES), will be placed in the
docket and, except for those submitted as ``Confidential Submissions,''
publicly viewable at https://www.regulations.gov or at the Dockets
Management Staff between 9 a.m. and 4 p.m., Monday through Friday.
     Confidential Submissions--To submit a comment with
confidential information that you do not wish to be made publicly
available, submit your comments only as a written/paper submission. You
should submit two copies total. One copy will include the information
you claim to be confidential with a heading or cover note that states
``THIS DOCUMENT CONTAINS CONFIDENTIAL INFORMATION.'' The Agency will
review this copy, including the claimed confidential information, in
its consideration of comments. The second copy, which will have the
claimed confidential information redacted/blacked out, will be
available for public viewing and posted on https://www.regulations.gov.
Submit both copies to the Dockets Management Staff. If you do not wish
your name and contact information to be made publicly available, you
can provide this information on the cover sheet and not in the body of
your comments and you must identify this information as
``confidential.'' Any information marked as ``confidential'' will not
be disclosed except in accordance with 21 CFR 10.20 and other
applicable disclosure law. For more information about FDA's posting of
comments to public dockets, see 80 FR 56469, September 18, 2015, or
access the information at: https://www.gpo.gov/fdsys/pkg/FR-2015-09-18/pdf/2015-23389.pdf.
    Docket: For access to the docket to read background documents or
the electronic and written/paper comments received, go to https://www.regulations.gov and insert the docket number, found in brackets in
the heading of this document, into the ``Search'' box and follow the
prompts and/or go to the Dockets Management Staff, 5630 Fishers Lane,
Rm. 1061, Rockville, MD 20852.

FOR FURTHER INFORMATION CONTACT: Christine Kirk, Center for Drug
Evaluation and Research, Food and Drug Administration, 10903 New
Hampshire Ave., Silver Spring, MD 20993-0002,<PHONE_NUMBER>, Fax: 301-
847-8440, email: [email protected].

SUPPLEMENTARY INFORMATION:

I. Background

    On May 5, 2017, President Trump signed the Consolidated
Appropriations Act of 2017 (Pub. L. 115-31). Section 756 of the
Consolidated Appropriations Act requires FDA to issue final regulations
revising Federal drug regulations with respect to medical gases. These
public workshops are being held as part of FDA's implementation of the
requirements of section 756.
    Since the enactment of the Food and Drug Administration Safety and
Innovation Act (FDASIA) (Pub. L. 112-144), FDA has engaged in multiple
activities related to medical gases, including rulemaking. For example,
in 2016, FDA issued the final rule ``Medical Gas Containers and
Closures: Current Good Manufacturing Practice Requirements'' (81 FR
81685; November 18, 2016). Other activities include FDA's June 2017
revised draft guidance for industry on current good manufacturing
practice for medical gases,\1\ updated guidance for FDA inspectors
regarding medical gases (March 2015),\2\ an extensive review of Federal
drug regulations related to medical gases from 2012 to 2014 (a report
on the review was submitted to Congress in 2015),\3\ and implementation
of FDASIA's requirements regarding certification of medical gases (to
date, over 70 certifications have been granted).
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    \1\ Available at: https://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/ucm070270.pdf.
    \2\ Available at: https://www.fda.gov/downloads/ICECI/ComplianceManuals/ComplianceProgramManual/UCM125417.pdf.
    \3\ Available at: https://www.fda.gov/downloads/regulatoryinformation/lawsenforcedbyfda/significantamendmentstothefdcact/fdasia/ucm453727.pdf.
---------------------------------------------------------------------------

    FDA intends to engage in additional rulemaking in this area in
accordance with section 756 of the Consolidated Appropriations Act of
2017. To conduct rulemaking as efficiently as possible, FDA intends to
build on the information and stakeholder input received since FDASIA's
enactment. As noted in more detail below, FDA invites comments from
stakeholders on specific medical gas issues that could or should be
addressed in regulation.

II. Topics for Discussion at the Public Workshops

    We are holding these workshops to provide an opportunity for
medical gas manufacturers and any other interested members of the
public to provide input on potential areas of Federal drug regulation
that should be revised with respect to medical gases.
    We are asking stakeholders to comment on existing medical gas
issues that, in their view, should be addressed by regulation change
(rather than through other means, such as revisions to guidance or
inspection practices). Commenters should include concrete and specific
reasons that rulemaking is preferable to other options. Commenters'
views regarding the prioritization of particular rulemaking proposals
would also be helpful. As noted above, the https://www.regulations.gov
electronic filing system will accept comments until midnight Eastern
Time at the end of August 9, 2018. Late comments may not be considered.
    During Workshop I (December 2017), FDA and workshop participants
discussed the anticipated scope of the medical gas rulemaking, as well
as three regulations to which stakeholders have previously requested
changes: Part 201 (21 CFR part 201) (labeling generally and labeling
for medical air specifically), part 207 (21 CFR part 207) (registration
and listing), and parts 210 and 211 (21 CFR parts 210 and 211) (current
good manufacturing practice). A stakeholder presentation addressed
parts 201, 210, and 211, among other things, including initial
stakeholder views on the possibility of having one or more separate CFR
sections for designated medical gases. FDA also heard comments on
additional regulations and medical gas issues as time allowed.
    During Workshop II (February 2018), FDA and workshop participants
discussed parts 310, 314, and 514 (21 CFR parts 310, 314, and 514)
(postmarket reporting of adverse drug experiences, including adverse
reactions and medication errors) and the intersection of regulations
for medical gases and regulations for medical devices and animal drugs.
A stakeholder

presentation also addressed, among other things, followup information
related to Workshop I topics, including part 207 (registration and
listing) and parts 210 and 211 (current good manufacturing practice),
including the possibility of one or more separate CFR sections for
designated medical gases, as well as additional topics including the
certification process for designated medical gases and issues related
to the filling of oxygen containers by emergency medical service (EMS)
providers and health care facilities. FDA also heard comments on
additional regulations and medical gas issues as time allowed.
    The Agency has determined that we will hold a third workshop to
hear additional comments from stakeholders regarding the issues
discussed at Workshops I and II, as well as any additional topics
related to medical gas regulation that stakeholders may wish to
discuss, as time allows. This workshop is primarily intended to build
on the discussion from the previous workshops, as well as written
comments submitted to the docket.
    During Workshop III (May 11, 2018), FDA intends to provide
designated panel time for followup discussion of several topics raised
at previous workshops, and for an open panel to discuss any additional
issues related to medical gas regulation that are of interest to FDA or
other workshop participants. The topics for designated panel time
include further consideration of potential changes to: Part 201
(labeling); parts 210 and 211 (current good manufacturing practice);
part 207 (registration and listing); and parts 310, 314, and 514
(postmarket reporting of adverse drug experiences, including adverse
reactions and medication errors); including the possibility of one or
more separate CFR sections for designated medical gases. Potential
topics for open panel time include, but are not limited to: The
certification process for designated medical gases; issues related to
the filling of oxygen containers by EMS providers and health care
facilities; or other topics of interest to stakeholders.

III. Participating in the Public Workshop

    Registration and Requests for Oral Presentations: If you wish to a
make an oral presentation, you must register by submitting your name,
title, firm name, address, telephone, email address, and Fax number to
[email protected] (see FOR FURTHER INFORMATION CONTACT)
by May 4, 2018, for Workshop III. Please also indicate the type of
organization you represent (e.g., industry, consumer organization) and
a brief summary of your remarks (including the discussion topic(s) that
you would like to address).
    FDA will try to accommodate all persons who wish to make a
presentation; however, the duration of each speaker's presentation may
be limited by time constraints. FDA will notify registered presenters
of their scheduled presentation times. Persons registered to speak
should check in before the workshop and are encouraged to arrive early
to ensure their designated order of presentation. Participants who are
not present when called may not be permitted to speak at a later time.
An agenda will be made available at least 3 days before the workshop at
https://www.fda.gov/Drugs/NewsEvents/ucm582091.htm. FDA may also post
specific questions for consideration at the meeting web page; these
will be made available at least 3 days before the workshop at https://www.fda.gov/Drugs/NewsEvents/ucm582091.htm.
    Streaming Webcast of the Public Workshops: This public workshop
will be webcast; the URL will be posted at https://www.fda.gov/Drugs/NewsEvents/ucm582091.htm at least 1 day before the workshop. A video
record of the public workshops will be available at the same website
address for 1 year. If you need special accommodations because of a
disability, please contact [email protected] (or see
FOR FURTHER INFORMATION CONTACT) at least 7 days in advance of the
workshop.

    Dated: March 21, 2018.
Leslie Kux,
Associate Commissioner for Policy.
[FR Doc. 2018-06251 Filed 3-28-18; 8:45 am]
 BILLING CODE 4164-01-P


[Federal Register Volume 79, Number 215 (Thursday, November 6, 2014)]
[Notices]
[Pages 65934-65935]
[FR Doc No: 2014-26389]

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DEPARTMENT OF ENERGY

Federal Energy Regulatory Commission

[Docket No. TS13-4-000]

Hudson Transmission Partners, LLC; Notice of Filing

    Take notice that on October 29, 2014, Hudson Transmission Partners,
LLC filed a supplement to its July 12, 2013 request for exemption from,
or waiver of, the standards of conduct set forth in Part 358 of the
Federal Energy Regulatory Commission's (Commission) regulations, 18 CFR
358.
    Any person desiring to intervene or to protest this filing must
file in accordance with Rules 211 and 214 of the Commission's Rules of
Practice and Procedure (18 CFR 385.211, 385.214). Protests will be
considered by the Commission in determining the appropriate action to
be taken, but will not serve to make protestants parties to the
proceeding. Any person wishing to become a party must file a notice of
intervention or motion to intervene, as appropriate. Such notices,
motions, or protests must be filed on or before the comment date. On or
before the comment date, it is not necessary to serve motions to
intervene or protests on persons other than the Applicant.
    The Commission encourages electronic submission of protests and
interventions in lieu of paper using the ``eFiling'' link at http://www.ferc.gov. Persons unable to file electronically should submit an
original and 5 copies of the protest or intervention to the Federal
Energy Regulatory Commission, 888 First Street NE., Washington, DC
20426.
    This filing is accessible on-line at http://www.ferc.gov, using the
``eLibrary'' link and is available for review in the Commission's
Public Reference Room in Washington, DC.

There is an ``eSubscription'' link on the Web site that enables
subscribers to receive email notification when a document is added to a
subscribed docket(s). For assistance with any FERC Online service,
please email<EMAIL_ADDRESS>or call<PHONE_NUMBER> (toll
free). For TTY, call<PHONE_NUMBER>.
    Comment Date: 5:00 p.m. Eastern Time on November 19, 2014.

    Dated: October 30, 2014.
Kimberly D. Bose,
Secretary.
[FR Doc. 2014-26389 Filed 11-5-14; 8:45 am]
BILLING CODE 6717-01-P