**UNITED ST AT**  **ES ENVIRONMENT AL**  **PROTECTION A GENCY**  **OFFICE OF CHEMICAL SAFETY AND POLLUTION PREVENTION MEMORANDUM**  February 23 , 2011 **SUBJECT: Acibenzolar**  **- S**  **- methyl** 

Acibenzolar - S - methyl **PC Code:** 061402 **Decision No.:** 443561 **Petition No.:** 7E7818 **Risk Assessment Type:** Single Chemical Aggregate **T XR No.:**  NA **MRID No.:** NA **FROM:** Becky Daiss , Biologist

James Scott Miller Thurston Morton , Abdallah Khasawinah, Toxicologist Risk Assessment Branch Health Effects Division (7509 **THROUGH:** Elissa Reaves , PhD,

Susan V. Hummel, Branch Senior Scientist Risk Assessment **TO:** Sydney

Jackson , Risk Manager Review Risk Integration, Minor Use & Emergency Response Branch Registration Division This document provides the acibenzolar - S - methyl on low growing berries, crop subgroup 13

Page 1 of 47 **UNITED ST AT**  **ES ENVIRONMENT AL**  **PROTECTION A GENCY** 

**WA SHINGTON, D.C. 20460**  **OFFICE OF CHEMICAL SAFETY AND POLLUTION PREVENTION**

**methyl** Human Health Risk Assessment for Propos methyl on Lo w Growing Berries Crop Subgroup 13 **DP Barcode:** 3 87071 **Registration No. :**  100 - RGAG **Regulatory Action:** Section 3 Registration Single Chemical Aggregate **Case No.:** 7031 **CAS No.:** 135158 - 54 - 2 **40 CFR:** 180. 561 , Biologist Miller , Environmental Scientist , Chemist Abdallah Khasawinah, Toxicologist Risk Assessment Branch 4 Health Effects Division (7509 P )

PhD, Chief Susan V. Hummel, Branch Senior Scientist

Branch 4 , HED (7509 P ) , Risk Manager Review Risk Integration, Minor Use & Emergency Response Branch Registration Division (750 5 P)

the Health Effects Division’s (HED’s) risk assessment of low growing berries, crop subgroup 13 - 07G .

Human Health Risk Assessment for Propos ed Use of Lo w Growing Berries Crop Subgroup 13 - 07G . RGAG Section 3 Registration

Health Effects Division’s (HED’s) risk assessment of use ofPage 2 of 47

TABLE OF CONTENTS pg. 1.0 EXECUTIVE SUMMARY ................................ ................................ ................................ . 4 2.0 HED RECOMMENDATIONS ................................ ................................ ............................ 6 2.1 Data Deficiencies/Conditions of Registration ................................ ................................ ..... 6 2.2 Tolerance Considerations ................................ ................................ ................................ ..... 6 2.2.1 Enforcement Anal yt ical Method ................................ ................................ .............. 6 2.2.2 International Harmonization ................................ ................................ .................... 6 2.2.3 Recommended Tolerances ................................ ................................ ....................... 6 2.2.4 Revisions to Petitioned - For Tolerances ................................ ................................ ... 7 3.0 IN GREDIENT PROFILE ................................ ................................ ................................ .... 7 3.1 Chemical Identity ................................ ................................ ................................ ................ 7 3. 2 Phy sical/Chemical Properties ................................ ................................ .............................. 8 3.3 Pesticide Use Pattern ................................ ................................ ................................ ............ 8 3.3.1 Registered Products ................................ ................................ ................................ . 8 3.3.2 Registered and Propose d New Uses ................................ ................................ ........ 8 3.4 Anticipated Exposure Pathway s ................................ ................................ .......................... 9 3.5 Consideration of Environmental Justice ................................ ................................ .............. 9 4.0 HAZARD CHARACTERIZATION/ASSESSMENT ................................ ......................... 9 4.1 Toxicology

Studies Available for Analy sis ................................ ................................ ......... 9 4.2 Absorption, Distribution, Metabolism, & Elimination ................................ ........................ 9 4.3 Toxicolo gical Effects ................................ ................................ ................................ ......... 1 0 4.4 Safety

Factor for Infants and Children ................................ ................................ ............... 1 1 4.4.1 Completeness of the Toxicology Database ................................ ............................ 12 4.4.2 Evidence of Neurotoxicity ................................ ................................ ..................... 12 4.4.3 Evidence of Sensitivity /susceptibility

in the Developing or Young ...................... 1 2 4.4.4 Residual Uncertaint y in the Exposure Data Base ................................ .................. 1 3 4.5 Toxicity

Endpoint and Point of Departure Selections ................................ ....................... 13 4.5.1 Dose - Response Assessment ................................ ................................ ................... 13 4.5.2 Recommendation for Combining Exposure Routes ................................ .............. 14 4.5.3 Cancer Classification ................................ ................................ ............................ 14 4.5.4 Summary

of Points of Departure Used in Risk Assessment ................................ .. 15 5.0 DI ETARY AND D RI NKING WATER RI SK ASSESSMENT ................................ ........ 16 5.1 Metabol ite/Degradate Residue Profile ................................ ................................ ............... 16 5.1.1 Summary

of Plant and Animal Metabolism Studies ................................ .............. 16 5.1.2 Comparison of Metabolic Pathway s ................................ ................................ ...... 16 5.1.3 Environmental Fate and Transport ................................ ................................ ......... 1 6 5.1. 4 Residues of Concern Summary and Rationale ................................ ....................... 1 7 5.2 Food Residue Profile ................................ ................................ ................................ .......... 18 5.2.1 Residues in Crops, L ivestock and Poultry ................................ ............................. 18 5.3 Water Residue Profile ................................ ................................ ................................ ........ 18Page 3 of 47

5.3.1 Estimated Drinking Water Concentrations ................................ ............................ 18 5.4 Dietary

and Drinking Water Exposure and Risk ................................ ............................... 19 5.4.1 Acute Dietary

Exposure and Risk ................................ ................................ .......... 19 5.4.2 Chronic Dietary

Exposure and Risk ................................ ................................ ...... 20 6.0 RESIDENT IA L EXPOSURE AND RI SK ................................ ................................ ........ 20 6 .1 Residential By stander Postapplication Inhalation Exposure ................................ ............. 20 6.2 Spray

Drift ................................ ................................ ................................ ......................... 21 7.0 AGGREGATE EXPOSURE AND RI SK AS SESSMENT ................................ ............... 21 7.1 Short - Term Aggregate Ri sk to Residential Applicators ................................ .................... 22 8.0 CUMULAT IVE RISK ................................ ................................ ................................ ....... 22 9.0 O CCUPATI ONAL EXPOSURE AND RI SK ................................ ................................ ... 23 9 .1 Exposure Scenarios ................................ ................................ ................................ ............ 23 9.2 Handler Exposure ................................ ................................ ................................ ............... 23 9.2 .1 Handler Exposure Scenarios ................................ ................................ .................. 23 9.2 .2 Handler Exposure Data ................................ ................................ .......................... 23 9.2 .3 Handler Exposure Assumptions ................................ ................................ ............. 23 9.2 . 4 Handler Exposure and Risk Estimates ................................ ................................ ... 24 9.3 Post Application Exposure ................................ ................................ ................................ . 25 9.3 .1 Post Application Exposure Scenarios ................................ ................................ .... 25 9.3 .2 Post Application Exposure Assumptions ................................ ............................... 25 9.3 .3 Post Application Exposure and Risk E stimates ................................ ..................... 26 9.3.4 Restricted Entry

Interval ................................ ................................ ........................ 26 10. 0 REFERENCES ................................ ................................ ................................ .................. 26 Appendix A. Toxicity

Pr ofile and Executive Summaries ................................ .............................. 28 A.1 Toxicity

Data Requirements ................................ ................................ .................. 2 8 A.2 Toxicity

Profiles ................................ ................................ ................................ ..... 2 9 A.3 Hazard Identification and Endpoint Selection ................................ ....................... 29 A.4 Execut ive Summaries ................................ ................................ ............................. 3 9 Appendix B. Metabolism ................................ ................................ ................................ .............. 45 Appendix C. Phy sical/Chemical Properties ................................ ................................ ................... 4 7 Appendix D. Review of Human Research ................................ ................................ ..................... 4 7Page 4 of 47

**1.0 EXECUTIVE SUMMARY**  This assessment provides information to support an amend ed Section 3 registration for the use of acibenzolar - S - methyl on low growing berries, crop subgroup 13 - 07G . This document addresses the exposures and risks associated with exposures from currently registered uses and the proposed new use s of acibenzolar - S - methyl. Italso assesses potential enhanced sensitivity of infants and children from dietary and/or residential exposure as required under the Food Quality Protection Act (FQPA) of 1996. **Use Profile** Acibenzolar - S - methyl is a p lant activator . Itis a member of the benzothiadiazole class of compounds. Acibenzolar - S - methyl is registered to Sy ngenta Crop Protection, Inc for use on a number of leafy

and fruiting v egetables crops and tobacco for the control or suppression of downy mildew, black rot, white rust, bacterial leaf spot and speck, and blue mold. P ermanent tolerances ranging from 0.1 - 3.0 ppm are established under §180.561(a) . Acibenzolar - S - m eth yl is also used in commercial seed treatment operations for protection against certain earl y season seed and seed - borne nematodes in cotton and on sod farms, golf courses, collegiate athletic fields, and lawns around commercial and industrial buildings to control turf diseases. Acibenzolar - S - methyl acts to protect plant s against invading pathogen s by induc ing a natural defense reaction that enhances the plants ability to ward off disease. Agricultural a pplications may be made using ground or aerial equipment. Acibenzolar - S - methyl is also registered for residential application to turf grass using handheld spray equipment. Proposed New Use s The Interregional Research Project No. 4 (IR - 4), on behalf of the Agricultural Experiment Stations of CO, NM, UT, ID, and WA, is requesting to amend the product label for Actigard® (EPA Reg. No. 100 - 922), a water - disp ersible granule (WG) formulation containing 50% acibenzolar - S - methyl to add a new use on the low growing berry crop subgroup 13 - 07G. Actigard® is proposed for multiple foliar applications on the low growing berry subgroup using ground or aerial equipment at a maximum of 0.023 lb ai/A per application, a maximum seasonal rate of 0.188 lb ai/A, and a 0 - day pre - harvest interval (PHI). **Hazard Identification** T he toxicology

database for acibenzolar - S - methyl is adequate for evaluating and characterizing toxicity

and selecting endpoints for purposes of this risk assessment. In subchronic and chronic oral studies in rats, dogs and mice, signs of mild regenerative hemoly tic anemia were consistently observed in all three species. Additional toxic effects observed in these studies included decreases in body weight, body weight gain and/or fo od consumption. No other significant treatment - related effects of toxicological concern were observed in these subchronic and chronic oral studies. No neurotoxic effects were seen at the highest dose tested in a subchronic neurotoxicity study in rats. In a 28 day

dermal toxicity

study in rats no sy stemic or dermal effects were seen at the limit dose.Page 5 of 47

In a developmental toxicity and developmental neurotoxicity studies in rats, treatment related effects were observed in fetuses at levels that were not toxic to the parent indicating increased sensitivity

of rat fetuses compared to adults. Increased sensitivity was not observed in a developmental toxicity

study in rabbits , or in 1 - and 2 - generation reproduction studies in rats. In a 28 day dermal develo pmental toxicity study in rats, no maternal or developmental toxicity was observed at dose levels up to 500 mg/kg/day , the highest dose level tested. Acibenzolar - S - methyl was classified by HED’s Hazard Identification Assessment Review Committee ( HIARC) as “not likely” to be a human carcinogen based on negative carcinogenicity findings in male and female rats and mice and negative results in an acceptable battery of genotoxicity

studies. Acibenzolar - S - methyl showed no significant toxicity in a battery of acute toxicity tests (Toxicity Category III or IV in all tests) but showed considerable skin - sensitivity . The end - use product (ActigardTM 50Wg) displayed no significant acute toxicity and no dermal sensitizing potential. An immunotoxicity

study required as part of new 40 CFR Part 158 data requirements for registration of a pesticide has been submitted and is being re viewed. Based on a preliminary review , the study

is acceptable and indicates no evidence of immunotoxicity . **Dose Response Assess ment**  Toxicological endpoints were selected for dietary /drinking water, occupational and residential exposure scenarios. Acute and chronic reference doses (RfDs) were selected for assessment of food and drinking water exposures. An acute RfD for the gene ral population was not selected because no effect attributable to a single day oral exposure was observed in animal studies. A cute and chronic RfD’s for females 13 - 49 were selected from a developmental neurotoxicity

toxicity study in rats based on changes in brain morphometrics in the cerebellum . A chronic RfD for the general population was selected on the basis of hemoly tic effects seen in a c hronic feeding study

in dogs and other co - critical toxicity studies . Toxicological endpoints for inhalation, dermal, and incidental oral exposures were selected from the developmental neurotoxicity

study in rats. A dermal absorption factor was selected from a dermal absorption study in rats. An uncertainty

factor of 100X was applied to endpoints selected for all exposure routes (10x for interspecies extrapolation, 10x for intraspecies variation). **Exposure/Risk Assessment and Risk Characterization** Risk assessments were conducted for dietary (food and water) and occupational exposure pathway s based on the request for new uses of acibenzolar - S - methyl on low growing berries. A new residential assessment is not required because the proposed new use does not include residential applications or exposures. Refined a cute and screening level chronic dietary and drinking water risk assessments for acibenzolar - S - methyl showed that dietary and drinking water exposure estimates are below HED’s level of concern for the general population and all population subgroups. Occup ational exposure and risk estimates indicate that worker handlerPage 6 of 47

and post - application exposures are not of concern at the maximum allowable application rates for the proposed new uses. Aggregate risks are not of concern. **Use of Human Studies** This risk assessment relies in part on data from studies in which adult human subjects were intentionall y exposed to a pesticide or other chemical. These studies , listed in Appendix D , have been determined to require a review of their ethical conduct . S ome of thes e studies are also subject to review b y the Human Studies Review Board. All of the studies used have received the appropriate review. **2.0 HED RECOMMENDATIONS**  **2.1 Data Deficiencies/Conditions of Registration**  Submission of a reference standard for the metabolite CGA 210007, and a revised Section F are required. An immunotoxicity study required as part of new 40 CFR Part 158 data requirements for registration of a pesticide has been submitted and is being reviewed. Based on a preliminary

review, the study

is acceptable and indicates no evidence of immunotoxicity . **2.2 Tolerance Considerations**  **2.2.1 Enforcement Analytical Method**  HPL C/UV Method AG - 617A is available for tolerance enforcement. This method has undergone a successful tolerance method validation by the Anal yt ical Chemistry Branch of BEAD. The method was forwarded to FDA for inclusion in PAM Volume II. Briefl y, the method consists of an initial hydrol ys is with NaOH to convert acib enzolar - S - methyl to CGA 210007 followed b y methanol extraction. Residues are then diluted with HCl and purified b y a series of solid - phase extraction steps. Prior to HPLC/UV anal ys is, residues are partitioned into ethyl acetate, dried down, and re - dissol ved in phosphoric acid. This method has a limit of quantitation (L OQ) of 0.02 ppm. The method includes optional detection via HPL C/MS, giving a means of residue confirmation. **2.2.2 International Harmonization**  There are no Codex, Mexican, or Canadian maximum residue limits for acibenzolar - S methyl in/on any

commodity . Therefore, international harmonization is not an issue. **2.2.3 Recommended Tolerances**  HED has examined the residue chemistry database for acibenzolar - S - methyl. Pending submission of a r evised Section F (see requirements under Proposed Tolerances), there are no residue chemistry

issues that would preclude granting a Section 3 registration for the requested use of acibenzolar - S - methyl on crop subgroup 13 - 07G, or the establishment of a tole rance for residues of acibenzolar - S - methyl, as follows:Page 7 of 47

**Table 1**  **. Tolerance Summary for**  **Acibenzolar -**  **S -**  **me thyl.**  **Comm odity**  **Established Tolerance (ppm)**  **Proposed**

**Tolerance (ppm) Recommended** 

**Tolerance (ppm) Comments;**  **Correct**

**Commodity Definition Tolerances**  **to be listed under 40 CFR §180.**  **561 (a):**  Berry , low

growing subgroup 1 3 - 07 G -- 0.15 0.15 Note to Registration Division: Tolerances have been established under 40 CFR §180.561 for residues of acibenzolar - S - methyl. HED continues to recommend that the appropriate tolerance expression for acibenzolar - S - methyl read as follows . Tolerances are established for residues of acibenzolar - S - methyl, benzo( 1,2,3)thiadiazole 7 - carbothioic acid - S - methyl ester, including its metabolites and degradates, in or on the commodities in the table below. Compliance with the tolerance levels specified below is to be determined b y measuring onl y those acibenzolar - S - methyl residues convertible to benzo(1,2,3)thiadiazole - 7 - carbox yl ic acid (CGA - 210007), expressed as the stoichiometric equivalent of acibenzolar - S - methyl, in or on the commodity . **2.2.4 Revisions to Petitioned**  **- For Tolerances**  There are no revisions to the petitioned for tolerance. **3 .0**  **INGREDIENT PROFILE 3.1**  **Chemical Identity Table**  **2 .**  **Acibenzolar -**  **S -**  **Methyl Nom**  **enclature** . Chemical structure

Common name acibenzolar - S - methyl Com pany experimental name CGA - 245704 IUPAC name benzo [1,2,3]thiadiazole - 7 - carbothioic acid - S - me thyl ester CAS name S - methyl, 1,2,3 - bensothiadiazole - 7 - carbothioate CAS registry number 135158 - 54 - 2 End - use product (EP) Actigard® 50 WG (EPA Reg. No. 100 - 922) Structure of metabolite CGA - 210007 1,2,3 - benzothiadiazole - 7 - carboxylic acid N N S O S C H 3

N N S C O O HPage 8 of 47

**3.2 Physical/Chemical Characteristics**  A detailed description of the phy siciochemical properties of acibenzolar - S - methyl is provided in Appendix B . Acibenzolar - S - methyl exhibits relatively low solubility

in water and higher solubility

in solvents. It has a very low vapor pressure. It does not present significant concerns for bioaccumulation based on its lipophilicity and mammalian metabolism studies. **3.3 Pesticide**  **Use Pattern 3.3.1**  **Registered Products** There are currently

7 active acibenzolar - S - methyl registrations including four Section 3 registrations and five special local needs (SLNs) or 24(c) registration s. **Table 3 . Summary Report of Supported**  **Registered Acibenzolar**  **- S**  **- me**  **thyl Products Reg #**  **Nam e**  **Company Nam e**  **%AI** 100 - 921 Acibenzolar - S - Methyl Technical Syngenta Crop Protection, Inc. 99 100 - 922 Actigard 50 WG 50 100 - 1363 Bion 500 SF 54 100 - 1364 Daconil Action 0.11 FL030003 Actigard 50 WG Plant Activator 50 GA020005 50 NC020007 50 SC030004 50 VA030002 50 **3.3. 2**  **Proposed New Uses** The Interregional Research Project No. 4 (IR - 4), on behalf of the Agricultural Experiment Stations of CO, NM, UT, ID, and WA, is requesting to amend the product label for Actigard® (EPA Reg. No. 100 - 922), a water - dispersible granule (WG) formulation contai ning 50% acibenzolar - S - methyl to add a new use on the low growing berry crop subgroup 13 - 07G. Actigard® is proposed for multiple foliar applications on the low growing berry subgroup using ground or aerial equipment at a maximum of 0.023 lb ai/A per appli cation, a maximum seasonal rate of 0.188 lb ai/A, and a 0 - day pre - harvest interval (PHI). **Table 4**  **. Proposed New Use of Acibenzolar -**  **S -**  **Methyl on Low Growing Berries Applic.**  **Timing, Type, and**  **E quip.**  **Form ulation** 

**[EPA Reg #**  **] Applic.** 

**Rate (lb ai/A)**  **Max. No.**

**Applic. per Season**  **Max.**

**Seasonal App**  **Rate (lb ai/A)**  **PHI**

**(days) Use Directions and Limitations**  Postemergence Foliar or banded Ground /Aerial 50% WG

[100 - 922] 0.016 -

0.023 8 (Implied) 0.188 0 Actigard® should be applied to healthy, actively growing plants. Do not apply prior to thinning or within 5 days of transplanting. Do not apply Actigard® to plants that are stressed due to drought, excessive moisture, cold weather, or herbicide injury, etc. Avoi d spray overlap as injury may occur .Page 9 of 47

**3.4 Anticipated Exposure Pathways**  Dietary (food and water) and residential exposures are expected based on existing uses of acibenzolar - s - methyl on low growing berries . A residential exposure assessment is not required for this assessment because there are no residential uses or exposures associated with the proposed new use. The short - term aggregate exposure assessment, which takes into account residential exposure p lus average exposure levels to food and water, has been updated to incorporate dietary

exposure from the proposed new uses on low growing berries . **3.5 Considerations of Environmental Justice**  Potential areas of environmental justice concerns, to the extent possible, were considered in this human health risk assessment, in accordance with U.S. Executive Order 12898, "Federal Actions to Address Environmental Justice in Minority Populations and Lo w - Income Populations," http://www.eh.doe.gov/oepa/guidance/justice/eo12898.pdf ). As a part of every

pesticide risk assessment, OPP considers a large variety of consumer subgroups accord ing to well - established procedures. In line with OPP policy , HED estimates risks to population subgroups from pesticide exposures that are based on patterns of that subgroup’s food and water consumption, and activities in and around the home that involve pesticide use in a residential settin g. Whenever appropriate, non - dietary exposures based on home use of pesticide products and associated risks for adult applicators and for toddlers, youths, and adults entering or playing on treated areas postapplication are evaluated. Further considerati ons are currently in development, as OPP has committed resources and expertise to the development of specialized software and models that consider exposure to by standers and farm workers as well as lifestyle and traditional dietary patterns among specific subgroups. **4 .0**  **HAZARD CHARACTERIZATION/ASSESSMENT 4.1**  **Toxicology Studies Available for Analysis** T he toxicology

database for acibenzolar - S - methyl is adequate for evaluating and characterizing acibenzolar - S - methyl toxicity and selecting endpoints for purposes of this risk assessment. An immunotoxicity study required as part of new 40 CFR Part 158 data requirements for registration of a pesticide has been submitted and is being reviewed. Based on a preliminary

review, the study

is acceptable and indicates no evidence of immunotoxicity . **4.2 Absorption, Distribution, Metabolism and Excretion**  The absorption, distribution, metabolism, and excretion of acibenzolar - S - methyl were studied in rats. In the rat metabolism study [U 14 C Phenyl ] acibenzolar - S - methyl was administered b y stomach tube to male and female rats at single doses of 0.5 (low dose) or 100 mg/kg (high dose) or at a single dose of 0.5 mg/kg following 14 repeated daily doses of 0.5 mg/kg of nonlabeled acibenzolar - S - me thyl. For all treatment groups, acibenzolar - S - methyl was rapidly and nearly

completely absorbed (>90% of the administered dose) from the gastrointestinal tract into the generalPage 10 of 47

circulation. Peak blood levels were reached at 15 - 30 minutes after the lo w dose and at 4 - 8 hours after the high dose. Tissue levels of radioactivity were highest in blood, liver, kidney , skeletal muscle, heart and lungs. Radioactivity was rapidly

excreted predominantl y in the urine (88 - 95% within 48 hours). Only

a small amount of radioactivity was excreted in the feces (3.1 - 4.5% within 48 hours). Insignificant radioactivity was excreted in the expired air. There was no significant enterohepatic circulation. After 7 day s, levels of radioactivity remaining in the tissues wer e extremel y low. Absorption, distribution, pharmacokinetic and excretion profiles were largel y independent of dose level and sex . Three urinary

fractions were identified. Fraction U3 (79 - 92 % of the administered dose) was identified the carboxylic acid derivative of the parent . Fraction U2 (0.4 - 2% ) was identified as the corresponding glycine derivative of the carbox yl ic acid (fraction U3) derivative. Fraction U1 (1 - 4.5% ) was a mixture of at least three polar compounds that were not identified due to insufficient sample material. No parent compound was detected in the urine. Radioactivity in the feces represented <5% of the administered dose. No single metabolite in the feces exceeded 3% of the administered dose. Fraction F2 (0.7 - 3 % of the administered dose) was identified as the carbox yl ic acid derivative of the parent . Fraction F3 (0.1 - 1%) was identified as parent compound. Frac tion F1 (0.2 - 0.4%) was a very polar compound(s) that was not further identified. The metabolic fate of acibe nzolar - S - methyl was independent of dose level, sex, and pretreatment. In a dermal absorption study in rats , acibenzolar - S - methyl was applied dermally to a shaved dorsal area of 10 cm2 at two dose levels of 10 μg/cm2 and 100μg/cm2 to two groups of animals consisting of six animals/group. T wo animals from each group were sacrificed at 4, 8, and 24 hours after dosing and the amount of radioactivity in blood, urine , feces , liver, kidneys, spleen, and remaining carcass was determined. Amount of radioac tivity remaining in/ on the skin and on the cover wa s also determined . For the groups for which the skin was washed after 8 hours, mean dermal absorption values were 43.55% and 5.92%, and mean radioactivity remaining at the skin were 46.64%, and 90.22%, respectivel y at 10 μ g/cm2 and 100 μ g/cm2 dose levels. Given the uncertainty regarding actual dermal deposition under actual field conditions, it is considered appropriate to derive an estimate of dermal absorption based on the results from the low dose gro up (10 μ g/cm 2 ), since percent dermal absorption was greatest at this dose level. Based on the likely

worker exposure time frame, it is considered most appropriate to adopt the dermal absorption value calculated for the group of animals for which the skin site was washed after 8 hours. Mean dermal absorption for this group of animals was 43.55%. (TXR 0054866 and/or MRID 47430503 ) **4.3 Toxicological Effects**  In subchronic and chronic oral studies in rats, dogs and mice, signs of mild regenerative hemoly tic anemia were consistently observed in all three species. These signs frequentl y included decreased ery

throcy te counts, decreased hemoglobin, decreased hematocrit, increa sed reticulocy te counts, increased hemosiderosis in the spleen, liver and/or bone marrow, extramedullary

hematopoiesis in the spleen, and increased spleen weights in both males and females. A compensatory response (increased erythrocy te production) regularly

followed the initial anemia. Additional toxic effects observed in these same studies included decreases in body weight, body weight gain and/or food consumption. No other significant treatment - relatedPage 11 of 47

effects of toxicological concern were observed in these subchronic and chronic oral studies. In a 28 - day dermal study

in rats, no sy stemic or dermal effects were observed at dose levels up to 1000 mg/kg/day , the limit dose. No neurotoxic effects were observed at any dose in a subchronic neurotoxicity

s tudy in rats . T reatment - related developmental malformations, anomalies and variations were observed in a developmental toxicity study in rats at or below the no observable adverse effect level (NOAEL ) for maternal toxicity . At the highest dose tested i n this study (400 mg/kg/day), both maternal toxicity

(hemorrhagic perineal discharge) and considerable developmental toxicity (including total litter resorptions, fetal malformations, anomalies and variations) were observed. The fetal malformations noted at this dose included treatment - related effects on nervous sy stem tissues (hydrocephaly , craniorachisis and anophthalmia/ microphthalmia). At the next lower dose tested (200 mg/kg/day), treatment - related visceral malformations and skeletal variations were demonstrated in the absence of significant maternal toxicity . A similar increased sensitivity of fetuses or pups (as compared to adults) was not observed in a developmental toxicity study in rabbits or in 2 - generation and 1 - generation (range - finding) stu d ies in rats. In a dermal developmental toxicity

study in rats, no maternal or developmental toxicity was observed at dose levels up to 500 mg/kg/day , the highest dose tested. In a battery

of mutagenicity

studies, results were negative in all studies e xcept in an in vitro chromosome aberration study in Chinese hamster ovary (CHO) cells, in which there was evidence of a clastogenic response in the absence of S - 9 activation. In a 2 - ye ar chronic toxicity /carcinogenicit y study in rats and an 18 - month c arcinogenicit y study in mice, acibenzolar - S - methyl was negative for carcinogenicit y when administered at dose levels adequate for the testing of carcinogenic potential. Acibenzolar - S - methyl showed no significant toxicity in a battery

of acute toxicity tests (Toxicity Category III or IV in all tests). Considerable skin sensitizing (contact allergenic) potential was demonstrated in a dermal sensitization study in guinea pigs. T he formulated end use product (ActigardTM 50 WG) also demonstrated no signific ant acute toxicity ; additionally , the end - use product did not show dermal sensitization in guinea pigs. The complete toxicity

profile for acibenzolar - S - methyl is provided in Appendix A . **4.4 Safety Factor for Infants and Children (FQPA Safety Factor)**  The FQPA factor for increased susceptibility to infants and children is reduced to 1x based on the following cons iderations. The toxicology data base for acibenzolar - S - methyl is complete and adequate for assessing inc reased susceptibility under FQPA. The pre - and postnatal toxicity database for acibenzolar - S - methyl includes developmental toxicity studies in rats and rabbits, a DNT study in rats, and a two - generation reproduction toxicity study in rats. Based on the de velopmental toxicity in rats and the developmental neurotoxicity studies in rats , the re is concern for increased qualitative and/or quantitative susceptibility following in utero exposure to acibenzolar - S - methyl. However, the degree of concern for the inc reased susceptibility seen in these studies is low as there are no residual uncertainties with regard to pre - and/or postnatal toxicity since NOAELs and L OAELs have been identified for all effects of concern, a clear dosePage 12 of 47

response has been well defined, and the points of departure ( PODs ) selected for risk assessment are protective of the fetal/offspring effects. Additionally , the dietary and residential risk assessment s are conservative and will not underestimate dietary e xposure and there are no residual uncertainties in the exposure database. Further discussion may be found in the following sections. **4.4.1 Completeness of the Toxicology Database**  The toxicity

database for acibenzolar - S - methyl is sufficient for a full hazard evaluation and is considered adequate to evaluate risks to infants and children. Acceptable studies are available for developmental , reproduction, developmental and subchronic neurotoxicity . The i mmunotoxicity study required under 40 CFR Part 158 data requirements has been submitted and is being reviewed . Based on a preliminary review, the study

is acceptable and indicates no evidence of immunotoxicity . **4.4.2 Evidence of**  **Neurotoxicity** In a developmental neurotoxicity study a maternal LOAEL was not observed. The maternal NOAEL is 326 mg/kg/day , the highest dose tested . No treatment - related offspring effects were seen on survival, clinical signs, FOB, developmental land marks, brain weights or neuropathology . No conclusions can be drawn on the eff ects of the compound on motor activity due the lack of consistency in the observations. Significant m orphometric changes (decreased thickness of the molecular layer of the cerebellum ) w ere observed in male offspring on PND 63 at 82 mg/kg/day . At the high dose, treatment - related offspring effects included decreased body weights , increased auditory startle response and increased thickness in the corpus callosum in females . In subchronic neurotoxicity study in rats no compound - related effects wer e observed in the FOB, motor activit y, gross or neuropathology at the highest doses ( 575/628 mg/kg/day , M/F ) tested . **4.4.3 Evidence of Sensitivity/Susceptibility in the Developing or Young Animal**  In the rat developmental toxicity study treatment related visce ral malformations and skeletal variations were observed in fetuses at 200 mg/kg/day , the NOAEL for maternal toxicity . In the developmental neurotoxicity study , offspring toxicity

was observed at 82 mg/kg/day while no maternal toxicity

was observed at 326 mg/kg/day , the highest dose tested. Additional developmental toxicity

studies in rats and rabbits and reproduction studies in rats provided no indication of increased susceptibility of rat or rab bit fetuses or neonates compared to adult animals. Based on the results of developmental toxicity and developmental neurotoxicity studies in rats the re is concern for increased qualitative and/or quantitative susceptibility following in utero exposure to acibenzolar - S - methyl. However, the degree of concern for the increased susceptibility

seen in these studies is low as there are no residual uncertainties with regard to pre and/or postnatal toxicity since NOAELs and LOAEL s have been identified for all ef fects of concern, a clear dose response has been well defined, and the points of departure ( PODs ) selected for risk assessment are protective of the fetal/offspring effects. There are no residual uncertainties with regard to pre - and/or postnatal toxicity .Page 13 of 47

**4.4.4 Residual Uncertainty in the Exposure Database**  There are no residual uncertainties in the exposure database. The dietary risk assessment is conservative and will not underestimate dietary and/or no n - dietary residential exposure to acibenzolar - S - methyl . **4.5 Toxicity Endpoint and Point of Departure**  **4.5.1 Dose**  **- Response Assessment**  Toxicity endpoints and points of departure (PODs) for dietary (food and water), occupational, and residential exposure scenarios are summarized below. A detailed description of the studies used as a basis for the selected endpoints are presented in Appendix A. An acute POD of 8.2 mg/kg/day (NOAEL) was selected for all populations from an developmental neurotoxicity stu dy in rats based on changes in brain morphometrics in the cerebellum at the LOAEL of 82 mg/kg/day . An uncertaint y factor (UF) of 100x (10x to account for interspecies extrapolation and 10x for intraspecies variation) was applied to the NOAEL to obtain an acute reference dose (aRfD) of 0.1 mg/kg/day . Since the FQPA factor has been reduced to 1X, the acute population adjusted dose (aPAD) is equivalent to the aRfD. The developmental neurotoxic effects of concern are presumed to occur after a single exposure and are relevant to females aged 13 - 50 years old since they occur in utero and to young children based on post - natal effects. [Note: In earlier risk assessment s (D361893, 3/3/09) , only the subpopulation females 13 - 50 years old was assessed for acute dietary exposure based on in utero effects. However, based on a reevaluation of the DNT study regarding the potential for post - natal toxicity , the acute dietary anal ys is has been expan ded to all populations in order to include children for this and more recent previous assessment s (D379425, 8/19/10) .] A chronic POD of 8.2 mg/kg/day (NOAEL) was selected for females 13 - 49 and young children years old from a developmental neurotoxicity study in rats based on changes in brain morphometrics in the cerebellum at the LOAEL of 82 mg/kg/day . A UF 100x (10x to account for interspecies extrapolation and 10x for intraspecies variation) was applied to the dose to obtain a chronic reference dose (cR fD/cPAD) of 0.1 mg/kg/day . The effects observed in this study can occur after single or multiple doses. Selection of a chronic endpoint from this study ensures that the chronic exposure assessment is protective of f emales of reproductive age and their of fspring. [Note: In earlier risk assessment s (D361893, 3/3/09) , only the subpopulation females 13 - 50 years old was assessed for chronic dietary exposure based on in utero effects observed in the DNT study . However, based on a reevaluation of the DNT study regarding the potential for post - natal toxicity , the chronic dietary anal ys is the chronic dietary exposure analy sis has been revised to include an assessment of dietary risks young children using the NOAEL from the DNT study for this and more recent previous assessments (D379425, 8/19/10). ] A chronic POD of 25 mg/kg/day (NOAEL) of 25 mg/kg/day was selected for adult males and females (50+ ye ars) based on hemol yt ic effects seen in chronic dog study and other co -Page 14 of 47

cri tical toxicity

studies. A UF 100x (10x to account for interspecies extrapolation and 10x for intraspecies variation) was applied to the dose to obtain a chronic reference dose (cRfD/cPAD) of 0.25 mg/kg/day . The NOAEL of 25 mg/kg/day , taken from the chron ic dog study , is based on the totality

of the database with a LOAEL of 105 mg/kg/day (the lowest LOAEL is from the reproduction study) based on hemolytic anemia with compensatory response. The NOAEL and LOAEL recommended for use in the risk assessment com e from different studies conducted in different species . Ty

pically , a combination of NOAEL s/LOAELs from studies conducted in different species is not recommended. However, in the case of acibenzolar - S - methyl the same endpoint (hemoly

tic anemia) is observ ed across species at a relativel y close range of LOAE Ls (105 - 312 mg/kg/day). Therefore, the HED Tox icity Science Advisory

Committee concluded that, in this instance, it would be appropriate to base the point of departure for the risk assessment on the NOA EL from the dog chronic toxicity study using the LOAEL from the reproduction study in rats for the basis of the endpoint (hemoly tic anemia). Short - term incidental oral and short - and intermediate term dermal and inhalation PODs of 8.2 mg/kg/day

were selected from the developmental neurotoxicity in rats. The selected endpoint is protective of offspring effects from dermal exposure. Two acceptable dermal toxicity studies, including a dermal developmental study are available. However s in ce neurological effects observed in the oral developmental neurotoxicity study are not measured in the dermal stud ies , the findings from the se stud ies may not be protective of neurological effects. A dermal absorption factor (DAF) is applied when dermal e xposure endpoints are selected from oral toxicity studies. The dermal factor converts the oral dose to an equivalent dermal dose for the risk assessment. A dermal absorption factor of 45 % was used for the dermal exposure assessment based on a dermal absorp tion study in rats An MOE 100 is required for incidental oral, dermal and inhalation exposure scenarios based on the conventional uncertainty factor of 100 ( 10x for interspecies extrapolation and 10x for intraspecies variation). **4.5.2 Recommendations for**  **Combining Exposure Routes** When there are potential residential exposures to the pesticide, aggregate risk assessment must consider exposures from three major sources: oral, dermal and inhalation exposures. T here is potential residential post - application exposure to adults via the dermal route and to children via dermal and i ncidental oral routes of exposure. Dietary , dermal and incidental oral endpoints for acibenzolar - S - methyl are based on common toxicological effects for females 13 - 49 years old and children. Therefore, exposure s from food and non - food oral and dermal routes should be aggregated for those subpopulations for this assessment. In addition, d ermal , and inhalation exposures should be aggregated for acibenzolar - S - methyl, bec ause the same oral toxicity endpoint was selected for both exposure routes . **4.5.3 Classification of Carcinogenic Potential**  A cibenzolar - S - methyl has been designated b y HED as “Not Likel y to be C arcinogenic to Humans” in accordance with the 2005 EPA Guidelines for Carcinogenic Risk Assessment based on negative carcinogenicity studies in rats and mice and on generally negative mutagenicity studies.Page 15 of 47

**4.5.4 Summary of**  **Points of Departure Used in Risk Assessment**  Toxicological doses/endpoints selected for the acibenzolar - S - methyl risk assessment are provided in Tables 5 and 6 . **Table 5**  **. Summary of Toxicological Doses and Endpoints for Acibenzolar**  **- S**  **- me**  **thyl for Use in Dietary Hum**  **an Health Risk Assessments**  **Exposure/ Scenario**  **Point of**

**Departure Uncertainty/F** 

**QPA Safety Factors**  **RfD, PAD,**

**LOC for Risk Assessment**  **Study and Toxicological Effects**

Ac ute Dietary (General Population) NOAEL =

8.2 mg/kg/day UF A= 10x

UF H =10x FQPA SF= 1x Ac ute RfD =

0.082 mg/kg/day aPAD = 0.082 mg/kg/day Developmental Neurotoxicity Toxicity - Rat

Developmental LOAEL = 82 mg/kg/day based on changes in brain morphometrics in the cerebellum in offspring. Maternal LOAEL = was not observed NOAEL = 326.2 mg/kg/day HDT Chronic Dietary (Females 13 - 49 years & Young Children) NOAEL =

8.2 mg/kg/day UF A= 10x .

UF H =10x FQPA SF= 1x Ac ute RfD =

0.082 mg/kg/day aPAD = 0.082 mg/kg/day Developmental Neurotoxicity Toxicity - Rat

Developmental LOAEL = 82 mg/kg/day based on changes in brain morphometrics in the cerebellum in offspring. Maternal LOAEL = was not observed NOAEL = 326.2 mg/kg/day HDT Chronic Dietary ( Adult Males and Females 50+ yrs ) NOAEL =

25 mg/kg/day UF A= 10x

UF H =10x FQPA SF= 1x Chronic RfD =

0.25 mg/kg/day cPAD = 0.25 mg/kg/day Chronic Toxicity - Dog; Co - critical;

Chronic/Cancer - Rat & Mouse, Reproduction Toxicity - Rat LOAEL = 105 mg/kg/day based on hemolytic anemia with compensatory response. Incidental Oral NOAEL=

8.2 mg/kg/day UF A= 10x

UF H =10x Occupational

LOC for MOE = 100 Developmental Neurotoxicity Toxicity - Rat

Developmental LOAEL = 82 mg/kg/day based on changes in brain morphometrics in the cerebellum in offspring. Maternal LOAEL = was not observed NOAEL = 326.2 mg/kg/day HDT Cancer (all routes) A “not likely” human carcinogen Point of Departure (POD) = A data point or an estimated point that is derived from observed dose - response data and used to mark the beginning of extrapolation to determine risk associated with lower environmentally relevant human exposures. NOAEL = no observed adverse effect level. LOAEL = lowest observed adverse effect level. UF = uncertainty factor. UF A = extrapolation from animal to human (i nterspecies). UF H = potential variation in sensitivity among members of the human population (intraspecies). UF L = use of a LOAEL to extrapolate a NOAEL. UF S = use of a short - term study for long - term risk assessment. UF DB = to account for the absence o f key date (i.e., lack of a critical study). FQPA SF = FQPA Safety Factor. PAD = population adjusted dose (a = acute, c = chronic). RfD = reference dose. MOE = margin of exposure. LOC = level of concern. N/A = not applic able.Page 16 of 47

**Table 6**  **. Summary of Toxicological Doses and Endpoints for Acibenzolar**  **- S**  **- me**  **thyl for Use in Occupational**  **Hum an Health Risk Assessments**  **Exposure/ Scenario**  **Point of**

**Departure Uncertainty** 

**Factors LOC**  **for Risk**

**Assessment Study and Toxicological Effects**  Dermal Short (1 - 30 days) and Intermediate (1 - 6 months) Term DAF = 4 0% NOAEL=

8.2 mg/kg/day UF A= 10x

UF H =10x Occupational

LOC for MOE = 100 Developmental Neurotoxicity Toxicity - Rat

Developmental LOAEL = 82 mg/kg/day based on changes in brain morphometrics in the cerebellum in offspring. Maternal LOAEL = was not observed NOAEL = 326.2 mg/kg/day HDT Inhalation Short (1 - 30 days) and Intermediate (1 - 6 months) Term NOAEL=

8.2 mg/kg/day UF A= 10x

UF H =10x Occupational

LOC for MOE = 1 00 Developmental Neurotoxicity Toxicity - Rat

Developmental LOAEL = 82 mg/kg/day based on changes in brain morphometrics in the cerebellum in offspring. Maternal LOAEL = was not observed NOAEL = 326.2 mg/kg/day HDT Cancer ( all routes ) A “not likely” human carcinogen DAF = Dermal Absorption Factor . Since no inhalation absorption data are available, toxicity by the inhalation route is considered to be equivalent to the estimated toxicity by the oral route of exposure (100% absorption factor). **5.0 DIETARY AND DRINKING WATER EXPOSURE**  **AND RISK ASSESSMENT**  **5.1 Metabolite/Degradate Residue Profile**  **5.1.1 Summary of Plant Metabolism Studies**  No new plant metabolism studies were submitted. Adequate metabolism studies with [phenyl - U 14 C]acibenzolar - S - methyl on lettuce, tomato, rice, wheat, and tobacco have previously been submitted and reviewed. The metabolic pathway for acibenzolar - S - methyl was similar in all tested crops and involves the h ydrol ys is of the S - methyl ester to form the carbox yl ic aci d metabolite (CGA - 210007). Further hydrox yl ation of CGA - 210007 at the C - 4 and C - 5 positions results in the formation of the minor metabolites CGA - 324041 (all crops tested) and CGA 323060 (lettuce and tomato), which may also form acyl sugar conjugates. **5.1.2 Comparison of Metabolic Pathways**  CGA 210007 represented the vast majority of the urinary

radioactivity

in the rat metabolism study

and is therefore part of the total toxic mammalian exposure . CGA 210007 was also the principal metabolite in plant studies. As noted below , residues of the 4 - and 5 - hydroxy metabolites (CGA - 324041 and CGA - 323060) of CGA - 210007 should be considered to be equivalent to that of the parent compound acibenzolar - S - methyl. **5.1.3 Environmental Fate and Transport**  Laboratory

and field studies indicate that acibenzolar - S - methyl does not have a high potential to leach to ground water nor move to surface water based on its short resident times inPage 17 of 47

soil and soil water. The major degradate observed was benzo[1,2,3]thiadi azole - 7 - carbox yl ic acid (CGA 210007) in the h ydrolysis and soil photolysis studies (up to 100 percent). CGA - 210007 has a longer soil residence time, lower Freundlich Kd adsorption values, and is much more water soluble than acibenzolar - S - methyl, thereby e xhibiting greater potential than the parent for lateral or vertical movement after application. Acibenzolar - S - methyl mainly degrades by

photoly sis and microbial action. Aqueous and soil photoly sis irradiated half lives for parent are 0.6 hours and 1 day , respectivel y. The aerobic soil metabolism, anaerobic aquatic metabolism and aerobic aquatic half lives are 5.3 hours, 4.0 and < 1 day (s) respectivel y. Field dissipation half - lives ranged from < 1 to 27 days. Under aerobic soil, aerobic aquatic, and anae robic aquatic conditions acibenzolar quickly degrades to CGA 210007. CGA 210007 is in its anionic form at environmentally

significant pH's (pH 6 - 9) and appears to be more soluble than its parent, although measured at much lower pH’s (approx. 225 ppm at 25 0C pH 3.6 - 3.8 v. 7.7 ppm at 250C pH 7.5 - 7.9). CGA 210007 has longer resident field times than parent (aerobic soil half - life at 16.5 day s) and based on the batch equilibrium and aged soil column leaching studies, is considerabl y more mobile. Acibenzolar is not considered a bioconcentrate nor is it highly bioaccumulative. **5.1.4 Residues of Concern**  **Summary and Rationale** The HED Metabolism Assessment Review Committee ( MARC ) determined that the residues of concern in plants for tolerance expression are residue s convertible to benzo(1,2,3)thiadiazole - 7 - carbox yl ic acid (CGA - 210007), expressed as acibenzolar - S - methyl. For purposes of risk assessment, residues of the 4 - and 5 - hydrox y metabolites (CGA - 324041 and CGA - 323060) of CGA - 210007 should also be considered. For this purpose, the toxicity of CGA - 324041 and CGA - 323060 should be considered to be equivalent to that of the parent compound acibenzolar - S - methyl. These residues occur in significant quantities in lettuce and tomato. At this time, acibenzolar - S - methyldoes not have common metabolites with other agrochemicals. The residues of concern for drinking water are the same as those listed in the tolerance expression; that is, residues convertible to CGA - 210007. Table 7 provides a summary of the MARC decisions regarding residues of concern for acibenzolar - S - methyl . **Table 7**  **. Summ**  **ary of MARC D**  **ecisions for Acibenzolar**  **- S**  **- methyl**  **Matrix Residues of Concern**  **1 For Risk Assessment**  **2 For Tolerance Expression**  Plants Residues of acibenzolar - S - methyl, benzo(1,2,3)

thiadiazole - 7 - carbothioic acid - S - methyl ester, convertible to benzo(1,2,3)thiadiazole - 7 - carboxylic acid (CGA - 210007), expressed as acibenzolar - S - methyl & 4 - hydroxy CGA - 210007 (CGA - 323060) 5 - hydroxy CGA - 210007 (CGA - 324041) Residues of acibenzolar - S - methyl, b enzo(1,2,3) thiadiazole - 7 - carbothioic acid - S - me thyl ester, convertible to benzo(1,2,3)thiadiazole - 7 - carboxylic acid (CGA - 210007), expressed as acibenzolar - S - methyl 1 The residues of concern for drinking water are the same as those listed in the tolerance expression 2 If a future petition is received for rice, residues of CGA - 379019 should be included in the risk assessmentPage 18 of 47

**5.2 Food Residue Profile**  **5.2.1 Residues in Crops**  The registrant submitted crop field trial data from field trial conducted on strawberry as the representative crop of the low growing berry subgroup 13 - 07G. Crop field trials conducted on strawberry

as the representative are in accordance with the guideli ne for strawberry as the representative crop of the low growing berry subgroup 13 - 07G. The number and locations of field trials are in accordance with OPPTS guideline requirements . The trials reflect the proposed maximum seasonal rate and pre - harvest int erval (PHI), and there are adequate storage stability data to validate sample storage conditions and duration. The majority of the tank mixes contained an adjuvant. T he maximum residues of acibenzolar - S - methyl (i.e., residues convertible to CGA - 210007) w ere 0.087 ppm in/on strawberry . The residue data w ere entered into the OECD MRLCalculator. The available strawberry field trial data are deemed adequate to support a tolerance of 0.15 ppm for residues of acibenzolar - S - methyl in/on the low growing berry subgroup 13 - 07G. Adequate confined rotational crop data have been submitted previously . Based on the available field rotational crop data for wheat, turnip, and lettuce, tolerances are not needed for rotational crops provided a 30 - day plant - back interval is established fo r all crops not listed as target crops. The existing rotational crop restrictions on the Actigard® label are adequate. **5.3 Water Residue Profile**  **5.3. 1**  **Estimated Drinking Water Concentrations** Drinking water residues provided b y the Environmental Fate and Effects Division (EFED) (D35 3745 , R. Miller, 9 /2 /10) were incorporated directly into the acute and chronic dietary anal ys es . T he surface water estimates were based on EFED’s Tier II model Pesticide Root Zone Model . The residues of concern for drinking water include acibenzolar - S - methyl and residues convertible to CGA - 210007. The estimated drinking water concentrations (EDWCs) for use of acibenzolar - S - methyl on cucumbers were used for the drinking water assessment because this us e represents the highest use rate of the existing and proposed registered uses. The SCI GROW estimated ground water drinking water concentrations for acibenzolar is not expected to exceed 1.25 x 10 - 5 μg/Land CGA 210007 is not expected to exceed 8.12 x 10 - 2 μg/L . Surface water EDWCs for acibenzolar - S - methyl, which were used in the exposure assessment, are provided in Table 8 . CGA - 210007 drinking water residues were included in the dietary exposure assessment as acibenzolar - S - methyl equivalents.

**Table 8**  **. EDWCs for use in screening the hum an health risk assessment (**  **FL Cucumber )**  **Drinking Water Source**  **Chemical 1**  **- in**  **- 10 year**  **annual acute conc.**  **(μg/L) 1**  **- in**  **- 10 year avg** 

**annual (chronic) concentration**  **(μg/L) 30**  **- year avg. annual** 

**concentration (μg/L)**  Surface Water Acibenzolar - S - Methyl 1 0.7 2 0. 02 0.0 2 Surface Water CGA 210007 2 20.02 8.09 4.68 1 Aerial application method 2 Degradation product formed after applicationPage 19 of 47

**5 .**  **4 Dietary and Drinking Water Exposure and Risk**  Refined acute and screening - level chronic dietary and drinking water exposure and risk assessments were conducted using the Dietary Exposure Evaluation Model with the Food Commodity

Intake Database (DEEM - FCIDTM). Dietary risk assessment incorporates both exposure and toxicity

of a given pe sticide. For acute and chronic dietary assessments, the risk is expressed as a percentage of a maximum acceptable dose (i.e., the dose which HED has concluded will result in no unreasonable adverse health effects). This dose is referred to as the populat ion adjusted dose (PAD). The PAD is equivalent to the reference dose (RfD) divided b y the additional Safety Factor, if applied. For acute and non - cancer chronic exposures, HED is concerned when estimated dietary risk exceeds 100% of the PAD. Available lettuce metabolism data for a 7 - day PHI were used to estimate the relative abundance of the h ydroxy metabolites (CGA - 323060 and CGA - 324041) to the residues of acibenzolar - S - methyl convertible to the benzo(1,2,3) thiadiazole - 7 - carboxylic acid (CGA 210007). Based on the lettuce metabolism data, a factor of 1.5 X was applied to estimate s of acibenzolar - S - methyl residues to account for all of the residues of concern for dietary risk (including CGA - 323060 and CGA - 324041 ) . **5 .**  **4 .1**  **Acute Dietary and Drinking Water Analysis**  A refined (probabilistic) acute dietary exposure analy sis was performed for the general population and all population subgroup s . The acute anal ys is assumed a distribution of residues based on field trial data. Empirical and DEEM default p rocessing factors were used to modify the field trial data. M aximum screening - level percent crop treated estimates were used for commodities for which data were available. If no percent crop treated data were available, 100% crop treated was assumed. Th e acute anal ys is incorporated the entire time distribution of EDWCs adjusted to account for CGA - 210007. Acute dietary risk estimates are not of concern for general population or other population subgroups. The subgroup with the highest risk estimate was children 3 - 5 years old with a 99.9 th percentile acute exposure estimates of 3 5 % of the aPAD. The 99.9 th percentile aPAD for the general U.S. population was 1 7 %. **Table 9 : Summary of Acute Dietary Exposure and Risk for Acibenzolar**  **- S**  **- Methyl**  **Population Subgroup**  **aPAD (m g/kg/day)**  **Acute (99 th**  **Percentile) Exposure (mg/kg/day)**  **% aPAD** General U.S. Population

0.082 0.014091 17 All Infants (< 1 year old) 0.012951 16 Children 1 - 2 years old 0.025042 31 Children 3 - 5 years old 0.028536 35 Children 6 - 12 years old 0.019902 24 Youth 13 - 19 years old 0.015095 18 Adults 20 - 49 years old 0.009060 11 Adults 50+ years old 0.010614 13 Females 13 - 49 years old 0.009540 12Page 20 of 47

**5 .**  **4 .2**  **Chronic Dietary and Drinking Water Analysis** A conservative chronic dietary exposure anal ys is was performed for the general U.S. population and various population subgroups. Tolerance level residues and 100 crop treated assumptions were used. DEEM default and empirical processing factors were used to modify the tolerance values. The chronic anal ys is incorporated the 10 year average surface drinking water estimate from application of acibenzolar - S - methyl to cucurbit vegetables. The population subgroup s females 13 - 49 and young children had a more p rotective chronic population adjusted dose (cPAD, 0.082 mg/kg/day) than the general U.S. population and all other population subgroups (0.25 mg/kg/day). Chronic dietary risk estimates are not of concern for general population or other population subgroups . The subgroup s with the highest risk estimate w ere children 1 - 2 and 3 - 5 years old with a cPAD of 11 %. The cPAD for the general U.S. population was 2%. **Table 1 0**  **: Summary of Chronic Dietary Exposure and Risk to Fem**  **ale and Child Population for**  **Acibenzol ar**  **- S**  **- Methyl**  **Population Subgroup c**  **PAD (m g/kg/day)**  **Chronic Exposure (mg/kg/day)**  **% cPAD** Females 13 - 49 years old 0.082 0.003646 4 All Infants (< 1 year old) 0.003808 5 Children 1 - 2 years old 0.009207 11 Children 3 - 5 years old 0.008706 11 Children 6 - 12 years old 0.008706 7 **Table 1 2**  **: Summary of Chronic Dietary Exposure and Risk to Adults 50+ and Adult Males (13+)**  **Acibenzolar -**  **S -**  **Methyl Population Subgroup**  **cPAD (m g/kg/day)**  **Chronic Exposure (mg/kg/day)**  **% cPAD** General U.S. Population 0.25 0.004366 5 Males 13 - 19 0.003893 2 Males 20+ 0.003654 2 Adults 50+ years old 0.003852 2 **6 .0**  **RESIDENTIAL EXPOSURE AND RISK ASSESSMENT** A new residential exposure assessment is not required for the proposed new use of acibenzolar - S - methyl on low growing berries because there are no homeowner uses and/or application s to residential areas associated with this use . **6.1 Residential Bystander Postapplication Inhalation Exposure**  Based on the Agency 's current practices, a quantitative residential b ys tander postapplication inhalation exposure assessment was not performed for acibenzolar - S - methyl at this time. However, volatilization of pesticides may be a potential source of postap plication inhalation exposure to individuals nearb y to pesticide applications. The Agency sought expert advice and input on issues related to volatilization of pesticides from its Federal Insecticide, Fungicide, and Rodenticide Act Scientific Advisory Pan el (SAP) in December 2009. The Agency received the SAP’s final report on March 2, 2010Page 21 of 47

(http://www.epa.gov/scipoly /SAP/meetings/2009/120109meeting.html ). The Agency is in the process of evaluating the SAP report and may , as appropriate, develop policies and procedures, to identify

the need for and, subsequently , the way to incorporate postapplication inhalation exposure into the Agency's risk assessments. If new policies or procedures are put into place, the Agency may revisit the need for a quantitative postapplication inhalation exposure assessment for acibenzolar - S - methyl. **6.2 Spray Drift**  Spray drift is alway s a potential source of exposure to residents nearby to spraying operations. This is particularly the case with aerial application, but, to a l esser extent, could also be a potential source of exposure from the ground applicatio n method employed for acibenzolar S - methyl . The Agency

has been working with the Spray Drift Task Force, EPA Regional Offices and State Lead Agencies for pesticide regulation and other parties to develop the best spray drift management practices (see the Agency ’s Spray Drift website for more information at http://www.epa.gov/opp00001/factsheets/spraydrift.htm). On a chemical by chemical basis, the Agency is now requiring interim mitigation measures for aerial applications that must be placed on product labels/labeling. The Agency has completed its evaluation of the new database submitted by

the Spray

Drift Task Force, a membership of U.S. pesticide registrants, and is dev eloping a policy

on how to appropriatel y apply the data and the AgDRIFT computer model to its risk assessments for pesticides applied b y air, orchard airblast and ground hydraulic methods. After the policy

is in place, the Agency

may impose further refine ments in spray drift management practices to reduce off - target drift with specific products with significant risks associated with drift. Although a quantitative residential post - application inhalation exposure assessment was not performed as a result of pesticide drift from neighboring treated agricultural fields, an inhalation exposure assessment was performed for flaggers. This exposure scenario is representative of a worse case inhalation (drift) exposure and may be considered protective of most outdo or agricultural and commercial post - application inhalation exposure scenarios . **7 .0**  **AGGREGATE EXPOSURE AND RISK ASSESSMENT** 

In accordance with the FQPA, when there are potential residential exposures to a pesticide, aggregate risk assessment must consider exposures from three major routes: oral, dermal, and inhalation. There are three sources for these types of exposures: food, drinking water, and reside ntial uses. In an aggregate assessment, exposures from relevant sources are added together and compared to quantitative estimates of hazard (e.g., a NOAEL or PAD), or the risks themselves can be aggregated. When aggregating exposures and risks from vario us sources, HED considers both the route and duration of exposure. Acute and chronic aggregate exposures include food plus drinking water exposures. Acute and chronic aggregate risks are not of concern.Page 22 of 47

**7 .1**  **Short -**  **Term Aggregate Risk** To assess short -- term aggregate risk likely to result from the new and existing uses, HED combined average food and water exposure values with estimates of residential exposure for adult handlers and with residential exposure for adult and child post - application activit ies Tables 1 1 and 1 2 . Exposure for adult and child golfers was used to aggregate adult post - application dermal exposure with dietary and drinking water exposure. The aggregate exposure assessment for children combines dermal and incidental oral post - application exposure with food and water exposure. **Ta ble**  **11 . Short**  **- Term** **Aggregate Risk Calculations -**  **Residential Handler LOC =100**  **Population Subgroup Average**  **Food & Water Exposure mg/kg/day**  **Residential Exposure**  **1 mg/kg/day**  **Aggregate MOE (food and**  **residential) 2**  Females 13 - 49 y

rs 0.0038 0.00 73 700 1 Residential Exposure = [Dermal exposure + Inhalation Exposure]. 2 Aggregate MOE = [NOAEL ( 8.2 mg/kg/day  (Avg Food & Water Exposure + Residential Handler Exposure)] **Table 12**  **. Short -**  **Term Aggregate Risk Calculations**  **- Residential Post**  **- Application Expo**  **sure LOC =100 Population Subgroup**  **Average Food & Water Exposure**  **mg/kg/day Residential**  **Exposure 1**  **mg/kg/day Aggregate MOE**  **(food and residential)**  **2** Females 13 - 49 y

rs 0.0038 0.00 135 1600 Children 6 - 12 y

rs 0.0059 0.00 194 1 000 Children 1 - 2 y

rs 0.0093 0.0009 800 Children 3 - 5 y

rs 0.0088 850 1 Residential Exposure = [Dermal exposure + Inhalation Exposure]. 2 Aggregate MOE = [NOAEL (8.2 mg/kg/day  (Avg Food & Water Exposure + Residential Exposure)] Adult residential post application exposure from golfer scenario. Child residential exposure includes incidental oral and dermal exposures. **8.0 CUMULATIVE RISK**  Section 408(b)(2)(D)(v) of FFDCA requires that, when considering whether to establish, modify , or revoke a tolerance, the Agency consider “available information” concerning the cumulative effects of a particular pesticide's residues and “other substances that have a common mechanism of toxicity .” EPA does not have, at this time, available data to det ermine whether acibenzolar - S methyl has a common mechanism of toxicity with other substances. Unlike other pesticides for which EPA has followed a cumulative risk approach based on a common mechanism of toxicity , EPA has not made a common mechanism of tox icity finding as to acibenzolar - S - methyl and an y other substances and, acibenzolar - S - methyl does not appear to produce a toxic metabolite produced b y other substances which have tolerances in the U. S. For the purposes of this tolerance reassessment actio n, therefore, EPA has not assumed that acibenzolar - S - methyl has a common mechanism of toxicity with other substances. For information regarding EPA’s efforts to determine which chemicals have a common mechanism of toxicity and to evaluate the cumulative e ffects of such chemicals, see the policy statements released by EPA’s OPP concerning common mechanism determinations and procedures for cumulating effects fromPage 23 of 47

substances found to have a common mechanism on EPA’s website at http://www.epa.gov/fedrgstr/EPA_PEST/2002/January /Day _16/ . **9 .0**  **OCCUPATIONAL EXPOSURE /** **RISK CHARACTERIZATION**  **9 .1**  **Exposure Scenarios** O ccupational handler and post - application exposure scenarios were assessed for th e risk assessment of the proposed new use s on low growing berries . Based on the product labels and information provided b y the registrant, short - and intermediate - term exposure is assessed for occupational handlers and post - application activities. Dermal and inhalation exposures to workers are aggregated for acibenzolar - S - methyl because the toxicity endpoints for these exposure routes are based on common toxicological effects. **9 .**  **2 Handler**  **Exposure** The term “handler” applies to individuals who mix, load, and apply the pesticide product. The following handler exposure scenarios were assessed for proposed new acibenzolar - S - methyl uses. **9.2.1 Handler Exposure Scenarios** 

- Open mixing/loading (dry flowable) for aerial application;

- Open mixing/loading (dry flowable) for groundboom application;

- Applying spray s via aerial applications;

- Applying spray s via groundboom applications

- Flagging for aerial applications **9.2.2 Handler**  **Exposure Data** No chemical - specific handler exposure data were submitted in support of this registration. It is the policy of the HED to use surrogate data from the US Environmental Protection Agency , Office of Pesticide Programs , Occupational Pesticide Handler Unit Exposure Surrogate Reference Table, June 21, 2011, which summarizes and extracts surrogate data from the Pesticide Handlers Exposure Database Version 1.1 (PHED 1.1), the Agricultural Handler Exposure Task Force (AHETF) database, and the Outdoor Residential E xposure Task Force (ORETF) database. Some of these data, such as the industry task force data, are compensatory , subject to the data protection provisions of FIFRA. HED policies on use of surrogate data, including their sources, are presented on the inter net at http://www.epa.gov/pesticides/science/handler - exposure - data.html . The “Occupational Pesticide Handler Unit Exposure Surrogate Reference Table” may be found at http://www .epa.gov/pesticides/science/handler - exposure - table.pdf . **9.2. 3**  **Handler Exposure Assumptions**  The following standard exposure assumptions were used in estimating risks to workers from exposure to acibenzolar - S - methyl for the proposed new use on low growing berries . ThePage 24 of 47

daily areas treated were defined for each handler scenario b y determining the amount that can be reasonable treated in a single day . When possible, the assumptions for daily areas treated are taken from HED’s ExpoSAC Policy #9 (: “Standard Values for Daily Acres Treated in Agriculture”, which was revised on July 5, 2000.

• Average body weight of an adult handler is 6 0 kg.

• Exposure duration is short - term and intermediate - term for all workers assessed.

• Maximum application rates as determined b y l abel review were used for all ty pes and methods of application. The maximum single application rate for low growing berries is 0.02 3 lbs ai per acre.

• Estimated dermal absorption is 4 4 %

• SOP daily

volumes handled and/or area treated used for the scenarios assessed are:

- 80 acres treated per day for groundboom mix er/loader and applicator

- 350 acres treated per day for applications with fixed wing aircraft and flagging **9.2. 4**  **Handler Exposure and Risk**  **Estimates** A target LOC or MOE of 100 is considered adequate for inhalation and dermal exposure . All worker exposures are assessed as short - and intermediate - term based on label prescribed uses and expected exposure durations. Dermal and inhalation exposures to workers should be aggregated for acibenzolar - S - methyl because the same oral toxicity endpoint was selected for these exposure routes . Exposure and risk estimates indicate MOEs are not of concern (MOEs > 100) at the max imum use rate for occupational handler activities for the proposed new uses. A summary of occupational handler exposure and risk calculations, assumptions, and results is provided in Table 13. **Table 13**  **: Short/Intermediate -**  **Term Occupational Handler Exposure and Risk for Acibenzolar**  **- S**  **- Methyl**  **for Application to Low Grow**  **ing Berries Exposure**  **Scenario 1**  **App**

**Rate lb**  **a i/A**  **2 Acres** 

**Treat d**  **/ day**  **Unit Exposure 3**  **(ug/lb ai) Dose (mg/kg/Day)**  **4,5 MOEs**  **6**

**Dermal Inhal**  **Dermal Inhal**  **Dermal Inhal**  **Combined Derm & Inhal**  **Baseline Gloves**  **BL G**  **BL G**  **BL G** 

**Mi xer/Loader (dry flowable)**  Aerial 0.023 350 227 52 8.96 0.01 3 3 0.003 0 0.001 2 620 2700 6800 5 7 0 1800 Ground boom 80 0.003 0 0.000 6 0.000 3 2500 11000 27000 25 00 8000 **Applicator (dry flowable)** Aerial 0.023 350 0.005 Eng. C N/A 0.068 0.000 3 NA 9 E - 0 6 2 8 000 NA 90 0000 25000 NA Ground boom 80 78.6 16.1 0.0003 0.001 1 0.000 2 1E - 05 78 00 35000 7 9 0000 77 00 37 000 **Flagger (dry flowable)** Aerial 0.023 350 11 12 0.35 0.000 6 0.000 7 5E - 05 13 000 12 000 17 0000 1 2000 11 000 1 Use patterns are from the proposed labels 2 Application rates are based on maximum values based on proposed label. 3 Unit exposures for baseline (no personal protective equipment (PPE) and PPE (e.g., gloves) . Values are reported in the PHED Surrogate Exposure Guide dated August 1998 or are from data submitted by the Outdoor Residential Exposure Task Force dated May 2000.Page 25 of 47 4 Inhalation dose (mg/kg/day) = [unit exposure (ug/lb ai) * 0.001 m g/u g unit conversion * Inhalation absorption (100%) * Application rate (lb ai/acre or lb ai/gallon) * Daily area treated/amount handled (acres or gallons)] / Body weight ( 6 0 kg). 5 Dermal Dose (mg/kg/day) = [unit exposure ( mg/lb ai) * A pplication rate (lb ai/acre or lb ai /gallon) * Daily area treated/amount handled (acres or gallons) * Dermal Absorption Factor (44 %) ] / Body weight ( 6 0 kg). 6 Aggregate MOE = NOAEL (8.2 mkd)/Daily Inhalation Dose + Daily Dermal Dose **9.3 Post**  **Application Exposure** HED uses the term “post - a pplication” to describe those individuals who can be exposed to pesticides after entering areas previousl y treated with pesticides and performing certain tasks or activities . F or acibenzolar - S - methyl, post - application dermal exposure to field workers foll owing foliar applications is probable since acibenzolar - S - methyl is a fungicide that can be applied to all stages of growth. Since no post - application data were submitted in support of the registration action, dermal exposures during post - application acti vities were estimated using dermal transfer coefficients from the Science Advisory Council for Exposure Policy Number 3 (http://www.epa.gov/pesticides/science/exposac_policy 3.pdf ) . Based on the Agency 's current practices, a quantitative occupational post - application inhalation exposure assessment was not performed for acibenzolar - S - methyl at this time. However, there are multiple potential sources of post - application inhalation expo sure to individuals performing post - application activities in previously treated fields. These potential sources include volatilization of pesticides and re - suspension of dusts and/or particulates that contain pesticides. The Agency sought expert advice and input on issues related to volatilization of pesticides from its Federal Insecticide, Fungicide, and Rodenticide Act Scientific Advisory Panel (SAP) in December 2009. The Agency received the SAP’s final report on March 2, 2010 (http://www.epa.gov/scip oly /SAP/meetings/2009/120109meeting.html ). The Agency is in the process of evaluating the SAP report as well as available postapplication inhalation exposure data generated b y the Agricultural Reentry Task Force and may , as appropriate, develop policies and procedures, to identify the need for and, subsequently , the way to incorporate occupational post - application inhalation exposure into the Agency's risk assessments. If new policies or procedures are put into place, the Agency may revisit the need for a quantitative occupational post - application inhalation exposure assessment for acibenzolar - s - methyl. **9.3.1 Post Application Exposure Scenarios** 

• Handweeding • Scouting • Transplanting • Hand harvesting, scouting, and raking • Irrigating **9 .3.2**  **Post Application Exposure Assumptions** 

• Average body weight is 6 0 kg.

• T ransferrable residue is application rate times the fraction initially available (20%) . • Maximum transfer coefficient for post - application activities is 1900 cm 2 /hour ( irrigation) . • Exposure duration is 8 hours day .Page 26 of 47 • Exposure is assumed to occur on the day of application (day

0) . **9.3.3 Post**  **- Application Exposure and Risk Estimates**  A target LOC or MOE of 100 is considered adequate for dermal exposure . Exposure and risk estimates indicate MOEs are not of concern (MOEs > 100) at the maximum use rate for occupational post - application exposure activities for the proposed new uses . A summary of post application exposure and risk calculations , assumptions, and results is provided in Table 14. **Table 14 . Summary of Estimated Post**  **- application MOEs for Agricultural Crops**  **Crop Application** 

**Rate (lb ai/A)**  **1 DAT**  **2 DFR**  **3 (μg/cm** **2 )**  **TC 4** 

**(cm 2**  **/hr) Activity**  **4 Short**  **- /Int**  **** **Term MOE 5**  Lo w Growing Berries 0.02 3 0 0.05 2 70 Hand weeding 39 ,000 210 Scouting 13 ,000 230 Transplanting 12 ,000 1,100 Hand harvesting,

Scouting, Raking 2,5 00 1,900 Irrigation 1,4 00 1 Maximum application rate from proposed label 2 DAT = Days after treatment needed to reach the LOC of 100; DAT 0 = the day of treatment/ assumed to be approx. 12 hours. 3 DFR (μg/cm2 ) = dislodgeable foliar residues corresponding to DAT, based on 20% of application rate. 4 TC (cm2 /hr) = transfer coefficients and associated activities from ExpoSAC Policy Memo #003.1, 8/17/2000. 5 MOE = MOE on the corresponding DAT. MOE = NOAEL / Daily Dose. Daily Dose = [( DFR x TC x 4 0% Dermal absorption x 8 - hr Exposure Time)] / [(CF: 1000 μg/mg) x (60 - kg Body Weight) Short - /intermediate - term NOAEL = 8.2 mg/kg/day. The LOC is 100 **9.3. 4**  **Restricted Entry Interval** Acibenzolar - S - m eth ylhas been classified in Toxicity Category III for acute dermal and eye irritation and Category IV for primary

skin irritation. Per the Worker Protection Standard (WPS), a 12 - hr restricted entry interval (REI) is required for chemicals classified unde r Toxicity Category III/ IV. **10. REFERENCES**  Acibenzolar - S - Meth yl . Petition for the Establishment of Permanent Tolerances on Low Growing Berries (Subgroup 13 - 07G). Summary of Anal yt ical Chemistry

and Residue Data , T. Morton, D 386364 , 2/23/12 Acibenzolar - S - Meth yl

–Acute and Chronic Dietary and Drinking Water Exposure and Risk Assessment to support a Section 3 Request for Use of Acibenzolar - S - Methyl on Low Growing Berries Crop Subgroup 13 - 07G , T. Morton, D 38 7069, 2/23/12 Occupational Exposure A ssessment to Support the Request for Registration of Acibenzolar - S Methyl on Low Growing Berries (subgroup 13 - 07 G) , J.S. Miller, D 387841 , 2/23/12 Acibenzolar: Subchronic Neurotoxicity Study in Rats, K. Oo, D392694, 8/3/11Page 27 of 47

Absorption and Excretion of [U 14C ]Phenyl CGA 245704 in the Rat a fter Dermal Application , K. Oo, D392695 , 8/3/11 . Acibenzolar, Drinking Water Exposure Assessment

– R. Miller, D 353745 , 9 /2 /10Page 28 of 47

**APPENDI CES**  **A. TOXICOLOGY DATA SUMMARY**  **A .1**  **Guideline Data Requirements**  **Guideline No.**  **Study Type Technical**  **MRID No.**  **Required Submitted**  870.3100 870.3150 870.3200 870.3250 870.3465 Subchronic (Oral) Toxicity - Rodent ................................

Subchronic (Oral) Toxicity - Non - Rodent ................................ 21/28 - Day Dermal Toxicity ................................ 90 - Day Dermal Toxicity ................................ ................................ 90 - Day Inhalation Toxicity ................................ Y Y N N N Y

Y Y N N 44014230 44014228 44014232 42090013 46950310 870.3700a 870.3700b 870.3800 Prenatal Developmental Toxicity - Rodent ................................

Prenatal Developmental Toxicity - Non - Rodent ................................ Reproduction and Fertility Effects ................................ Y Y Y Y

Y Y 44014236 45089701 44014237 44014241 870.4100a 870.4100b 870.4200a 870.4200b 870.4300 Chronic (Oral) Toxicity - Rodent ................................

Chronic (Oral) Toxicity - Non - Rodent (Dog) ................................ Carcinogenicity - Rat.................. ................................ Carcinogenicity - Mouse ................................ ................................ Combined Chronic Toxicity /Carcinogenicity Y Y Y Y Y Y

Y Y Y Y 44014325 44014234 44014243 44014325 44014325 870.6100a 870.6100b 870.6200a 870.6200b 870.6300 870.7800 Neurotoxicity - Acute Delayed Neurotox. - Hen ................................

Neurotoxicity - Subchronic - Hen ................................ Neurotoxicity - Acute - Rat ................................ ................................ Neurotoxicity - Subchronic - Rat ................................ Developmental Neurotoxicity.................. Immunotoxicity............................. ................................ N N Y Y N Y N

N Y Y Y Y * ---

---

--- 45713601 46046401 48688301 * Studies have been submitted and are under reviewPage 29 of 47

**A .2**  **Toxic ity Profiles**  **Table 2**  **Acute Toxicity Profile Acibenzolar**  **- S**  **- methyl**  **Acibenzolar -**  **S -**  **methyl Guideline No./**  **Study Type MRID No.**  **Results Toxicity**  **Category** 870.1100 Acute oral toxicity 44014214 LD 50 >5000 mg/kg IV 870.1200 Acute dermal toxicity 44014216 LD 50 >2000 mg/kg IV 870.1300 Acute inhalation toxicity 44014219 LC 50 >5.022 mg/L IV 870.2400 Acute eye irritation 44014220 minimal III 870.2500 Acute dermal irritation 44014220 slight IV 870.2600 Skin sensitization 44014225 positive NA **Actigard 50WG** 870.1100 Acute oral toxicity 44014215 LD 50 >5000 mg/kg IV 870.1200 Acute dermal toxicity 44014217 LD 50 >2000 mg/kg IV 870.1300 Acute inhalation toxicity 44014218 LC 50 >2.79 mg/L IV 870.2400 Acute eye irritation 44014221 minimal III 870.2500 Acute dermal irritation 44014223 mo derate III 870.2600 Skin sensitization 44014224 negative NA **Table 3**  **Subchronic, Chronic and Other Toxicity Profile for Acibenzolar**  **- S**  **- me**  **thyl Guideline No.**  **Study Type MRID No. (year)**  **Classification Dose Levels**  **Results** 870.3100 90 - day oral toxicity rodents (rat) 44014230 (19 93 ) Acceptable/Guideline M : 0, 2.42, 24.6, 126, 516 mg/kg/day; F: 0, 2.64, 26.3, 131, 554 mg/kg/day NOAEL : 126 mg/kg/day Males or M); 131 mg/kg/day (Females or F) LOAEL : 516 mg/kg/day (M); 554 mg/kg/day females based decreased body weight, reduced food intake and efficiency and increased liver and spleen weights 870.31 50 90 - day oral toxicity non rodents ( dog ) 44014232 (1994) Acceptable/Guideline M & F: 0, 10, 50, 200 mg/kg/day (capsules) NOAEL : 50 mg/kg/day (M & F) LOAEL : 200 mg/kg/day ( F) based on regenerative hemolytic anemia evidenced by decreased erythroid parameters, pigmentation in the liver and spleen, splenic congestion, and by activation of medullary (bone marrow) and extramedullary (splenic) erythropoiesis. 870.3100 90 - day oral toxicity rodents ( mice ) 44014228 (1993) Acceptable/Guideline 0, 30.6, 152, and 624 mg/kg/day for males, and 0, 47.4, 220, and 803 mg/kg/day for females, respectively. NOAEL : 30 . 6 mg/kg/day ( M); 47.4 mg/kg/day (F) LOAEL : 152 mg/kg/day (M); 220 mg/kg/day females based decreased body weight and body weight gain, increased spleen weight and microscopic changes in the spleenPage 30 of 47

**Table 3**  **Subchronic, Chronic and Other Toxicity Profile for Acibenzolar**  **- S**  **- me**  **thyl Guideline No.**  **Study Type MRID No. (year)**  **Classification Dose Levels**  **Results** 870.3200 21/28 - day dermal toxicity (rat) 44014233 (1994) Acceptable/guideline M & F: 0, 10, 100, 1000 mg/kg/day NOAEL = 1000 mg/kg/day LOAEL = not identified 870.3700a Developmental Toxicity in rodents (rat) 44014236 (1994) Acceptable/guideline F: 0,10, 50, 200, 400 mg/kg/day Maternal NOAEL 200 mg/kg/day Developmental NOAEL 50 mg/kg/day Maternal LOAEL 400 mg/kg/day based on clinical signs of hemorrhagic perineal discharge Developmental LOAEL 200 mg/kg/day based on developmental malformations, anomalies, and skeletal variations 870.3700a Developmental Toxicity in rodents (rat) 4 5089701 (19 98 ) Acceptable/guideline of 0 , 10, 75 , 150, or 350 mg/kg/day Maternal NOAEL 350 mg/kg/day Developmental NOAEL 200 mg/kg/day Maternal LOAEL

– not established (>350) Developmental LOAEL 350 mg/kg/day based on increased incidences of rudimentary and long lumbar ribs 870.3700b Developmental Toxicity in non - rodents (rabbit) 44014237 (1994) Acceptable/guideline F: 0, 10, 50, 300, 600 mg/kg/day Materna l NOAEL = 50 mg/kg/day LOAEL = 300 mg/kg/day based on mortality, clinical signs of toxicity, decreased maternal body w eight and food consumption. Developmenta l NOAEL = 300 mg/kg/day LOAEL = 600 mg/kg/day based on a marginal increase in vertebral anomalies. 870. 6300 Developmental Neurotoxicity Toxicity in rodents (rat) 46046401 ( 2002 ) Acceptable/non - guideline 0/0, 8.2/15.5, 82.0/153.6, and 326.2/607.8 mg/kg/day [gestation/lactation]. Maternal NOAEL 326.2 mg/kg/day Developmental NOAEL 8.2 mg/kg/day Maternal LOAEL Not Observed Developmental LOAEL 82 based on changes in brain mo rphometrics in the cerebellum 870.3800 Reproduction and fertility effects in rats 4014241 (1995) Acceptable/guideline M & F: 0, 1 - 3, 11 - 31, 105 - 288, 223 - 604 mg/kg/day **Parental/System ic**  NOAEL = 11 - 31 mg/kg/day LOAEL = 105 - 288 mg/kg/day based on increased weights and hemosiderosis of the spleen. **Reproductive** NOAEL = 223 - 604 mg/kg/day LOAEL > 223 - 604 mg/kg/day based on no effects. **Offspring** NOAEL = 11 - 31 mg/kg/day LOAEL = 105 - 288 mg/kg/day based on reduced pup body weight gains and lower pup body weights during lactation.Page 31 of 47

**Table 3**  **Subchronic, Chronic and Other Toxicity Profile for Acibenzolar**  **- S**  **- me**  **thyl Guideline No.**  **Study Type MRID No. (year)**  **Classification Dose Levels**  **Results** 870.6200b Subchronic Neurotoxicity 45713601 (1997) acceptable/non guideline pending submission of positive control data for neuropathological and FOB assessments 0/0, 24/26, 126/143, and 575/628 mg/kg/day [M/F], respectively The study may be upgraded pendi ng submission of acceptable positive control data. NOAEL 126/143 mg/kg/day, M/F LOAEL 575/628 mg/kg/day, M/F based on decreased body weights, body weight gains, and food consumption .

870.4100 Chronic toxicity in dogs 44014234 (199 6 ) Acceptable/guideline M & F: 0, 5, 25, 200 mg/kg/day (capsules) NOAEL 25 mg/kg/day LOAEL 20 0 mg/kg/day based on hematological effects, hemosiderosis of the liver and spleen, extraniedullary hematopoiesis in the spleen, and increases in liver weight. 870.4300 Chronic toxicity/ carcinogenicity in rodents ( mice ) 44014325 (199 6 ) Acceptable/guideline M: 0, 1.14, 11.1, 237, 698 mg/kg/day F: 0, 1.14, 10.8, 234, 696 mg/kg/day NOAEL 11.1/10.8 mg/kg/day , M/F LOAEL 237/234 mg/kg/day based on mild hemolytic anemia and related microscopic changes (hemosiderosis) in the spleen, liver, and bone marrow

and extramedullary hematopoiesis in the spleen. 870.4200 Carcinogenicity study in rat 44014243 (199 6 ) Acceptable/guideline M: 0, 0.77, 7.77, 96.9, 312 mg/kg/day F: 0, 0.90, 9.08, 111, 388 mg/kg/day NOAEL 96.9/111 (M/F) LOAEL 312/388 / 191.0 mg/kg/d (M/F) based on decreased body weights, body weight gain and food efficiency, mild hemolytic anemia, and increased incidence of alveolar foam cells (females only). 870.5100 Bacterial reverse mutation assay (Ames test) 441014247 (1993) Acceptable/guideline 312.5, 625, 1250, 2500, 5000  g/plate Negative with and without S - 9 activation at 5000  g/plate and less. 870.5100 Bacterial reverse mutation assay (Ames test) Test Material: CGA - 362020 (is omer of acibenzolar - S methyl) 44537025 (1998) Acceptable/guideline 61.73 to 5000  g/plate (+S - 9) 30 - 86 to 2500  g/plate ( - S - 9) Positive in S. typhimurium strain TA1537 at 277.8  g/plate and higher in the absence of S - 9. Negative with S - 9 activation at 500 0  g/plate and less.Page 32 of 47

**Table 3**  **Subchronic, Chronic and Other Toxicity Profile for Acibenzolar**  **- S**  **- me**  **thyl Guideline No.**  **Study Type MRID No. (year)**  **Classification Dose Levels**  **Results** 870.5100 Bacterial reverse mutation assay (Ames test) Test Material: NOA - 419191 (by - product of acibenzolar - S methyl) 44537024 (1998) Acceptable/guideline 312.5 to 5000  g/plate (± S - 9) Negative with or without S - 9 activation at 5000  g/plate and less

870.5100 Bacterial reverse mutation assay (Ames test) Test Material: CGA - 323060 (plant metabolite of acibenzolar - S - methyl) 44537026 (1997) Acceptable/guideline 312.5 to 5000  g/plate (± S - 9) Negative with or without S - 9 activation at 5000  g/plate and less

870.5300 In vitro mammalian gene mutation assay 44014246 (1993) Acceptable/guideline 3.70 to 100  g/ml ( - S - 9), 37.04 to 1000  g/ml (+S - 9) Negative with S - 9 activation up to 1000  g/ml. Negative without S - 9 activation up to 100  g/ml. Com pound tested to cytotoxic concentrations. 870.5375 In vitro mammalian chromosome aberration (CHO cells) 44014245 (1993) Acceptable/guideline 7.5 to 60  g/ml ( - S - 9 and +S 9) Suggestive of clastogenicity i n the absence of S - 9 activation at 30 and 60  g/mL at the 18 - hour cell harvest time; effect observed only in the presence of cytotoxicity. Increase in polyploid cells at 30 and 60  g/mL at the 42 hour harvest time both with and without S 9. Evidence of c ell cycle arresting activity at G 2 . 870.5395 Mammalian erythrocyte micronucleus test 44014244 (1993) Acceptable/guideline 1000, 2000, 4000 mg/kg (oral gavage) Negative at 16, 24, and 48, hour sacrifices. 870.5550 UDS in primary rat hepatocytes 44014248 (1993) Acceptable/guideline 9.77 to 500  g/ml Negative at 500  g/ml and less. 870.7485 Metabolism and pharmacokinetics rats 44014250 (1995) Acceptable/guideline 0.5, 100 mg/kg Following oral treatment of rats, acibenzolar - S - methyl was rapidly and nearly completely (>90% of administered dose) absorbed from the gastrointestinal tract into the general circulation. The majority (88 - 95%) of the administered dose was excreted in the urine within the first 48 hours. The majo r metabolite (79 - 92%) in the urine was the carboxylic acid derivative of the parent. 870.7600 Dermal penetration NA NA Special studies : 28 - Day dietary rats 44014227 (1992) Acceptable/nonguideline M: 0, 45.9, 403, 1070 mg/kg/day; F: 0, 44.8, 376, 1000 mg/kg/day NOAEL = M: 403 mg/kg/day; F: 376 mg/kg/day LOAEL = M: 1070 mg/kg/day; F: 1000 mg/kg/day based on decreased mean body weights, decreased liver weights, altered hematology parameters accompanied by increased spleen weights.Page 33 of 47

**Table 3**  **Subchronic, Chronic and Other Toxicity Profile for Acibenzolar**  **- S**  **- me**  **thyl Guideline No.**  **Study Type MRID No. (year)**  **Classification Dose Levels**  **Results** 28 - Day oral gav age rats 44014229 (1993) Acceptable/nonguideline 0, 10, 100, 800 mg/kg/day NOAEL = 100 mg/kg/day LOAEL = 800 mg/kg/day based on decreased body weights, and decreased hemoglobin - related parameters accompanied by hemosiderosis of the spleen, increased liver and spleen weights, and decreased thymus weights. 28 - Day oral capsule dogs 44014231 (1994)

Acceptable/nonguideline 0, 50, 250, 500 mg/kg/day NOAEL = 50 mg/kg/day LOAEL = 250 mg/kg/day based on decreased body weight, decrea sed hemoglobin - related parameters, hepatic and splenic hemosiderosis. 90 - Day Dietary mice 44014228 (1993) Acceptable/nonguideline M: 0, 30.6, 152, 624 mg/kg/day; F: 0, 47.4, 220, 803 mg/kg/day NOAEL = M: 30.6 mg/kg/day; F: 47.4 mg/kg/day LOAEL = M: 152 mg/kg/day; F: 220 mg/kg/day based on decreased mean body weights and body weight gain in males, increased spleen weights and splenic histopathology in both sexes. 90 - Day Dietary dog 44014232 (1994) Acceptable/nonguideline 0, 10, 50, or 20 0 mg/kg/day NOAEL = 50 mg/kg/day LOAEL = 200 mg/kg/day in male and female dogs, based on regenerative hemolytic anemia evidenced by decreased erythroid parameters, pigmentation in the liver and spleen, splenic congestion, and by activation of medullary (b one marrow) and extramedullary (splenic) erythropoiesis. 90 - Day Dietary rat 440142 30 (1993) Acceptable/nonguideline males: 0,2.42, 24.6, 126, or 516 mg/kg/day; females: 0,2.64,26.3, 131, or 554 mg/kg/day) NOAEL = 126 mg/kg/day in males and 131 mg/kg/day in females LOAEL = 516 mg/kg/day males and 554 mg/kg/day females based on decreased mean body weights, reduced food intake, reduced food efficiency, and increased liver and spleen weights with correlates for the liver and spleen of glycogen deposition and hemosiderosis, respectively. Special Developmental toxicity rats 44014238 (1994) Acceptable/nonguideline 300 mg/kg/day, GD 6 - 15, 6 - 7, 8 - 9, 10 - 11, 12 - 13, or 14 - 15 Maternal and developmental NOAELS and LOAELS could not be identified by this protocol. The most pronounced maternal and developmental toxicity occurred w hen dams were treated on GD 6 - 15. Special Developmental toxicity rats 44014239 (1994) Acceptable/nonguideline 400 mg/kg/day, GD 6 - 7, 8 - 9, 10 - 11, 12 - 13, or 14 - 15 Maternal and developmental NOAELS and LOAELS could not be identified by this protocol. The most pronounced maternal and developmental toxicity occurred w hen dams were treated on GD 6 - 7 and 8 - 9. Dermal developmental toxicity rats 44014240 (1994) Acceptable/nonguideline 0, 10, 100, 500 mg/kg/day, GD 6 - 15 Maternal NOAEL  500 mg/kg/day LOAEL >500 mg/kg/day based on no effects. Developmental NOAEL  500 mg/kg/day LOAEL >500 mg/kg/day based on no effects . Range - finding 1 - generation reproduction rats 44681301 (1993) Acceptable/nonguideline 0, 199 - 209, 382 - 410, 700 - 728 mg/kg/day Parental/Systemic NOAEL = 209 mg/kg/day LOAEL = 410 mg/kg/day based on decreased body weight gain and food consumption in females. Reproductive NOAEL = 410 mg/kg/day LOAEL = 728 mg/kg/day based on total resorptions in all dams. Offspring NOAEL = 209 mg/kg/day LOAEL = 410 mg/kg/day based on reduced pup body weight gains and lower pup body weights during lactation.Page 34 of 47

**A.3 Toxicological Endpoints**  **A .3.1**  **Acute Reference Dose (aRfD) -**  **General Population** Selected Study : Developmental Neurotoxicity in R ats (MRID 46046401 ). See Section 3.2. 3 Dose and Endpoint for Establishing an aRfD : 8.2 mg/kg/day based on an offspring NOAEL of 8.2 mg/kg/day and an offspring LOAEL of 82 mg/kg/day based on changes in b rain morphometrics in the cerebellum. Uncertaint y Factor (UF ): 100

– 10 x for interspecies extrapolation and 10x for intraspecies variation. Comments about Study

/Endpoint : The developmental neurological effects of concern are presumed to occur after a single exposure and are relevant to females aged 13 - 50 years old since they occur in utero and to young children based on post - natal effects. [Note: In the previous risk assessment (D361893, 3/3/09) only the subpopulation females 13 - 50 ye ars old was assessed for acute dietary exposure based on in utero effects. However, based on a reevaluation of the DNT study regarding the potential for post - natal toxicity , the acute dietary anal ys is has been expanded to include children for this assessm ent.]

**A .3.**  **2 Chronic**  **Reference Dose ( c**  **RfD) -**  **Females age 13 -**  **49 and Young Children**  Selected Study : Developmental Neurotoxicity in R ats (MRID 46046401 ). See Section 3. 2 . 3 Dose and Endpoint for Establishing a cRfD: 8.2 mg/kg/day based on an offspring NOAEL of 8.2 mg/kg/day and an offspring LOAEL of 82 mg/kg/day based on changes in brain morphometrics in the cerebellum. Uncertaint y Factor (UF ): 100

– 10 x for interspecies extrapolation and 10x for intraspecies variation. Comments about Study

/Endpoint : The effects observed in this study can occur after single or multiple doses. Selection of a chronic endpoint from this s tudy ensures that the chronic exposure assessment is protective of f emales of reproductive age and their offspring. **Acute Rf**  **D = 8.2**  **mg/kg (NOAEL**  **) = 0.**  **0.082 mg/kg/day**  **(General Population) 1**  **00 (UF)** Page 35 of 47

[Note: In the previous risk assessment (D361893, 3/3/09) only the subpopulation females 13 - 50 years old was assessed for chronic dietary exposure based on in utero effects observed in the DNT study . However, based on a reevaluation of the DNT study regarding the potential for post - natal toxic ity , the chronic dietary anal ys is the chronic dietary exposure anal ys is has been revised to include an assessment of dietary risks to children using the NOAELfrom the DNT study .]

**A .3.**  **3 Chronic**  **Reference Dose (RfD) –**  **Adult Males and Females 50+ years** Selected Stud ies : Co - critical studies Chronic Feeding Study in Dogs (MRID 44014234 ); Oncogenicit y Study Mouse (MRID 44014235) ; Chronic/Oncogenicit y Study Rat (MRID 44014243) ; Reproductive Study Rat ( 44014241 ) Chronic Dog Study - In a chronic oral toxicity study acibenzolar - S - methyl was administered to 4 beagle dogs per sex per dose level in gelatin capsules at levels o f 0, 5, 25, or 200 mg/kg/day

for 52 weeks. No biologically significant toxic effects were observed in either males or females receiving 25 mg/kg/day or less of the test material. At 200 mg/kg/day , there was an indication of hemoly tic anemia in dogs of both sexes i.e., effects consistent with prior or current hemoly

tic anemia including hematological effects, hemosiderosis of the liver and spleen, extraniedullary

hematopoiesis in the spleen, and increases in liver weight. Significant treatment related chang es were found in clinical chemistry in the dogs fed the test, substance at 200 mg/kg/day , ( increase in trigl yc erides in both sexes, increase of cholesterol in females at week 13, decrease in total protein in males at weeks 26 and 52, and decrease in albumi n in both sexes at week 13. These clinical chemistry changes are suggestive of possible nutritional deficiencies. The LOAEL is 200 mg/kg/day in both sexes based on effects consistent with prior or current hemoly tic anemia . The NOAEL is 25 mg/kg/day . O ncogenicit y Mouse Study - In an oncogenicit y study acibenzolar - S - methyl w as administered in the diet to groups of 60 male and 60 female Tif:MAGf (SPF) mice at concentrations of 0, 10, 100, 2000, or 6000 ppm ( 1.14, 11.11, 237, and 698 mg/kg/day for males and 1.14, 10.8, 234, and 696 mg/kg/day for females ) for 18 months. Ten mice per group were reserved for measurements of hematology parameters after 12 and 18 months of treatment. Treatment did not result in a significant decrease in survival. The overal l food efficiency was decreased in females b y 22% at 6000 ppm . The red blood cell count, hemoglobin, and hematocrit at 53 and 79 weeks were decreased in females at 2000 ppm and 6000 ppm . M ean corpuscular volume was decreased in high dose males at 79 week s . M ean corpuscular hemoglobin concentration distribution increased at 6000 ppm . Reticulocy te counts in high - dose females was significantly increased. The hematology data are consistent with hemoly tic anemia especiall y in females at 2000 and 6000 ppm. Macroscopic and microscopic findings consistent

**Chronic Rf** 

**D =**  **(NOAEL) 8.2**  **mg/kg =**  **0.082 (**  **F emales 13**  **- 49**  **/Young Children )**  **(UF) 100**Page 36 of 47

with a mild hemoly

tic anemia in both sexes included increased incidences of enlarged spleen , increased incidences of hemosiderosis in liver , spleen , and bone marrow . I ncidence of extram edullary hematopoiesis in the spleen and pancreatic exocrine hy perplasia was increased in males at 2000 and 6000 ppm. In cidences of alveolar foam cells in the lungs were increased at 6000 ppm in both sexes . The LOAEL is 2000 ppm (237 /234 mg/kg/day males / females), based on mil d hemoly

tic anemia and related microscopic changes (hemosiderosis) in the spleen, liver, and bone marrow and extramedullary hematopoiesis in the spleen. The NOAELwas 100 ppm (11.1 mg/kg/day

males; 10.8 mg/kg/day

females). Chronic/Oncogenicit y Rat Study - In a chronic toxicity /oncogenicity

study, acibenzolar S - methyl was administered in the diet to groups of 80 male and 80 female (Tif:RAIf (SPF)) Sprague - Dawley

derived rats at concentrations of 0, 20, 200, 2500, or 7500 ppm ( 0, 0.77, 7.77, 96.9, and 312 mg/kg/day male and 0, 0.90, 9.08, 111, and 388 mg/kg/day female ) . Ten animals per sex per dose were sacrificed at 52 weeks for interim evaluation; the remaining animals were used for hematologic and clinical chemistry evaluation (20 per sex per group) and evaluation of carcinogenic potential (50 animals per sex per group). No treatment - related effects were observed at 20 , 200 or 2500 ppm for an y parameter. Treatment related effects on b ody weight, weight gain, food consumption, and food efficiency were o bserved in both sexes at 7500 ppm . Other treatment - related effects at 7500 ppm included a slight hemol yt ic anemia in both sexes as evidenced b y decreases in ery throcy te count, increased mean corpuscular volume, elevated reticulocy te counts, increased serum bilirubin, increased severit y of hemosiderin deposition in the spleen, and increased incidence of hemosiderin deposition in Kupifer cells of the liver. Hemosiderin deposition in the spleen was accompanied b y a statistically significant increase in splee n weight. There was also a statistically significant increase in the incidence of foam cells in the lungs of females fed the 7500 - ppm diet. The LOAEL was 7500 ppm (312 /388 mg/kg/day males/ females) based on decreased body weights, body weight gain and foo d efficiency , mild hemoly tic anemia, and increased incidence of alveolar foam cells (females only). The NOAEL was 2500 ppm (96.9 mg/kg/day males and 111 mg/kg/day females). Reproducti ve Study

in Rats - See Section 3.2. 2 Dose and Endpoint for Establishi ng cRfD : 25 mg/kg/day based on a NOAELof 25 mg/kg/day

from the chronic dog study and a LOAELof 105 mg/kg/day from the rat reproduction study based on hemoly

tic anemia with compensatory response. Uncertaint y Factor(s) : 100

– 10x for interspecies extrapolation and 10x for intraspecies variation. Comments about Study

/Endpoint/Uncertaint y Factor : The NOAEL of 25 mg/kg/day , taken from the chronic dog study, is based on the totality of the database with a L OAEL of 105 mg/kg/day

(the lowest LOAEL is from the reproduction study) based on hemoly tic anemia with compensatory

response. The NOAEL and LOAEL recommended for use in the risk assessment come from different studies conducted in different species . Ty picall y, a combination of NOA ELs/LOAELs from studies conducted in different species is not recommended. However,Page 37 of 47

in the case of acibenzolar - S - methyl the same endpoint (hemoly tic anemia) is observed across species at a relativel y close range of LOAELs (105 - 312 mg/kg/day). Therefore, the ToxSAC concluded that, in this instance, it would be appropriate to base the point of departure for the risk assessment on the NOAEL from the dog chronic toxicity study using the LOAEL from the reproduction study in rats for the basis of the endpoint (hemoly tic anemia).

**A .**  **3 .**  **4 Incidental Oral Exposure (**  **S hort**  **- and**  **I ntermediate**  **- Term)**  Selected Study : Developmental Neurotoxicity in R ats (MRID 44014236 ). See Section 3.2. 3 Dose and Endpoint : 8.2 mg/kg/day based on an offspring NOAEL of 8.2 mg/kg/day and an offspring LOAEL of 82 mg/kg/day based on changes in brain morphometrics in the cerebellum. Uncertaint y Factor (UF ): 100

– 10 x for interspecies extrapolation and 10x for intraspecies variation. Comments about Study

/Endpoint : This endpoint is considered appropriate based on the post - natal component of the DNT. **A .**  **3 .**  **5 Dermal Absorption**  A dermal absorption factor (DAF) is applied when dermal exposure endpoints are selected from oral toxicity studies. The dermal factor converts the oral dose to an equivalent dermal dose for the risk assessment. A DAF of 45 % was selected for use in risk a ssessment based on a dermal absorption study in rats. **A .**  **3 .**  **6 Dermal Exposure (**  **Short and Intermediate -**  **Term )**  Selected Study : Developmental Neurotoxicity in R ats (MRID 44014236 ). See Section 3.2. 3 Dose and Endpoint : 8.2 mg/kg/day based on an offspring NOAEL of 8.2 mg/kg/day and an offspring LOAEL of 82 mg/kg/day based on changes in brain morphometrics in the cerebellum. Uncertaint y Factor (UF ): 100

– 10 x for interspecies extrapolation and 10x for intraspecies variation. **Chronic RfD = 25**  **mg/kg/day (NOAEL**  **) = 0.**  **25 mg/kg/day** 

**(Adult Male) 1**  **00 (UF)** Page 38 of 47

Comments about Study

/Endpoint : Two acceptable dermal toxicity studies, including a dermal developmental study were available. However s ince neurological effects observed in the oral developmental neurotoxicity study are not measured in the dermal developmental study , the findings from the dermal study may not be protective of these neurological effects . Since an oral NOAEL was selected for dermal risk assessment , a dermal absorption factor of 4 5 % should be used . This factor is based on a dermal absorption study in rats. **A .3.**  **7 Inhalation Exposure**  **( Short and Intermediate**  **- Term** **)** Selected Study : Developmental Neurotoxicity in R ats (MRID 44014236 ). See Section 3.2. 3 Dose and Endpoint : 8.2 mg/kg/day based on an offspring NOAEL of 8.2 mg/kg/day and an offspring LOAE L of 82 mg/kg/day based on changes in brain morphometrics in the cerebellum. Uncertaint y Factor (UF ): 100

– 10 x for interspecies extrapolation and 10x for intraspecies variation. Comments about Study

/Endpoint : Since an oral NOAEL was selected for inhalation risk assessment, an inhalation absorption factor of 100% should be usedPage 39 of 47

**A.4 EXECUTIVE SUMMAR** **IES 870.3700 Developmental Toxicity**  In a non - guideline developmental toxicity study (MR ID 44014238), twelve presumed pregnant Tif:RAT f(SPF) (h ybrids of RTT/ 1 x RII/2) rats per group were administered CGA 245704 Technical (97.9%; Batch No. P.303011) in a 0.5% aqueous solution of sodium carbox ym eth yl cellulose by gavage at a dose of 300 mg/kg/d ay . Groups 2 - 7 were administered test article on gestation day s (GD) 6 - 15,6 - 7, 8 - 9, 10 - 11 , 12 - 13, or 14 - 15, respectivel y. Group 1 consisted of twelve animals which served as controls and were treated with 0.5% sodium carbox ym eth yl cellulose on GD 6 - 15. On GD 21, dams were sacrificed, subjected to gross necrops y, and all fetuses examined externally . Visceral and skeletal examinations were not performed. All animals survived until scheduled sacrifice on GD 21. Bloody , vaginal discharge was observed in 8/12 d ams in Group 2, 3/ 1 2 dams in Group 3, 5/12 dams in Group 4, 4/12 dams in Group 5, and in 5/12 dams in Group 6. This f in ding was observed for one to four day s starting on GD 13 and was not observed in controls or in animals in Group 7. No other treatment - re lated clinical signs of toxicity

were observed. No statistically significant differences in absolute body weights or body weight changes occurred at an y time between the treated groups and the control group. Body weight gain by Group 2 was 80% of the control level during the treatment interval (GD 6 - 1 6), with the greatest reduction in weight gain (77% of controls) during GD 11 - 16. Food consumption was not affected. There were no statistically significant differences between control and treatment groups for number of corpora lutea, number of implantation sites, live fetuses/dam, pre - and post - implantation loss, or fetal sex ratios. Fetal body weights from the Group 2 dams were significantly

(p <0.01) less than the controls. Although not statistically signifi cant, Group 2 dams also had fewer live fetuses per litter (10.9 vs 14.8 for controls) and a greater post implantation loss (30.1% vs 2.9% for controls) from earl y resorptions due to one dam with whole liner resorption ( 1 7/17) and 2 additional dams with 13/ 16 and 8/9 resorptions/number of implantation sites. The overall incidence rates for litters containing fetuses with major external malformations in Groups 1 - 7 were 0/12, 1/10, 0/ 10 , 1/ 11 , 0/ 11 , 0/10, and 0/12, respectivel y. In one Group 2 litter, generali zed edema was observed in 5/14 fetuses. In addition, two of the five also had a position anomaly of the hindlimb and one of the two also had gastroschisis. These five fetuses also had greatl y reduced body weights. Generalized edema was also seen in one fet us from Group 4. Maternal and developmental toxicity NOAELs and LOAELs could not be identified by

this protocol, which investigated the critical period of organogenesis. The most pronounced maternal and developmental toxicity occurred when dams were treat ed on GD 6 - 15, inclusive at a dose level of 300 mg/kg/day . This study is classified as Acceptable/non - guideline as a developmental toxicity study (83 - 3a) in rats. In a non - guideline developmental toxicity study (MR ID 44014239), eight presumed pregnant Tif : RAT f (SPF) (h ybrids of RII/1 x R II 2) rats per group were administered CGA 245704 Technical (97.9%; Batch No. P.303011) by gavage at a dose of 400 mg/kg/day . Groups 2 - 6 were administered test article on gestation day s (GD) 6 - 7, 8 - 9, 10 - 11, 12 - 13, or 14 - 1 5, respectively . Group 1 consisted of eight animals which served as controls and were treated with 0.5% carbox ym eth yl celiulose (vehicle) on GD 6 - 15. On GD 21, dams were sacrificed, subjected toPage 40 of 47

gross necrops y, and all fetuses examined externally. Visceral and skeletal examinations were not performed. One Group 2 dam was found dead on GD 20. Prior to death, this animal had bloody perineal discharge and necrops y findings including mottled kidney and th ymus. One Group 4 dam delivered on GD 20 and was sacrific ed. All other animals survived until scheduled sacrifice on GD 21. Bloody, perineal discharge was observed in 4/8 dams in Group 3. This f in ding was observed for three to four day s starting on GD 16. The same 4 dams were later observed to have total lifter resorptions. No other treatment - related clinical signs of toxicity were observed. No statistically

significant differences in body weights, body weight gains, or food consumption occurred at an y time between the treated groups and the control group. There were no differences between the control group and Groups 4, 5, and 6 for number of corpora lutea, number of implantation sites, live fetuses/dam, pre - and post - implantation loss, fetal body weights, or fetal sex ratios. Group 2 had a slightly greater numb er of earl y resorptions/dam (2.7 vs 0.9 for controls) and an increased postimplantation loss (18.7% vs 5.4% for controls). The four Group 3 dams observed with bloody discharge had complete litter resorption resulting in increased postimplantation loss (53. 4%). Group 3 also had an increase in early rcsorptions (8.0/dam vs 0.9/dam for controls) and decreased mean live fetuses/litter (8.5 vs 16.0 for controls). Another Group 3 dam had fused placenta between two fetuses. The overall incidence rates for liners containing fetuses with major external malformations in Groups 1 - 6 were 0/8, 0/6, 0/4, 1/7, 0/8, and 0/8, respectivel y. One Group 4 fetus had cleft palate and another fetus from the same lifter had a kinked tail. No treatment - related external m alformations /variations were observed in an y fetus from any group. Maternal and developmental toxicity NOAELs and LOAELs could not be identified by this protocol, which investigated the critical period of organogenesis. Pronounc ed maternal and developmental toxicity occurred when dams were treated on GD 6 - 7 and 8 - 9 at a dose level of 400 mg/kg/day . **870.7600 Dermal Toxicity Studies**  In a dermal developmental toxicity study (MRID 44014240 ) , 24 pregnant Tif: RA I f (SPF) rats (Sprague - Dawley

derived) per group were administered acibenzolar - S - methyl (97.9% a.i.) in aqueous carbox ym eth yl cellulose (0.5% w/w) in 0.1% aqueous Poly sorbate 80, by dermal application at doses of 0 (vehicle), 10, 100, or 500 mg/kg/day on gest ation day s (GD) 6 - 15, inclusive. Exposure duration was 6 hours per day . On GD 21, all dams were sacrificed and all fetuses were sexed, weighed, and examined for external malformations/variations, then sacrificed. Approximately one - half of each litter was placed in Bouin’s fixative for subsequent visceral examination and the remainder w ere prepared and stained for skeletal examination. All animals survived until scheduled sacrifice. No clinical signs of toxicity were observed at an y treatment level. Gest ational body weights, body

weight changes, and food consumption were not statistically

significantly

different for an y treatment level throughout the gestational period. Therefore, the maternal toxicity NOAEL is 500 mg/kg/day based on no adverse maternal e ffects at the highest administered dose. The maternal toxicity L OAEL was >500 mg/kg/day . No treatment - related effects were observed for gravid uterine weights, number of fetuses/litter, pre and post - implantation losses, numbers of corpora lutea/dam, number of implantations/dam, resorptions/dam, fetal body weights, or fetal sex ratios. No biologicall y relevant statistically significant differences in the incidence rates of any external, visceral, or skeletal malformations/Page 41 of 47

anomalies/variations were obser ved in the treated litters as compared to the controls. There were statistically

significantly

decreased incidences of absent ossification of the proximal phalanx of anterior digit - 2 in all treatment groups and poor ossification of the proximal phalanx of posterior digit - 2 in the 500 mg/kg/day treatment group as compared to the control group. However, these decreases were not considered to be biologicall y s ignificant. Therefore, the developmental toxicity NOAE L is 500 mgfkglday based on no adverse develop menta l effects at the highest dose administered. The developmental toxicity L OAEL was >500 mg/kg/day . In a 28 - day

repeated dose dermal toxicity study (MRID 44014233) , groups of 5 fe males Tif: RAIf (SPF) albino rats were treated with acibenzolar - S - methyl technical in 0.5% (w/v) carbox ym eth ylcellulose/0.1% (w/v) aqueous polysobate 80 at do ses of 0, 1, 10, 100 or 1000 mg/kg/day . Animals were treated b y dermal occlusion for 6 hours/day , 5 day s/week for 4 weeks. No animals died during the study . There were no treatment - related clinical signs, dermal effects, effects on body weight, food consumption, hematology , clinical chemistry , or organ weight. No gross or microscopic pathology were noted at necrops y. The sy stemic and dermal NOAEL is the lim it dose of 1000 mg/kg/day . A dermal and s ys temic LOAELwere not identified. **870.3800 Reproduction** In a 1 - generation range - finding reproduction study in rats (Nonguideline study , MRID 44014241 ) CGA 245704 Technical (97.9% a.i.; Batch No. P.303011) was administered to groups of 15 male and 15 female Tif: RAT f (SPF), h ybrids of RTTI1 x P11/2 (Spra gu e - Dawley derived) rats at dietary

concentrations of 0, 2000, 4000, or 8000 ppm for one generation (MR I D 44681301). In this range - finding study, F0 adults we re administered test or control diet for two weeks prior to mating. Premating doses were 199.4, 382.0, and 700.1 mg/kg/ day , respectively , for F0 males and 208.7, 409.9, and 727.8 mg/kg/day , respectivel y, for F0 females. Males were terminated at the end of mating and females were terminated at weaning of their pups. All offspring, except five litters per dose group, were necropsied at weaning. After weaning, five litters per dose group were retained on treatment for an additional three weeks and then necrops ied. The only

clinical sign of toxicity

in the parental animals was perineal hemorrhage during late gestation in five high - dose females. Gross necrops y was unremarkable. Absolute body weights, body weight gains, and food consumption values for the low - an d mid - dose males and low - dose females were similar to the controls throughout the study . High - dose males had significantl y (p > 0.01; 88 - 91% of control) lower body weights than the controls beginning on day 8 and continuing until study termination. Overall weight gain by the high - dose males was 69% of the control level. Body weights and body weight gains of the high - dose females were significantl y (p > 0.01; body weights 88 - 90% of controls) Tess than the controls during the premating interval. Overall body weight gain by the high - dose group was only 30% of the control group level. Mid - dose females had significantly (p > 0.05) lower body

weight gains than the controls during day s 1 - 8. Reduced body weight gains in the high - dose males and females and mid - dose females correlated with significantly (p > 0.05 or 0.01) reduced food consumption by these treated groups during study day s 1 - 8. Lower body

weights, body weight gains, and food consumption continued for the mid - and high - dose females during gestation and lactation. BodyPage 42 of 47

weights of the high - dose group were significantl y (p > 0.05 or 0.01) lower than the controls throughout gestation with overall weight gain b y the high - dose females during gestation 27% (p > 0.01) of the control group level. Mid - dose females had significantl y (p > 0.05 or 0.01) lower body weights than the controls on GD 21 and on lactation day s 7, 14, and 21. Weight gains by the mid - dose group during gestation and lactation were 92% and 83%, respectivel y of the control group levels. Food cons umption by the high - dose group was significantl y (p > 0.05 or 0.01) less than the controls throughout gestation. Food consumption by the mid - dose group was significantl y (p > 0.01) less than the controls throughout lactation. Therefore, the LOAEL for parental toxicity

is 4000 ppm based on reduced food consumption and lower body weight gains in F0 females. The parental toxicity NOAEL is 2000 ppm. Mating and fertility indices, mean precoital interval, mean gestation length, and mean numbers of corpora lutea and implantation sites per dam were similar between the treated and control groups. Mating and fertility indices for the high - dose group were similar to the control group, but no live lifters were produced by the high - dose group (i.e. all pregnancies were lost). L ive births and pup survival were similar between the low - and mid - dose groups and the control group. Therefore, the LOAEL for reproductive toxicity

is 8000 ppm based on total resorptions in all dams. The reproductive toxicity NOAEL is 4000 ppm. No treatment - related clinical signs of toxicity were observed in the pups during lactation. Absolute body weights of pups in the low - dose group were similar to the controls throughout lactation. Body weights of the mid - dose pups were significantl y (p > 0.05 or 0.01; 76 - 89% of controls) less than the controls beginning on lactation day 7 and continuing throughout lactation and postweaning. The lower body weights of the mid - dose pups were a result of significantly (p > 0.05 or 0.01; 69 - 89% of controls) lower bo dy weight gains from postnatal day

4 through postnatal day

28. Recovery

of body weight gains occurred during the postweaning intervals of day s 28 - 35 and 35 - 42. Therefore, the LOAEL for developmental toxicity is 4000 ppm based on reduced pup body weight gai ns and lower pup body weights during lactation and early postweaning. The developmental toxicity NOAELis 2000 ppm. **870.6200b Subchronic Neurotoxicity** In a subchronic neurotoxicity study (MRID 45713601 ), CGA - 245704 ( Acibenzolar, 97.9% a.i., Batch No. P.303011 ) was administered in the diet to 10 Sprague - Dawley derived rats/sex/gr oup at dose levels of 0, 400, 2000, or 8000 ppm (equivalent to 0/0, 24/26, 126/143, and 575/628 mg/kg/day

[M/F], respectively) for at least 90 days. Neurobehavioral assessment (functional observational battery

[FOB] and motor activity testing) was performed using all rats at pre dosing and Weeks 4, 8, and 13. At study termination, 5 rats /sex/group were anesthetized and perfused in situ for neuropathological examination. The t issues from the perfused animals in the control and 8000 ppm groups were subjected to histopathological evaluation of brain and peripheral nervous s ys tem tissues. Positive control data were not provided. No compound - related effects were observed in mortali ty , clinical signs of toxicity , FOB, motor activity , or gross or neuropathology . At 8000 ppm, body weights were generally

decreased throughout the study in the males (decreased 11 - 14%, not statistically significant) and females (decreased 8 - 13%, attaining statistical significance at Weeks 2 - 5, 7 and 8, and 11 - 13). Also at this dose, cumulative body weight gains were decreased throughout the study in the males (decreased 19 - 35%, attaining statistical significance at Weeks 2 - 4 and 6 - 9), and in the females ( decreased 24 - 40%).Page 43 of 47

A decreasing trend (p<0.01) in body weight and body

weight gain was also generall y noted in both sexes at this dose. These decreases in body weight gain corresponded with decreases in food consumption which was sporadically decreased i n the 8000 ppm males (decreased 9 - 21% at Weeks 1 - 3, 5 - 7, and 11, but only attaining statistical significance during Week 1), and females (decreased 14 - 18%, p<0.01 at Weeks 1 and 3). A decreasing trend (p<0.01) in food consumption was also generall y noted in the males at 8000 ppm during the weeks reported above. No neurological effects were observed at any dose in either sex. The LOAEL was 8000 ppm (equivalent to 575/628 mg/kg/day , M/F) based on decreased body weights, body weight gains, and food consumpti on . The NOAEL is 2000 ppm (equivalent to 126/143 mg/kg/day , M/F) . The subchronic neurotoxicity study in rats (MRID 45713601) on Acibenzolar had been reviewed and is classified as acceptable/non - guideline. **870.5100 ,**  **870 -**  **5300 , 870.5375, 870.5900**  **Mutagenicity** Five acceptable mutagenicity studies on technical grade acibenzolar - S - methyl are available: 1. Reverse gene mutation, S. ty phimuriumlE. Coli (MRID 44014247) 2. Forward gene mutation, Chinese hamster V79 lung cells in culture/HGPRT locus (M RI D 44014246) 3. Chromosome aberration, Chinese hamster ovary (CHO) cells in culture ( MRID 44014245) 4. Unscheduled DNA s ynthesis assay, primary rat hepatocy tes in culture (MRID 44014248) 5. In vivo cy

togenetics, micronucleus assay in mice (MRID 4401 4244) Results in all five studies were negative for genotoxicity . The five studies on the technical grade product satisf’ the new revised mutagenicity guideline requirements for a new chemical (published in 1991). Three additional acceptable mutagenicity studies on a) an iso mer (CGA - 362020) of acibenzolar S - methyl, b) a b y - product (NOA 419191) of acibenzolar - S - methyl, and c) a plain metabolite (CGA - 323060) of acibenzolar - S - methyl are also available. 1. Reverse gene mutation, S. ty phimuriumlE. Coli; Test Material: CGA - 362020 (MRID 44537025). This study

was POSI TIVE for evidence of induced mutant colonies over background in S. ty

phimurium strain TA1537 at concentrations of 277.8 and higher in the absence of 59 - mix. 2. Reverse gene mutation, S. ty phimuriumlE. Coli; Test Material: NOA 419191 (MRID 44537024). This study

was negative. 3. Reverse gene mutation, S. ty phimuriumfE. Coli; Test Material: CGA - 323060 (M RlD 44537026). This study

was negative CGA - 362020, an isomer of acibenzolar - S - methyl, is a low level impurity arising from a new production process for acibenzolar - S - methyl, which has not yet been submitted to the Agency .Page 44 of 47

Trace levels of this isomer (<0.1%) were found in some batches of technical grade acibenzolarS - niethyl produced via th e new production process. See the 6(a)(2) letter from Novartis, dated 3/16/98, reporting the findings in this mutagenicity study and the above information regarding the formation of this isomer in the new production process. Presumabl y, the same isomer doe s not occur in the old production process which produced the test material (lot/batch # P.303011) that was used for virtually all the toxicity testing on acibenzolar - S - methyl up to this point in time.Page 45 of 47

**APPENDIX B .**  **Chemical Names And Structures Of Metabolites** 

N N S S C H 3 O

Acibenzolar N N S C O OH

CGA 210007

N N S C O OH

HO CGA 324041 N N S C O OH

OH CGA 323060 S CO OH

O C H

3 CGA 379019

CGA 379789 S CO OH

O C H

3 O CGA 379018 S C O OH

C H

3 TRR, % ppm Lettuce 0. 6 0. 006 Tomato Fruit 7. 3 0. 023 Leaves 1. 5 0. 011 Wheat Forage 2. 4 0. 007 Straw 1. 3 0. 004 G rai n 4. 8 <0. 001 Ri ce Straw 0. 5 0. 009 TRR, % ppm Lettuce 20. 1 0. 204 Tomato F ru it 4. 8 0. 015 Leaves 6. 1 0. 044 Wheat Forage 2. 4 0. 007 S traw 1. 3 0. 004 G ra in 4. 8 <0. 001 TRR, % ppm Lettu ce 25.1 0.255 Tomato Fruit 65.2 0.203 Leaves 19.2 0.138 Wheat Forage 57.9 0.168 Straw 43.5 0.149 G rai n 39.3 0.001 Rice G rai n 5.6 0.005 Hul ls 16.8 0.027 Straw 56.2 1.120 TRR, % ppm Lettuce 16. 9 0. 171 Tomato F ru it 0. 8 0. 002 Leaves N D N D Wheat N D N D Ri ce N D N D

TRR, % ppm Rice G rai n 18.8 0. 016 Hul ls 7.9 0. 013 Straw 8.8 0. 175 al so found i n paddy water

TRR, % ppm Ri ce G ra in 3. 6 0. 003 S traw 0. 1 0. 002 also found i n paddy water TRR, % ppm Rice Straw 0.7 0. 014 al so found i n paddy waterPage 46 of 47Page 47 of 47

**APPENDIX C. Physical/Chemical Properties**  **Physicochemical Properties of Acibenzolar**  **- S**  **- Methyl**  **. Param**  **eter Value**  **Reference** Melting point/range 133 oC DP# 250963, 2/10/99, H. Podall pH 7 . 9 (1% solution in water) at 25 o C Density 1.54 g/cc at 22 o C (bulk density) Water solubility 0.0077 g/L Solvent solubility Methanol 4.2 g/L Acetone 28 g/L Toluene 36 g/L n - Octanol 5.4 g/L n - Hexane 1.3 g/L Ethyl acetate 25 g/L Dichloromethane 160 g/L European Commission, 6506/VI/99 - final, 5/21/02

Vapor pressure 3.6 x 10 - 6 Torr at 25 oC DP# 250963, 2/10/99, H. Podall Dissociation constant, pK a No dissociation constant in an accessible pH range (1.0 - 9.0) European Commission, 6506/VI/99 - final, 5/21/02 Octanol/water partition coefficient, Log(KOW ) Log ( P OW ) 3.1 at 25 oC DP# 250963, 2/10/99, H. Podall UV/visible absorption spectrum 253 nm ε : 14.7 10 3 l mol - 1 cm - 1 288 nm ε : 5.4 10 3 324 nm ε : 7.2 10 3 European Commission, 6506/VI/99 - final, 5/21/02 **APPENDIX D. Studies Reviewed for Ethical Conduct** The PHED Task Force, 1995. The Pesticide Handlers Exposure Database, Version 1.1. Task Force members Health Canada, U.S. Environmental Protection Agency , and the National Agricultural Chemicals Association, released February , 1995. Agricultural Re - entry

Task Force (ARTF) data ba se (SOP #3.1)