P age 1 of 53 UNITED STA

TES ENVIRONMENTA L PROTECTION AGENCY WASHINGTON, D.C. 20460 OFFICE OF PREVENTION, PESTICIDE

AND TOXIC SUBSTANCES MEMORANDUM August 1 9 , 2010 SUBJECT: Acibenzolar - S - methyl Human Health Risk Assessment for Propos ed Use of

Acibe nzolar - S - methyl on Turf and Cotton S eed. PC Code: 061402 DP Barcode: 379425 Decision No.: 422406 Registration No. : 100 - RGAG Petition No.: NA Regulatory Action: Section 3 Registration Risk Assessment Type: Single Chemical Aggregate Case No.: 7031 T XR No.: NA CAS No.: 135158 - 54 - 2 MRID No.: NA 40 CFR: 180. 561 FROM: Becky

Daiss , Biologist James Scott Miller , Environmental Scientist Bonnie Crop p - Kohlligian , Environmental Scientist Risk Assessment Branch 4 Health Effects Division (7509 P ) THROU GH: Susan V. Hummel , Branch Senior Scientist

Risk Assessment Branch 4 , HED (7509 P ) TO: Janet Whitehurst, Risk Manager Review

Fungicide Branch Registration Division (750 5 P) This document provides the Health Effects Division’s (HED’s) risk assessment of use of acibenzolar - S - methyl on turf and cotton seed . S upporting document s are listed below. Occupational Exposure Assessment

– J. S. Miller , D245704 , 8 / 19 /2010 Residue Chemistry

Assessment

– B. Crop p - Kohlligian , D3 73132 , 4 /2 9 / 10 Dietary and Drinking W ater Exposure Assessment

– B. Daiss, D 380742, 8 / 19 / 10 Drinking Water Exposure Assessment

– R. Miller, D 380135 , 7/9 / 10P age 2 of 53

TABLE OF CONTENTS pg. 1.0 EXECUTIVE SUMMARY ................................ ................................ ................................ .. 4 2.0 IN GREDIENT PROFILE ................................ ................................ ................................ .... 7 2.1 Registered Products ................................ ................................ ................................ ............. 7 2.2 Registere d and Proposed New Uses ................................ ................................ ..................... 7 2.3 Structure, Nomenclature and Physical/Chemical Properties ................................ ............... 7 3.0 HAZARD CHARACTERIZATION/ASSESSMENT ................................ ........................ 9 3.1 Hazard Characterization ................................ ................................ ................................ ...... 9 3.2 FQPA Hazard Considerations ................................ ................................ ........................... 1 0 3.2.1 Developmental Toxicity ................................ ................................ ....................... 1 0 3.2.2 Reproductive Toxicity ................................ ................................ ........................... 1 2 3.2.3 Developmental Neurotoxicity ................................ ................................ ................ 1 3 3.2.4 Pre - and/or Postnatal Toxicity ................................ ................................ ................. 1 4 3.2.5 Immunotoxicity ................................ ................................ ................................ ...... 1 4 3.2.6 FQPA Factor ................................ ................................ ................................ .......... 1 4 3.3 Toxicity

Endpoint Selection ................................ ................................ .............................. 1 4 3.3.1 Acute Reference Dose

– General Population ................................ ........................ 1 4 3.3.2 Chronic Reference Dose

– Females Age 13 - 49 & Young Children ...................... 1 5 3.3.3 Chronic Reference Dos e – Adult Males and Females Age 50+ ............................. 1 6 3.3.4 In cidental Oral Exposure ................................ ................................ ....................... 1 7 3.3.5 Dermal Absorption ................................ ................................ ................................ . 1 8 3.3.6 Dermal Exposure ................................ ................................ ................................ ... 1 9 3.3.7 Inhalation Exposure ................................ ................................ ............................... 19 3.3.8 Classificat ion o f Carcinogenic Potential ................................ ................................ 18 3.4 Margins of Exposure ................................ ................................ ................................ .......... 19 3.5 Recommendation for Aggregate Exposure ................................ ................................ ........ 19 3.6 Summary

of Endpoints Selected for Risk Assessment ................................ ...................... 20 3.7 Endocrine Disruption ................................ ................................ ................................ ......... 21 4 .0 DIETARY AND DRI NKI NG WATER EXPOSURE ASSESSMENT ............................ 2 2 4 .1 Residue Chemistry Profile ................................ ................................ ................................ . 2 2 4 .1.1 Metabolism in Primary Crops ................................ ................................ ................ 2 2 4 .1.2 Residue Analy

tical Method ................................ ................................ .................... 2 2 4 .1.3 Residues in Crops ................................ ................................ ................................ .. 2 3 4 .2 Drinking Water Profile ................................ ................................ ................................ ...... 2 3 4 .2.1 Environmental Fate and Transport ................................ ................................ ......... 2 4 4 .2.2 Estimated Drinking Water Concentrations ................................ ............................ 2 3 4 .3 Dietary

and Drinking Water Exposure and Risk ................................ ................................ 2 4 4 .3.1 Acute Dietary

and Drinking Water Analysis ................................ ......................... 2 5 4 .3.2 Chronic Dietary

and Drinking Water Analysis ................................ ...................... 2 5P age 3 of 53

5.0 RESIDENTIAL EXPOSURE AND RI SK ASSESSMENT ................................ ............. 26 5.1 Exposure Scenarios ................................ ................................ ................................ ........... 26 5.1.1 Handler Exposure Scenarios ................................ ................................ ................... 26 5.1.2 Post - Application Exposure Scenarios ................................ ................................ ..... 26 5.2 Exposure Data and Assumptions ................................ ................................ ...................... 26 5.2.1 Handler Exposure Scenarios ................................ ................................ ................. 27 5.2.2 Post - Application Exposure Scenarios ................................ ................................ .. 27 5.3 Residential Exposure and Risk Estimates ................................ ................................ ......... 28 5.3.1 Handl er Exposure and Risk ................................ ................................ .................. 28 5.3.2 Post - Ap plication Exposure and Risk ................................ ................................ .... 28 6 .0 AGGREGATE EXPOSURE AND RI SK ASSESSMENT ................................ ............... 29 6.1 Acute and Chronic Aggregate Risk ................................ ................................ ................... 30 6.2 Short Term Aggregate Risk ................................ ................................ .............................. 30 7 .0 OCCUPATI ONAL EXPOSURE AND RI SK ................................ ................................ ... 30 7 .1 Exposure Scenari os ................................ ................................ ................................ ............ 30 7 .1.1 Handler Exposure Scenarios ................................ ................................ .................. 31 7 .1.2 Post Application Exposure Scenarios ................................ ................................ .... 31 7 .2 Occupational Exposure Data and Assumptions ................................ ................................ . 31 7 .2.1 Exposure Data ................................ ................................ ................................ ........ 31 7 .2.2 Exposure Assumptions ................................ ................................ .......................... 3 2 7 .3 Occupational Exposure and Risk Estimates ................................ ................................ ..... 3 3 7 .3.1 Han dler Exposure and Risk Estimates ................................ ................................ ... 3 3 7 .3.2 Post - Application Exposure and Risk Estimates ................................ ..................... 3 4 8 .0 CUMULATIVE RI SK ................................ ................................ ................................ ....... 3 5 9 .0 DATA NEEDS ................................ ................................ ................................ ................... 3 6 9 .1 Toxicity

Data Requirements ................................ ................................ .............................. 3 6 APPENDI CES 1.0 GUIDELINE TOXICOLOGY DATA SUMMARY ................................ .......................... 3 7 1.1 Guideline Data Requirements ................................ ................................ ................ 3 7 1.2 Toxicity

Profiles ................................ ................................ ................................ ..... 3 8 2.0 STUDIES REVIEWED FOR ETHI CALCONDUCT ................................ ...................... 43 3 .0 ADDI TI ONAL TOXICITY STUDIES ................................ ................................ .............. 44 4 .0 REFERENCES FOR TOXI CI TY STUDIES ................................ ................................ ..... 50 5.0 CHEMI CALNAMES AND STRUCTURES OF METABOL IT ES ................................ . 53P age 4 of 53

1.0 EXECUTIVE SUMMARY This assessment provides inf ormation to support an amend ed Section 3 registration for the use of acibenzolar - S - methyl on cucurbit cotton and turf . This document addresses the exposures and risks associated with exposures from currently registered uses and the proposed new use s of ac ibenzolar - S - methyl. 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 leafy vegeta bles, Brassica leafy

vegetables, fruiting vegetables, 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) . Section 18 t ime - limited tolerances are established under §180.561(b) for bulb and green onion (0.05 ppm) . Acibenzolar - S - methyl acts to protect plant s against invading pathogen by induc ing a natural defense reaction that enhances the plants ability to ward off disease. Applications may be made using ground or aerial equipment. The proposed new use on turf grass is the first use of acibenzolar - S - methyl products intended for homeowner use and application to residential areas. Proposed New Use s Sy ngenta Crop Protection, Inc. has submitted an application for the registration of two new product s, Bion TM 500 FS and Daconil ACTion TM

. Bion TM 500 FS ( EPA Reg. No. 100 - RGAU ) i s a flowable concentrate formulation containing 42.0% acibenzolar - S - me thyl (equivalent to 4.17 lb ai/gal). It is proposed for use in commercial seed treatment operations for protection against certain early season seed and seed

–

borne nematodes in cotton . Daconil ACTion TM ( EPA Reg. No. 100 - RGAG ) is also a f lowable formul ations co ntaining 11% acibenzolar - S - methyl) . Itis proposed for use on sod farm s , golf course s , collegiate athletic fields, and lawns around commercial and industrial buildings to control turf diseases . The Daconil ACTion TM product also contains 54% chlo rthanalonil, however, only acibenzolar - S methyl exposures are evaluated for this assessment. Hazard Identification T he toxicology

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

and selecting endpoints for pur poses of this risk assessment. Acibenzolar - S - methyl showed no significant toxicity in a battery

of acute toxicity

tests (Toxicity Category I II or IV in all tests) but showed considerable skin - sensitivity . The end - use product (Actigard TM 50Wg) displaye d no significant acute toxicity and no dermal sensitizing potential. Insubchronic 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 obse rved in these studies included decreases in body weight, body

weight gain and/or food consumption. No other significant treatment - related effects of toxicological concern wereP age 5 of 53

observed in these subchronic and chronic oral studies. No neurotoxic effects w ere seen at the highest dose tested in a subchronic neurotoxicity study in rats. Ina 28 day

dermal toxicity

study in rats no sy stemic or dermal effects were seen at the limit dose. Ina 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 developmental 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 - methylwas classified by HED’s Hazard I dentification Assessment Review Committee ( HIARC) as a “not likely” human carcinogen based on negative carcinogenicity studies in male and female rats and mice and negative results in an acceptable battery of genotoxicity studies. An immunotoxicity

study is required as part of new 40 CFR Part 158 the data requirements for registration of a pesticide. A b attery of mutagenicity

studies performed on technical - grade acibenzolar - S - methyl prepared by the thi azole production process and an Ames assay using the technical - grade acibenzolar - S - methyl prepared by the thi azole production process are required . Dose Response Assessment Toxicological endpoints were selected for dietary /drinking water , occupational and residential exp o sure scenarios . Acute and chronic reference doses (RfDs) were selected for assessment of food and drinking water exposures. An acute RfD for the general population was not selected because no effect attributable to a single day oral exposure was observe d in animal studies. A cute and chronic RfD ’s for females 13 - 49 w ere selected from a developmental neurotoxicity toxicity

study in rats . A chronic RfD for the general population was selected from a c hronic feeding study

in dogs with chronic studies in oth er species co - critical . Short and intermediate - term occupational exposures via the dermal and inhalation route s may occur based on the use pattern and label directions . T oxicological endpoint s for inhalation , dermal , and incidental oral exposure s w ere se lected from the developmental neurotoxicity 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) , occupational , and residential exposure pathway s based on registered uses and request s for new use s of acibenzolar S - methyl on cucurbit vegetable and bulb onions . Although both of these products are designated for occupational/commercial use only , labels stipulate use on golf course s and on lawns around commercial buildings which may include apartment complexes . T herefore residential golfer and post application exposure scenarios were assessed. W orker s exposures were assessed for handlerP age 6 of 53

and post - application activities. Residential exposures were assessed for adult handlers and adult and child post - application activities. Refined acute and screening level chronic dietary and drinking water risk assessments for acibenzolar - S - methyl conclude that dietary and drinking water exposure estimates are below HED’s level of concern the general population and all population subgroups . Occupational and residential e xposure and risk esti mates indicate that worker and residential handler and post - application exposures are not of concern at the maximum allowable application rate s for the proposed new use s. Use of Human Studies This risk assessment relies in part on data from studies in which adult human subjects were intentionally

exposed to a pesticide or other chemical. These studies , listed in Appendix 2.0 , have been determined to require a review of their ethical conduct . S ome of these studies are also subject to review by

the Hum an Studies Review Board. All of the studies used have received the appropriate review. Environmental Justice Potential areas of environmental justice concerns, to the extent possible, were considered in this human health risk assessment, in accordance w ith U.S. Executive Order 12898, "Federal Actions to Address Environmental Justice in Minority Populations and L ow - In come Populations," http://www.eh.doe.gov/oepa/guidance/justice/eo128 98.pdf ). As a part of every

pesticide risk assessment, OPP considers a large variety of consumer subgroups according to well - established procedures. Inline with OPP policy , HED estimates risks to population subgroups from pesticide exposures that are b ased on patterns of that subgroup’s food and water consumption, and activities in and around the home that involve pesticide use in a residential setting. 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 considerations are currently in development, as OPP has committed resources and expertise to the development of spe cialized software and models that consider exposure to bystanders and farm workers as well as lifestyle and traditional dietary patterns among specific subgroups. Food Use Recommendation Based on the results of the metabolism (radiotracer) study , condu cted at the maximum proposed use rate, which demonstrated that the total transferable residue ( TRR) levels in/on all raw agricultural commodities of cotton were < 5 ppb, HED concludes that the proposed use of BionTM 500FS as a cottonseed treatment meets EPA ’s criteria for nonfood/nonfeed use classification, as defined in EPA’s Chemistry Science Advisory

Council’s (ChemSAC’s) 28 October 1999 memorandum, Classification of Seed Treatments as Food or Nonfood Uses. No additional data on residues of acibenzolar - S - methyl in/on cotton grown from treated seed are needed to support the proposed use.P age 7 of 53

2.0 INGREDIENT PROFILE 2.1 Registered Products There are currently 7 active acibenzolar - S - methyl registrations including two Section 3 registrations and five speci al local needs (SL Ns) or 24(c) registration s. Table 1. Sum ma ry 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 5 0 FL030003 Actigard 50 WG Plant Activator 50 GA020005 Actigard 50 WG Plant Activator 50 NC020007 Actigard 50 WG Plant Activator 50 SC030004 Actigard 50 WG Plant Activator 50 VA030002 Actigard 50 WG Plant Activator 50 2.2 Registered and Propose d New Uses Acibenzolar - S - methyl is a “plant activator” that protects the plant against invading pathogens by

stimulating the plant’s natural defense mechanisms. The single registered formulation, Actigard® (EPA Reg. No. 100 - 922), is a water - dispersible granule (WG) containing 50% acibenzolar - S - methyl. Sy ngenta is requesting registration of two new products . Bion TM 500 FS ( EPA Reg. No. 100 - RGAU ) and Daconil ACTion TM ( EPA Reg. No. 100 - RGAG ). Bion TM 500 FS i s a flowable concentrate formulation co ntaining 42.0% acibenzolar - S - methyl (equivalent to 4.17 lb ai/gal). It is proposed for use in commercial seed treatment operations for protection against certain early season seed and seed - borne nematodes in cotton . Application rates range from 0.0003 mg ai to 0.0009 mg a.i per seed based on 5,000 seeds per pound. Daconil ACTion TM is a f lowable formulations co ntaining 11% acibenzolar - S - methyl (and 54% chlorthalonil ) proposed for use on sod farm s , golf course s , collegiate athletic fields, and lawns aroun d commercial and industrial buildings to control turf diseases. Application rates range from 4.0 to 5.4 fl oz of product per 1,000 sq. feet with a maximum single application rate of 0.022 lbs ai/Acre. Table 2 . Maxim um Application Rates for Acibenzolar - S - me thyl Existing and Proposed Uses

Application Site Max % AI Maxim um AR Reg No. L eafy vegetables, Brassica leafy vegetables, F ruiting V egetables, T obacco and Bulb Onion subgroup 3 - 07A 50 0. 03 lb/ai A 100 - 922 Cucurbit vegetable crop group 9 0.06 lb/ai A Turf Use 11 0.022 lb/ai A 100 - RGAG Cotton Seed 24 0.0009 mg/ai SeedP age 8 of 53

2. 3 Structure, Nomenclature, and Physical/Chemical Properties The nomenclature and physicochemical properties of acibenzolar - S - methyl are provided in Tables 3 and 4 . Tab le 3 . Acibenzolar - S - Methyl Nom enclature . Chemical structure

N N S O S C H 3

Common name acibenzolar - S - methyl Company experimental name CGA - 245704 IUPAC name benzo[1,2,3]thiadiazole - 7 - carbothioic acid - S - methyl 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 N N S C O O H

1,2,3 - benzothiadiazole - 7 - carboxylic acid Table 4 . 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(K OW ) 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, 5/21/02P age 9 of 53

3 .0 HAZARD CHARACTERIZATION/ASSESSMENT 3.1 Hazard Characterization 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 is required as part of new 40 CFR Part 158 data requirements for reg istration of a pesticide. An Ames assay and a battery

of other mutagenicity studies performed on technical - grade acibenzolar - S - methyl prepared by the thi azole production process are required. Acibenzolar - S - methyl showed no significant toxicity in a batt ery

of acute toxicity

tests (Toxicity Category I II 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 de monstrated no significant acute toxicity ; additionally , the end - use product did not show dermal sensitization in guinea pigs. Insubchronic and chronic oral studies in rats, dogs and mice, signs of mild regenerative hemoly tic anemia were consistently obs erved in all three species. These signs frequently included decreased ery

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

hematopoie sis in the spleen, and increased spleen weights in both males and females. A compensatory response (increased ery

throcy 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 - related 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 o bserved at dose levels up to 1000 mg/kg/day , the limit dose. No neurological effects were observed at any dose in a subchronic neurotoxicity

study in rats . T reatment - related developmental malformations, anomalies and variations were observed in a devel opmental toxicity study in rats at or below the no observable adverse effect level (NOAEL) for maternal toxicity . At the highest dose tested in 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 system 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 pup s (as compared to adults) was not observed in a developmental toxicity study in rabbits or in 2 - generation and 1 - generation (range - finding) stud ies in rats. In a dermal developmental toxicity

study in rats, no maternal or developmental toxicity was observ ed at dose levels up to 500 mg/kg/day , the highest dose tested. Ina metabolism study

in rats, acibenzolar - S - methyl was rapidly and nearly

completelyP age 10 of 53

absorbed and excreted in less than 48 hours. The major route of excretion was urine (88 - 95%). The m ajor metabolite in urine was the carboxylic acid derivative of parent (79 - 92%). Ina battery

of mutagenicity

studies, results were negative in all studies except 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. This finding, however, can only be considered to be suggestive evidence of a possible aneuploidy effect since negative results were observed in an in vivo mouse micronucleus ass ay . In a S. typhimurium reverse gene mutation assay

performed on a structural isomer (CGA - 362020) of acibenzolar - S - methyl, positive results were observed in strain TA1537 in the absence of S - 9 activation. T his isomer is found at low levels ( < 0.1%) in s ome batches of technical - grade acibenzolar - S - methyl manufactured by

a new “thiazole process.” Therefore, an Ames assay and a battery

of mutagenicity

studies performed on technical - grade acibenzolar - S - methyl prepared by that process are required. Ina 2 - ye ar chronic toxicity /carcinogenicity

study in rats and an 18 - month carcinogenicity study in mice, acibenzolar - S - methyl was negative for carcinogenicity when administered at dose levels adequate for the testing of carcinogenic potential. The complete toxicity

profile for acibenzolar - S - methyl is provided in Appendix 1.2, Tables 2 and 3 . 3 .2 FQPA Hazard Considerations 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 developmental toxicity studies in the rat and rabbit, acceptable multi - and single generation reproduction stud ies in the rat , and an acceptable developmental neurotoxicity study in the rat are available. An acceptable subchronic neurotoxicity study in the rat is also available in the database. An acute neurotoxicity study

is not available but not required at this time due to the availability of the subchronic neurotoxicity

study in which n o neurological effects were observed at any dose . 3.2.1 Developmental Toxicity 3.2.1.1 Rat Developmental Study Study 1 Ina developmental toxicity study (MRID 44014236), 24 pregnant Tif : RAI f ( Sprague Dawley derived) rats per group were administered acibenzolar - S - methyl by gavage in 0.5% (W/W) aqueous sodium carboxy methyl - cellulose (vehicle) at daily doses of 0, 10, 50, 200, or 400 mg/kg/day

on gestation days (GD) 6 - 15, inclusive. Controls were treated only with 0.5% sodium carboxy methyl - cellulose. On GD 21, all dams were sacrificed and subjected to gross necropsy and fetuses were examined for external, visceral, and skeletal malformations and variations. All animals survived to scheduled sacrif ice.P age 11 of 53

Treatment - related hemorrhagic perineal discharge was observed in the 400 mg/kg/day group. No other treatment - related clinical signs of toxicity were observed in any

animal. Interpretations of changes in body weight, body

weight gain and food consumption for the 400 mg/kg/day

animals were severely compromised because the data were based on only 6 animals (3 of which had 50% resorptions compared to the number of implantation sites). The maternal toxicity L OAEL is 400 mg/kg/day based on clinical signs of hemorrhagic perineal discharge. The maternal toxicity

NOAEL is 200 mg/kg/day . There were no differences between the control group and any treatment group for number of animals pregnant, mean number of corpora lutea per dam, or mean number of implant ation sites per dam. At 400 mg/kg/day (highest dose tested), treatment - related complete litter resorptions, increased postimplantation loss, decreased mean live fetuses/litter, decreased mean fetal body weights, and several developmental malformations, an omalies, and skeletal variations were observed. At 200 mg/kg/day , treatment - related developmental malformations and skeletal variations were also observed. The developmental toxicity L OAEL is 200 mg/kg/day based on developmental malformations, anomalies, and skeletal variations. The developmental toxicity NOAEL is 50 mg/kg/day . Study 2 Ina developmental toxicity study (MRID 45089701 ), acibenzolar - S - methyl in aqueous 0.5% carboxy

methylcellulose was administered via daily oral gavage in a dose volume of 10 mL/kg to 22 presumed pregnant Sprague - Dawley rats at doses of 0, 10, 75, 150, or 350 mg/kg/day

from gestation day s (GD) 6 - 15 . On GD 20, all dams were euthanized; each dam’s uterus was removed via cesarean section and its contents examined. Fetuses w ere examined for external, visceral, and skeletal malformations and variations. All dams survived until scheduled termination, and there were no clinical signs of toxicity during the study

or gross lesions at necropsy . Furthermore, there were no effects of treatment on maternal body

weights, body

weight gains, or food consumption. A maternal LOAEL was not observed. The maternal NOAEL is 350 mg/kg/day . There were no abortions, premature deliveries, or complete litter resorptions. There were no effects of treatment on the numbers of litters, live fetuses, early resorptions, or late resorptions, resulting in post - implantation loss values in the treated groups that were comparable to controls. There were no treatment - related effects on placental weights, fetal body weights, or skeletal ossification. The number of fetuses with skeletal variations was significantly increased (p<0.05) at 350 mg/kg/day (60) compared to controls (18). This increase was accounted for by increased (p<0.05) incidences of fetuses with rudimentary lumbar ribs (31 fetuses compared to 9 controls) and long lumbar ribs (24 fetuses vs 0 controls). There were no other treatment - related external, visceral, or skeletal variations. There were no treatment - related external, visceral, or ske letal malformations. The developmental L OAEL is 350 mg/kg/day based on increased incidences of rudimentary and long lumbar ribs. The developmental NOAEL is 150 mg/kg/day .P age 12 of 53

3.2. 1 .2 Rabbit Developmental Study Ina developmental toxicity study (MRID 440 14237), 17 pregnant Russian Chbb:HM rabbits per group were administered acibenzolar - S - methyl by gavage in an aqueous solution of carboxy methylcellulose (0.5% w/w) at doses of 0 (vehicle), 10, 50, 300, and 600 mg/kg/day on gestation day s (GD) 7 - 19, inclusiv e. On GD 29, all surviving does were sacrificed and all fetuses were weighed and examined for external malformations/variations and visceral abnormalities/ variations. Three 600 mg/kg/ day

does were found dead and three more were sacrificed moribund during GD 13 - 28. All exhibited moderate to extreme body weight loss and reduced food consumption, two had perineal bleeding, two had diarrhea, and one had reduced motor activity . At 300 mg/kg/day , one doe was found dead on GD 18. This animal showed body we ight loss and reduced food consumption prior to death. Gross necropsy findings in animals that died intercurrently

included stomach hemorrhage in three of the 600 mg/kg/day does and in the 300 mg/kg/day

doe. No mortality

or treatment - related clinical signs of toxicity were observed for the 10 and 50 mg/kg/day

treatment groups. There were no significant differences in maternal body weights, body

weight changes, and food consumption in the surviving animals from the treated and control groups at any time d uring gestation. Therefore, the maternal toxicity L OAEL is 300 mg/kg/day

based on mortality , clinical signs of toxicity , and substantially decreased body

weights and food consumption. The maternal toxicity NOAEL is 50 mg/kg/day . No treatment - related eff

ects were observed for gravid uterine weights, number of fetuses/li tter, pre - and postimplantation loss, numbers of corpora lutea/doe, number of implantations/doe, resorptions/doe, fetal body weights, or fetal sex ratios. At 600 mg/kg/day there was a slight but significant increase in asy mmetrically

shaped caudal vertebral centers (3 fetuses in 3 li tters compared t o 0 fetuses or affected li tters in the control group). No statistically significant differences in the incidence rates of any external or visce ral malformations/anomalies/ variations were observed in the treated litters as compared to controls. Therefore, the developmental toxicity

L OAEL is 600 mg/kg/day based on a marginal increase in vertebral anomalies. The developmental NOAEL is 300 mg/kg/d ay . 3 .2. 2 Reproductive Toxicity Ina 2 - generation reproduction toxicity study , acibenzolar - S - methyl was administered to groups of 30 male and 30 female Tif: RAT f(SPF), hy brids of RII/ 1 x RI I/2 (Sprague - Dawley derived) rats at dietary

concentrations of 0, 20, 200, 2000, or 4000 ppm for two generations (MRID 44014241). Two litters were produced in the first generation and one litter was produced in the second generation. F1 adults were chosen from the F 1a pups and weaned onto the same diet as their parents. Animals were given test or control diet for 10 weeks before mating within the same dose group. All animals were continuously exposed to test material either in the diet or during gestation and lactation unt il sacrifice. No treatment - related clinical signs of toxicity were observed in males or females of either generation at any

time during the study . L ower body weights and food consumption for high dose dams was observed during lactation. No dose - or tre atment - related effects on body weights,P age 13 of 53

body weight gains, or food consumption were observed for the F1 males or females in the 20, 200, or 2000 ppm groups. Absolute body weights were significantly reduced in high - dose F1 males throughout the study and in high - dose F1 females through study day 57. No significant differences in body

weights and food consumption for the F0 and F1 dams were observed during gestation. Absolute spleen weights were significantly increased in both the 4000 and 2000 ppm F0 and F1 animals . Relative spleen weights were significantly increased in the 2000 and 4000 ppm F0 and F1 males and females. High - dose F0 males also had significantly increased absolute and relative kidney

weights. A dose - related increase in the severity of spl enic hemosiderosis was observed in F0 and F1 males and females. This finding was associated with an increase in the incidence of splenic congestion in the high - dose groups. T he LOAEL for parental toxicity is 2000 ppm based on increased spleen weights and hemosiderosis of the spleen in the F0 and F1 males and females. The parental toxicity NOAEL is 200 ppm. Mating, fertility , gestation, and parturition indices, mean precoital interval, and mean gestation length were similar between the treated and control groups of both generations. Mean litter size, live births, and pup survival were also similar between the treated and control groups for both generations. Therefore, the NOAEL for reproductive toxicity is 4000 ppm (highest dose tested). A LOAEL for repr oductive toxicity was not observed (>4000 ppm). No treatment - related clinical signs of toxicity were observed in the F 1 a, F 1 b, or F2 pups during lactation. Body

weights of the 4000 ppm group F 1 a , F 1 b, and F2 pups were significantly less than the control s on lactation day s 4, 7, 14, and 21. Body weights of the F1a, F 1 b, and F2 pups in the 2000 ppm group were significantly less than their respective controls on lactation day 21. Body weight gains by

the high - dose F 1 a and F2 pups throughout lactation and by the high dose F 1 b pups beginning on lactation day 4 were significantly

less than the controls. The LOAEL for developmental toxicity is 2000 ppm based on reduced pup body weight gains and lower pup body

weights during lactation. The developmental toxic ity NOAEL is 200 ppm. 3.2. 3 Developmental Neurotoxicity Study Ina developmental neurotoxicity study (MRID 46046401) a cibenzolar - S - m ethyl w as administered in the diet to pregnant Wistar rats (30/dose) from gestation day (GD) 7 to lactation day (LD) 22 at nominal doses of 0, 100, 1000, or 4000 ppm (equivalent to 0/0, 8.2/15.5, 82.0/153.6, and 326.2/607.8 mg/kg/day [gestation/lactation]). Dams were allowed to deliver naturally and were killed on LD 29. On postnatal day (PND) 5, litters were standardized to 8 pups/litter; the remaining offspring and dams were sacrificed and discarded without further examinations. Subsequently , 1 pup/litter/gro up was allocated to subsets for FOB, motor activity , acoustic startle response, learning and memory evaluation, and neuropathological examination. A maternal LOAEL was not observed. The maternal NOAEL is 4000 ppm (326.2 mg/kg/day ). Inthe offspring, no treatment - related effects were seen on survival, clinical signs, FOB, developmental land marks, brain weights or neuropathology . No conclusions can be drawn on the effects of the compound on motor activity due the lack of consistency in the observations. At the high dose, treatment - related effects were decreased body weights and increased auditory startle response . No differences from controls were observed in any morphometric parameter in the treated animals on PND 12, except for increased length from m idline in the hippocampus ofP age 14 of 53

the 4000 ppm females. The following significant differences in various morphometric measurements were noted on PND 63: (i) decreased dorsal cortex 1 thickness at  100 ppm in males ; (ii) decreased dorsal cortex 2 thickness at  100 ppm in males; (iii) decreased thickness of the molecular layer of the cerebellum at the preculminat e fissure in the 4000 ppm males; (iv) decreased thickness of the molecular layer of the cerebellum at the prepy ramidal fissure in the  1000 ppm males; an d (v) increased thickness of the corpus callosum in the 4000 ppm females. The biological significance of the reduction in the dorsal cortex, which is not dose - dependent, is unclear. The offspring LOAEL is 100 ppm (82 mg/kg/day ) based on changes in brain morphometrics in the cerebellum. The offspring NOAEL is (8.2 mg/kg/day ). 3 .2. 4 Pre - and/or Postnatal Toxicity The toxicology

database for acibenzolar - S - methyl is sufficient for FQPA assessment. In the rat developmental toxicity study treatment related visceral malformations and skeletal variations were observed in fetuses at 200 mg/kg/day , the NOAEL for maternal toxicity . Inthe developmental neurotoxicity study , offspring toxicity was observed at 82 mg/k g/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 rabbit 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 acibenz olar - S - methyl. There are no residual uncertainties with regard to pre and/or postnatal toxicity . 3.2.5 Immunotoxicity There are no indications in the available studies that organs associated with immune function , such as the thy

mus and spleen, are affected by acibenzolar - S - methyl . While effects on the spleen were observed in association with hematologic effects, these were considered to secondary to the primary

effects on blood hematology . Effects on the thy mus were seen in only one study in one animal at a high dose (400 mg/kg/day ). An immunotoxicity

study is required. This is a new data requirement under 40 CFR Part 158 as a part of the data requirements for registration of a pesticide (food and non - food uses). 3.2. 6 FQPA Factor The HED Tox icology Science Advisory Committee (Tox SAC ) recommended that the 10X FQPA safety

factor be reduced to 1X since endpoints protective of sensitive populations are being used in the risk assessment. 3.3 Toxicity Endpoint Selection 3.3.1 Acute Reference Dose (aRf D) - General Population Selected Study : Developmental Neurotoxicity in R ats (MRID 46046401 ).P age 15 of 53

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 brain morphometrics in the cerebellum. Uncertainty

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 femal es 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 analy sis has been expanded to include children for this assessment.]

3.3. 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. Uncertainty

Factor (UF ): 1 00

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

/Endpoint : 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 offspring. [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 re garding the potential for post - natal toxicity , the chronic dietary analy sis the chronic dietary exposure analy sis has been revised to include an assessment of dietary risks to children using the NOAELfrom the DNT study . ] Acute RfD = 8.2 mg/kg (NOAEL ) = 0. 0.082 mg/kg/day (General Population) 1 00 (UF)P age 16 of 53

3 . 3 . 3 Chronic Reference Do se (RfD) – Adult Males and Females 50+ years Selected Stud ies : Co - critical studies Chronic Feeding Study in Dogs (MRID 44014234 ); Onco genicity

Study Mouse (MRID 44014235) ; Chronic/Onco genicity Study Rat (MRID 44014243) ; Reproduct ive Study Rat ( 44014241 ) Chronic Dog Study - Ina chronic oral toxicity study acibenzolar - S - methyl was administered to 4 beagle dogs per sex per dose level in gelatin capsules at levels of 0, 5, 25, or 200 mg/kg/day

for 52 weeks. No biologically significant toxic effects were obs erved 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 hematologica l effects, hemosiderosis of the liver and spleen, extraniedullary

hematopoiesis in the spleen, and increases in liver weight. Significant treatment related changes were found in clinical chemistry in the dogs fed the test, substance at 200 mg/kg/day , ( inc rease in triglycerides 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 albumin in both sexes at week 13. These clinical chemistry changes are suggestive of possible nutrit ional 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 . Oncogenicity

Mouse Study - Inan oncogenicity study acibenzolar - S - methyl w as administered in th e 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 rese rved for measurements of hematology parameters after 12 and 18 months of treatment. Treatment did not result in a significant decrease in survival. The overall food efficiency was decreased in females by

22% at 6000 ppm . The red blood cell count, hemogl obin, 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 weeks . 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 especially in females at 2000 and 6000 ppm. Macroscopic and microscopic findings consistent with a mild hemoly

tic anemia in both sexes inclu ded increased incidences of enlarged spleen , increased incidences of hemosiderosis in liver , spleen , and bone marrow . Incidence 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 mild hemoly

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

males; 10.8 mg/kg/day

females). Chronic RfD = (NOAEL) 8.2 mg/kg = 0.082 ( F emales 13 - 49 /Young Children ) (UF) 100P age 17 of 53

Chronic/Oncogenicity

Rat Study - Ina chronic toxicity /oncogenicity

study , acibenzolar S - methyl was administered in the diet to groups of 80 male and 80 female (Tif:RAI f (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 w ere 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 effec ts were observed at 20 , 200 or 2500 ppm for any parameter. Treatment related effects on b ody weight, weight gain, food consumption, and food efficiency were observed in both sexes at 7500 ppm . Other treatment - related effects at 7500 ppm included a slight hemoly tic anemia in both sexes as evidenced by

decreases in ery

throcy te count, increased mean corpuscular volume, elevated reticulocy te counts, increased serum bilirubin, increased severity 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 by a statistically

significant increase in spleen 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 food 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 Establishing cRfD : 25 mg/kg/day based on a NOAEL of 25 mg/kg/day

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

tic anemia with compensatory response. Uncertainty

Factor(s) : 100

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

/Endpoint/Uncertainty

Factor : T he 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 L OAEL 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 pically , a combination of NOAELs/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 observed across species at a relatively

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 assessme nt on the NOAEL from the dog chronic toxicity study using the L OAEL from the reproduction study

in rats for the basis of the endpoint (hemoly tic anemia). Chronic RfD = 25 mg/kg/day (NOAEL ) = 0. 25 mg/kg/day (Ad ult Male ) 1 00 (UF)P age 18 of 53

3 . 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. Uncertainty

Fa ctor (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. 3 . 3 . 5 Dermal Absorption There are no derma l absorption studies available with acibenzolar - S - methyl . However, the dermal absorption rate for acibenzolar - S - methyl is estimated to be 40 % b y comparing maternal toxicity

L OAEL ( 400 mg/kg/day ) from an oral developmental toxicity study in rats (MRID 440 14236) to the L OAEL (> 1000 mg/kg/day ) from a dermal toxicity

study in rats (MRID 440142 33 ). [Note: A dermal absorption factor of 10% was used in the previous risk assessment of acibenzolar - S - methyl (D361893, 3/3/09). The DAF has been revised to 40% bas ed on a reevaluation of the dermal and oral toxicity data.] 3 . 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 offsp ring 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. Uncertainty

Factor (UF ): 100

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

/En dpoint : 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 findi ngs from th e 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 10% should be used . This factor is based on the ratio of the dermal NOAEL of 1000 di vided by

the oral maternal L OAEL of 400 which equals 40 %.P age 19 of 53

3.3. 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 LOAEL of 82 mg/kg/day based on changes in brain morphometrics in the cerebellum. Uncertainty

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 used 3.3. 8 Classification of Carcinogenic Potential The Hazard I dent ification Assessment Review Committee (HIARC) classified acibenzolar - S - methyl as a “not likely” human carcinogen based on negative carcinogenicity studies in rats and mice and on generally negative mutagenicity

studies. [Note: The cancer classification has not been updated based on the EPA’s 2005 Guidelines for Carcinogen Risk Assessment . ] 3 . 4 Margins of Exposure A summary

of target Levels of Concern for risk assessment is provided in Table 5 . Table 5 . Target Levels of Concern/Margin of Exposure f or Acibenzolar - S - Methyl Route/Duration Short - Term (1 - 30 Days) Interm ediate - Term (1 - 6 Months) Long - Term (> 6 Months) Occupational (Worker) Exposure Dermal 100 100 N/A Inhalation 100 100 N/A Residential (Non - Dietary) Exposure Oral 100 N/A N/A Dermal 100 N/A N/A Inhalation 100 N/A N/A 3 . 5 Recommendation for Aggregate Exposure Risk Assessments 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 are 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 endpointsP age 20 of 53

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 expo sures to should be aggregated for acibenzolar - S - methyl , because the same oral toxicity endpoint was selected for both exposure routes . 3 . 6 Summary of Endpoints Selected for Risk Assessment Toxicological doses/endpoints selected for the acibenzolar - S - methyl risk assessment are provided in Tables 6 and 7 . Table 6 . Sum ma ry 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/FQPA

Safety Factors RfD, PAD, LOC for Risk Assessment Study and Toxicological Effects

Acute Dietary (General Population) NOAEL =

8.2 mg/kg/day UF A= 10x

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

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

Rat Developmental LOAEL = 82 mg/kg/day based on changes in brain morphometrics in the cerebellum in offspring. Maternal LOAEL = wa s not observed NOAEL = 4000 mg/kg/day HDT Chronic Dietary (Females 13 - 49 years & Young Children) NOAEL =

8.2 mg/kg/day UF A= 1 0x .

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

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

Rat Developmental LOAEL = 82 mg/kg/day based on changes in brain morphometrics in the cerebellum in offspring. Maternal LOAEL = was n ot observed NOAEL = 4000 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 & Mo use, 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 Study -

Rat Devel opmental LOAEL = 82 mg/kg/day based on changes in brain morphometrics in the cerebellum in offspring. Maternal LOAEL = was not observed NOAEL = 4000 mg/kg/day HDT Dermal Short Term (1 - 30 days) DAF = 10% NOAEL=

8.2 mg/kg/day UF A= 10x

UF H =10x Occupat ional

LOC for MOE = 100 Developmental Neurotoxicity Toxicity Study -

Rat Developmental LOAEL = 82 mg/kg/day based on changes in brain morphometrics in the cerebellum in offspring. Maternal LOAEL = was not observed NOAEL = 4000 mg/kg/day HDTP age 21 of 53 Table 6 . Sum ma ry 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/FQPA

Safety Factors RfD, PAD, LOC for Risk Assessment Study and Toxicological Effects

Inhalatio n Short Term (1 - 30 days) NOAEL=

8.2 mg/kg/day UF A= 10x

UF H =10x Occupational

LOC for MOE = 100 Developmental Neurotoxicity Toxicity Study -

Rat Developmental LOAEL = 82 mg/kg/day based on changes in brain morphometrics in the cerebellum in offspring. Maternal LOAEL = was not observed NOAEL = 4000 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 beginnin g of extrapolation to determine risk associated with lower environmentally relevant human exposures. NOAEL = no observed adverse effect level. LO AEL = lowest observed adverse effect level. UF = uncertainty factor. UF A = extrapolation from animal to hum an (interspecies). 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 abse nce of 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. Table 7 . Sum ma ry 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 Level of Concern

for Risk Assessment Study and Toxicological Effects Derma l Short (1 - 30 days) and Intermediate (1 - 6 mo nths) Term DAF = 4 0% NOAEL= 8.2 mg/kg/day UF A= 10x

UF H =10x Occupational LOC

for MOE = 100 Developmental Neurotoxicity Toxicity Study - Rat

Developmental LOAEL = 82 mg/kg/day based on changes in brain morphometr ics in the cerebellum in offspring. Maternal LOAEL = was not observed NOAEL = 4000 mg/kg/day HDT Inhalation Short (1 - 30 days) and Intermediate (1 - 6 mo nths) Term NOAEL= 8.2 mg/kg/day UF A= 10x

UF H =10x Occupational LOC

for MOE = 1 00 Developmental Neurot oxicity Toxicity Study - Rat

Developmental LOAEL = 82 mg/kg/day based on changes in brain morphometrics in the cerebellum in offspring. Maternal LOAEL = was not observed NOAEL = 4000 mg/kg/day HDT Cancer ( all routes ) A “not likely” human carcinogen

DAF = Dermal Absorption Factor 3 . 7 Endocrine Disruption EPA is required under the FFDCA, as amended by FQPA, to develop a screening program to determine whether certain substances (including all pesticide active and other ingredients) “may

have an effec t in humans that is similar to an effect produced by a naturally occurring estrogen, or other such endocrine effects as the Administrator may designate.” Following recommendations of its Endocrine Disruptor and Testing Advisory Committee (EDSTAC), EPA det ermined that there was a scientific basis for including, as part of the program, the androgen and thy roid hormone sy stems, in addition to the estrogen hormone sy stem. EPA also adopted EDSTAC’s recommendation that the Program include evaluations of potenti al effects in wildlife. For pesticide chemicals, EPA will use FIFRA and, to the extent thatP age 22 of 53

effects in wildlife may

help determine whether a substance may have an effect in humans, FFDCA authority

to require the wildlife evaluations. As the science devel ops and resources allow, screening of additional hormone systems may be added to the Endocrine Disruptor Screening Program (EDSP). 4 .0 DIETARY AND DRINKING WATER EXPOSURE ASSESSMENT 4 .1 Residue Chemistry Profile 4 .1. 1 Metabolism in Primary Crops The nature of the residues in plants has been adequately delineated based on metabolism studies with lettuce, tomato, rice, wheat, and tobacco. The HED MARC has determined that the residues of concern in plants for tolerance expression are residues conver tible to benzo(1,2,3)thiadiazole - 7 - carboxylic acid (CGA - 210007), expressed as acibenzolar - S - methyl. For purposes of risk assessment, residues of the 4 - and 5 - hydroxy metabolites (CGA - 324041 and CGA - 323060) of CGA - 210007 should also be considered. For thi s 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 - methyl does 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 8 provides a summary of the MARC decisions regarding residues of concern for acibenzolar - S - methyl . Table 8 : Summ ary of MARC D ecisions for Acibenzolar - S - me thyl Matrix Residues of Concern 1 For Risk Assessm ent 2 For Tolerance Expression Plants Residues of acibenzolar - S - methyl, benzo(1,2,3)

thia diazole - 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) R esidues of acibenzolar - S - methyl, b enz o(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 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 assessment 4 .1. 2 Residue Analytical Methods 4 .1. 2 .1 Enforcement Method s An adequate method, HPL C/UV Method AG - 671A, is available for tolerance enforcement. This method has undergone a successful tolerance method validation by the Analy tical Chemistry

Branch of BEAD. The method was forwarded to FDA for inclusion in PAM Volume I I. T he method consists of an initial hydroly sis with NaOH to convert acibenzolar - S - methyl to CGA - 210007 followed by methanol extraction. Residues are then diluted with HCl and purified by a series of solid - phase extraction steps. Prior to HPL C/UV analy sis, residues are partitioned into ethyl acetate, dried down, and redissolved in phosphoricP age 23 of 53

acid. This method has an L OQ of 0.02 ppm. The method includes optional detection via HPL C/MS, giving a means of residue confirmation. The HPL C/UV and L C/MS methods are adequate for collecting data on residues of acibenzolar - S - m ethyl in/on cucurbit vegetables and bulb onions, respectively . 4 .1 . 2 .2 Multiresidue Methods Data depicting the recovery of acibenzolar - S - methyl and its metabolite, CGA - 210007, using FDA Multiresidue Methods, were previously sub mitted. Acibenzolar - S - methyl yielded adequate response to GLC analysis and was completely recovered through Protocols D and E. Partial recovery

of CGA - 210007 was achieved via Protocol B after derivatization. 4 .1. 3 Residue s in Crops Based on the results of the metabolism (radiotracer) study , conducted at the maximum proposed use rate, which demonstrated that the TRR levels in/on all raw agricultural commodities of cotton were < 5 ppb, HED concludes that the proposed use of BionTM 500FS as a cottonseed tre atment meets EPA’s criteria for nonfood/nonfeed use classification, as defined in EPA’s Chemistry

Science Advisory

Council’s (ChemSAC’s) 28 October 1999 memorandum, Classification of Seed Treatments as Food or Nonfood Uses. No additional data on residues of acibenzolar - S - methyl in/on cotton grown from treated seed are needed to support the proposed use. 4 .2 Drinking Water Profile 4 .2.1 Environmental Fate and Transport Laboratory

and field studies indicate that a cibenzolar - S - methyl does not have a high potential to leach to ground water nor move to surface water based on its short resident times in soil and soil water. The major degradate observed was benzo[1,2,3]thiadiazole - 7 - carboxylic acid (CGA 210007) in the hydroly sis and soil photoly sis studi es (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 exhibiting greater potential than the parent for lateral or vertical movement aft er application. Acibenzolar - S - methyl mainly degrades by

photoly sis and microbial action. Aqueous and soil photolysis irradiated half lives for parent are 0.6 hours and 1 day , respectively . The aerobic soil metabolism , anaerobic aquatic metabolism and aer obic aquatic half lives are 5.3 hours , 4.0 and < 1 day(s) respectively . Field dissipation half - lives ranged from < 1 to 27 day s . Under aerobic soil, aerobic aquatic, and anaerobic 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 250C pH 3.6 - 3.8 v. 7.7 ppm at 250C pH 7.5 - 7.9). CGA 210007 has longer resident fi eld 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 considerably more mobile. Acibenzolar is not considered a bioconcentrate nor is it highly bioaccumulative.P age 24 of 53

4 .2.2 E stimated Drinking Water Concentrations Drinking water residues provided by the Environmental Fate and Effects Division (EFED ) ( D351609, R. Miller, 8/8/08; D380135, 7/9/10) were incorporated directly into the acute and chronic dietary

analy ses ( “water, d irect, all sources” and “water, indirect, all sources EFED generated the surface water and ground water estimates using the Pesticide Root Zone Model 3 (PRZM3; ver. 3.12.2; 2 - May- 2005)/Exposure Analy sis Modeling Sy stem (EXAMS; ver. 2. 98.04.06; 25 - Apr - 2005 ) and Screening Concentration in Ground Water (SCI GROW; Tier 1) models, respectively . The residues of concern for drinking water include acibenzolar - S - methyl and residues convertible to CGA - 210007. Based on the EFED drinking water an aly sis conducted for the proposed new use of acibenzolar - S - methyl on cotton and turf, EDWCs from these uses will be less than the EDWCs for the existing use on cucurbits. Therefore estimated EDWCs for use of acibenzolar - S - methyl on cucurbits were used for the drinking water assessment because this use 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 4.08 x 10 - 5 μg/L and CGA 210007 is not expected to exceed 0.557 μg/L . Surface water EDWCs for acibenzolar - S - methyl, which were used in the exposure assessment, are provided in Table 9 . CGA - 210007 drinking water residues were included in the dietary exposure assessment as aci benzolar - S - methyl equivalents. Table 9 . EDWCs for use in screening the Acibenzolar - S - Methyl H uman H ealth Risk A ssessment Drinking Water Source Chemical Acute (μg/L) NonCancer Chronic (μg/L) Cancer Chronic(μg/L) Surface Water Acibenzolar 1 0.74 0.1 0 0.05 Surface Water CGA 210007 2 14.21 9.48 5.47 1 Aerial application method 2 Degradation product formed after application 4 .3 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 pesticide. For acute and chronic dietary a ssessments, 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 population adjusted dose (PAD). The PAD is equi valent to the reference dose (RfD) divided by 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 hydroxy 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 ) .P age 25 of 53

4 .3.1 Acute Dietary and Drinking Water Analysis A refined (probabilistic) a cute dietary exposure analy sis was performed for the general population and all population subgroup s . The acute analy sis assumed a distribution of residues based on field trial data. Empirical and DEEM default processing factors were used to modify the f ield trial data. M aximum screening - level percent crop treated estimates were used for commodities for which data were available. I f no percent crop treated data were available, 100% crop treated was assumed. The acute analysis incorporated the 1 in 10 year peak surface drinking water estimate f

rom application of acibenzolar - S - methyl to cucurbit vegetables. 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 38% of the aPAD. The 99.9 th percentile aPAD for the general U.S. population was 18%. Table 1 0 : Summary of Acute Dietary Exposure and Risk for Acibenzolar - S - Methyl Population Subgroup aPAD (mg/kg/day) Acute (99 th Percentile) Exposure (mg/kg/day) % aPAD General U.S. Population

0.082 0.0145 18 All Infants (< 1 year old) 0.0136 17 Children 1 - 2 years old 0.0250 30 Children 3 - 5 years old 0.0307 3 7 Children 6 - 12 years old 0.0204 25 Youth 13 - 19 years old 0.0152 19 Adults 20 - 49 years old 0.0094 11 Adults 50+ years old 0.0112 14 Females 13 - 49 years old 0.0096 12 4 .3.2 Chronic Dietary and Drinking Water Analysis A conservative chronic dietary exposure analy sis 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 analy si s 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 protective chronic population adjusted dose (cPAD, 0.0 82 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%.P age 26 of 53

Table 1 1 : Summary of Chronic Dietary Exposure and Risk to Female and Child Population for Acibenzolar - S - Methyl Population Subgroup c PAD (mg/kg/day) C hronic Exposure (mg/kg/day) % cPAD Females 13 - 49 years old 0.082 0.0038 5 All Infants (< 1 year old) 0.0043 5 Children 1 - 2 years old 0.0093 11 Children 3 - 5 years old 0.0088 11 Children 6 - 12 years old 0.0059 7 Table 1 2 : Summary of Chronic Di etary Exposure and Risk to Adults 50+ and Adult Males (13+) Acibenzolar - S - Methyl Population Subgroup cPAD (mg/kg/day) Chronic Exposure (mg/kg/day) % cPAD General U.S. Population 0.25 0.004 5 2 Males 13 - 19 0.004 2 2 Males 20+ 0.0038 2 Adults 50+ years old 0.0040 2 5.0 RESIDENTIAL EXPOSURE AND RISK ASSESSMENT 5.1 Exposure Scenarios The proposed uses associated with this action are for application of acibenzolar - S - methyl to turfgrass on golf courses , collegiate athletic fields, and lawns around commercial and industrial buildings. Based on the use patterns specified on the label, acibenzolar - S - methyl could be used on lawns around apartment complexes . T herefore , standard residential lawn and turf handler and post - application exposure scenarios w ere assessed for the proposed new use . 5.1.1 Handler Exposure Scenarios Based on the number of seasonal applications indicated on this product label, and information provided by

the registrant, residential handler exposures are expected to be sho rt - (1 - 30 day s) and intermediate - term (1 - 6 months) in duration. T he following residential handler exposure scenarios were assessed : 1) Lo w Pressure Handwand (dry flowable and soluble liquid) formulation to Lawn/Turf. 2) Hose - end Sprayer (dry flowable and soluble liquid) formulation to L awn/Turf. 5.1.2 Post - Application Exposure Scenarios Dermal exposure was assessed for adult and child recreational golfer and residential activities on treated turf. Incidental oral exposure (hand to mouth, object to mo uth, and soil ingestion) was assessed for toddlers.P age 27 of 53

Based on the Agency's current practices, a quantitative post application inhalation exposure assessment was not performed for acibenzolar - S - methyl . However, volatilization of pesticides may

be a potent ial source of post application inhalation exposure to individuals nearby 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 Sc ientific Advisory Panel (SAP) in December 2009. The Agency received the SAP’s final report on March 2, 2010 and is in the process of evaluating the SAP report. The Agency may , as appropriate, develop policies and procedures to identify the need for and, subsequently , the way to incorporate post application inhalation exposure into the Agency's risk assessments. I f new policies or procedures are put into place, the Agency may revisit the need for a quantitative post application inhalation exposure assessm ent for acibenzolar - S - methyl . 5.2 Exposure Data and Assumptions Maximum application rates for all of the exposure scenarios assessed are based on information provided in the proposed acibenzolar - S - methyl label. Based on the label, the maximum singl e application rate is 0.22 lbs ai per acre . The post - application risk assessment is based on generic assumptions from the Recommended Revisions to the Residential SOPs, and recommended approaches by HED’ Science Advisory

Council for Exposure (ExpoSAC). Th e following standard exposure assumptions were used in estimating post - application risks . 5.2.1 Handler Exposure Scenarios

• Average body

weight of an adult handler is 6 0 kg.

• Exposure duration is short - term. • D ermal absorption factor = 40%

• Maximum application rates as determined by label

• SOP daily

volumes handled and/or area treated used for the scenarios assessed are 0.5 acres treated per day 5.2.2 Post - Application Exposure Scenarios

• Average adult body weight is 6 0 kg

• Average older chil d (golfer) body weight is 39 kg

• Average yo ung child body weight is 15 kg

• D ermal absorption factor = 40% Golfers & Homeowners : • Exposure is assumed to occur on the day of application (day

0)

• Turf Transferrable residue is 5% of the application rate for the fraction initially available.

• Transfer coefficient is 500 cm 2 /hour for golfers

• Exposure duration is 4 hours per day for golfers and 8 hours per day for homeowners Child Hand - to - mouth

• Exposure is assumed to occur on the day of app lication (day

0)

• S aliva extraction factor is 50 percent .P age 28 of 53

• S urface portion of hand put in mouth is 20 cm 2

. • H and - to - mouth exposure frequency is 20 times per hour . • E xposure duration is 2 hours for outdoor exposures . Toddler Object to Mouth Sce nario

• O bject to mouth transfer efficiency is equal to 20% of the application rate . • Surface area from mouthing turf or a small object is 25 cm 2

. Toddler Incidental Soil Ingestion Scenario

• S oil ingestion rate is 100 mg/day . • F raction of ai ava ilable in uppermost cm of soil (fraction/c m) is 100 percent based on soil incorporation into top 1 cm of soil after application . 5.3 Residential Exposure and Risk Estimates 5.3.1 Handler Exposure and Risk A target LOC or MOE of 100 is considered adequate residential exposure and risk. All residential exposures are assessed as short - term based on label prescribed uses. Exposure and risk estimates indicate MOEs are not of concern (MOEs > 100) at the maximum use rate for the residential post - application exposure s cenario assessed. A summary of residential handler exposure and risk calculations, assumptions, and results is provided in Table 13 . Table 13 : Re sidential Handler Exposure Assessment Acibenzolar - S - Methyl Application to Lawns/Turf Exposure Scenario 2 Appl ication Rate (lb ai/A) 3 Area

Treated (A/day) 1 Derm al

Unit Exposure ( m g/lb ai) Inhal Unit

Exposure (  g/lb ai) 5 Derm al

Daily Dose (m g/kg/d) 5 Inhal

Daily Dose (m g/kg/d) Short

Term MOE 4 Mixing, Loading Applying Low Pressure Handwand (Soluble Liquid) 0.022 0 .5 100 30 0.00 73 0.00000 6 1100 Mixing, Loading, and Applying Hose - end Sprayer (Dry Flowable) 11 17 0.000 8 0.000003 10000 1. Dermal unit exposur e values . Values are reported in the PHED Surrogate Exposure Guide dated August 1998. 2. Appl ication rates are based on maximum values found in propose d labels. D aily area treated is based on the area or gallons that can be reasonably applied in a single day for each exposure scenario of concern based on the application method and formulation/packaging type. (Standard EPA/OPP/HED values). 4. Daily Dose (mg/kg/day) Dermal = Unit Exposure * % Absp * Appl. rate * Area treated} / BW - 60 kg. {Where: dermal absorption. is assumed to be 40 %} . Inhalation = Unit Exposure * Area treated/BW 5. Short - Term MOE = NOAEL (8.2 mg/kg/day) /Dermal + Inhalation Daily Dose . The LOC is 100. 5.3.2 Post - Application Exposure and Risk A target LOC or MOE of 100 is considered adequate residential exposure and risk. All residential exposures are assessed as short - term based on label prescribed uses. Exposure and risk estimates indicate MOEs are not of concern (MOEs > 100) at the maximum use rate for the residential post - application exposure scenario assessed. A summary of residential post -P age 29 of 53

application exposure and risk calculations, assumptions, and results is provided in Table s 14 and 15 . Table 14 . Estim ated Acibenzolar - S - Methyl Exposure & MOEs for Post - application Derm al Exposure to Treated Lawn/ Golf Course Turf LOC/MOE = 100 Exposure Scenario AR (lb ai/A) TC (cm 2 /hr) Exposure Duration (hrs/day) D erm al d ose

(m g/kg/day) Derm al

MOE Adult Golfers 0.022 500 4 0.000 164 50000 Child Golfer 500 4 0.000 253 30000 Adult Lawn Activities 14500 8 0.00 135 6000 Child Lawn Activities 5200 8 0.00 194 4000 1 Application rate s are based on maximum values based on proposed label. 2 TTR or DFR (mg/cm 2 ) = Application Rate (lb ai/A) x CF (4.54E+5 mg/lb) x CF (2.47E - 8 A/ cm 2 ) x 5% (initial fraction of ai retained on turf) Application rates are based 3 TC cm 2 /hr = Transfer coeffic ients and associated activities (ExpoSAC Policy Memo #003.1) 4 Dermal Dose (mg/kg/day) = TTR (mg/cm 2 ) x TC ( cm 2 /hr) x 4 (hrs/day )/70 kg (body weight) 8 Dermal MOE = short - term endpoint for dermal (NOAEL 8.2 mkd)/Daily Dermal Dose Table 15. Post - applicati on Incidental Ingestion Risks to Toddlers Reentering Treated Law ns Exposure Scenario AR (lb ai/A) Daily Oral Dose (m g/kg/day) Oral MOE Aggregate Oral and Derm al MOE Hand - to - Mouth 0.022 0.00033 25000 9000 Object - to - Mouth 0.00008 100000 Soil Ingestion 0.000001 >1000000 Hand To Mouth Calculation PDR(t) (mg/day) = (HT E (t) (μg/cm 2 ) * SEF * SA * Freq * ED/1000 (μg/mg) where: PDR = Potential Dose Rate at time (t) attributable for activity in a previously treated area (mg/day) HTE = Hand Transfer Efficiency at time t = 5% of Application Rate ( μ g/cm2) Application rate = 0.2% ai, 3 seconds per cubic yard , 1.5

g/sec , all spray settles to turf (MRID 47119801 ) SEF = Saliva Extraction Factor (50%) SA = Surface Area of Two Fingers (20 c m 2 ) Freq = Frequency of Hand to Mouth Events (20) ED = Exposure Duration in hours (2 hr/day) t = Postapplication Day on which exposure is being assessed (day 0) MOE = Short Term Oral NOAEL (8.2 mkd) /Daily Oral Dose (mg/ kg/ day) Object to Mou th Calculation PDR(t) (mg/day) = (DFR(t) ( μ g/cm2) * SA/1000 ( μ g/mg) where: PDR = Potential Dose Rate at time (t) attributable for activity in a previously treated area (mg/day) DFR(t)= Dislogeable Foliar Residue at time t = 20% of Application Rate ( μ g/c m2) APR = 1.1 μ g/cm 2 SA = Surface Area of grass or toy mouthed by toddler (25 cm 2 day) t = Postapplication day on which exposure is being assessed (day 0) MOE = Short Term Oral NOAEL (8.2 mkd) / [Daily Oral Dose (mg/kg/ day) MOEs are reported to two significant figures Soil Ingestion Calculation PDR(t) (mg/day) = (SRt * I gR * CF1) where: PDR = Potential Dose Rate - nondietary ingestion rate from contact with treated surface (mg/day) SRt = Soil Residue on day "t" ( μ g/g) = Application Rate ( μ g/ cm2) * 1/cm * 0.67 cm3/g soil [1/cm is fraction of ai available in

uppermost cm of soil] IgR = Ingestion Rate of soil (mg/day); (100 mg/day) CF1 = Weight unit conversion factor (1E - 6 g/ μ g) t = Postapplication Day on which exposure is being assessed, a ssumed to be day zero MOE = Short Term Oral NOAEL (8.2 mkd) /[Daily Oral Dose (mg/kg/ day) . MOEs are reported to two significant figures Aggregate Oral and Dermal MOE = Short Term Oral NOAEL /Daily Oral Dose (all incidental oral) + Daily Dermal Dose 6 .0 AGGREGATE EXPOSURE AND RISK ASSESSMENT Inaccordance with the FQPA, when there are potential residential exposures to aP age 30 of 53

pesticide, aggregate risk assessment must consider exposures from three major routes: oral, dermal, and inhalation. There are three sources for these ty pes of exposures: food, drinking water, and residential 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 themse lves can be aggregated. When aggregating exposures and risks from various sources, HED considers both the route and duration of exposure. 6 .1 Acute & Chronic Aggregate Risk Acute and chronic aggregate expos ures include food plus drinking water exposures. Acute and chronic aggregate risks are not of concern. 6 .2 Short - Term Aggregate Risk To assess short -- term aggregate risk likely to result from the new and existing uses, HED combined average food and w ater exposure values with estimates of residential exposure for adult handlers and with residential exposure for adult and child post - application activities Tables 16 and 17 . Exposure for adult and child golfers was used to aggregate adult post - applicatio n 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. Table 16 . 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 yrs 0.0038 0.00 73 700 1 Residential Exposure = [Dermal exposure + Inhalation Exp osure]. 2 Aggregate MOE = [NOAEL ( 8.2 mg/kg/day  (Avg Food & Water Exposure + Residential Handler Exposure)] Table 17 . Short - Term

Aggregate Risk Calculations - Residential Post - Application Exposure LOC =100 Population Subgroup Average Food & Water E xposure mg /kg/day Residential Exposure 1 mg /kg/day Aggregate MOE (food and residential) 2 Females 13 - 49 yrs 0.0038 0.00 135 1600 Children 6 - 12 yrs 0.0059 0.00 194 1 000 Children 1 - 2 yrs 0.0093 0.0009 800 Children 3 - 5 yrs 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.P age 31 of 53

7 .0 OCCUPATIONAL EXPOSURE AND RISK 7 .1 Exposure Scenari os O ccupational handler and post - application exposure scenarios were assessed for th e risk assessment of the proposed new use s on cotton seed and turf . 7 .1.1 Handler Exposure Scenarios 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. 7 .1.1.1 Application to Turf

- Mixing and L oading a liquid formulation for groundboom application

- Applying a liqu id formulation with groundboom equipment.

- Mixing, L oading, and Applying a liquid fo rmulation with handgun spraying equipment. 7 .1.1.2 Seed Treatment

- L oading/Appl yi ng f

lowable concentrate for s eed treatment

- Sewing f

lowable concentrate

- Bagging f

lowable concentrate

- Multiple Activities with f lowable concentrate 7 .1.2 Post - Application Exposure Scenarios The following post - application exposure scenarios were assessed for proposed new acibenzolar - S - methyl uses. 7 .1.2.1 Turf Ap plication

- Mowing

- Hand weeding, transplanting

- Golfing 7 .1.2.2 Seed Treatment

- Seed Planting - Flowable concentrate for Seed treatment applications {single layer clothing with gloves).P age 32 of 53

7 .2 Occupational Exposure Data and Assumptions 7 .2.1 E xposure Data Maximum application rates for all of the exposure scenarios assessed are based on information provided in the acibenzolar - S - methyl labels for the proposed new uses . T he maximum single application rate for turf is 0. 0 22 lbs ai per acre . The maximum application rate for seed treatment is 9.9 x 10 - 6 lb ai/lb seed. D ata from the Pesticide Handlers Exposure Database (PHED) was used to assess handler and post - application exposures in the absence of chemical - specific data . The transfer coefficie nts used in the post - application exposure assessment are from an interim transfer coefficient guidance document developed by HED’s Science Advisory

Council for Exposure using proprietary data from the Agricultural Re - entry Task Force (ARTF) data base (SOP #3.1) . Post - application exposures were assessed for the proposed new uses. There are no compound specific data with which to estimate post application exposures to agricultural workers. Estimates of post - application re - entry exposure to agricultural wor

kers are based upon the EXPOSAC Standard Operating Procedures (SOPs) (3.1, Reference 4). This SOP lists a number of possible post - application agricultural activities for the proposed crop uses that m ight result in post - application. Post - application expos ure involving Commercial Seed treatment using acibenzolar - s - methyl consists of a “Planting Treated Seed” activity . HED’s SOP 14 is the data source for this “planting scenario” (which includes exposure to pesticide treated seed during both the loading of h oppers and driving the equipment). 7 .2.2 Exposure Assumptions The following standard exposure assumptions were used in estimating risks to workers from exposure to acibenzolar - S - methyl for the proposed new uses on onion and cucurbits. 7 .2.2.1 Han dler Exposures

• 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 by label review were used for all ty pes and methods of a

pplication.

• 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 for commercial turf applications (sod farm, athletic fields, commercial/industrial lawns)

- 40 acres treated per day for groundboom mix er/loader and applicator for golf course turf applications.

- 5 acres treated per day for handgun sprayer mixing, loading, and applying to turf areas .

- Seed treatment application frequency i s one event/application per season.

- T otal amount of seed treated is 160,000 lb/season for commercial treatment

- Total amount of seed treated is 3600 for on farm treatmentP age 33 of 53

7 .2.2. 2 Post Application Exposures

• 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 2500 cm 2 /hour (hand harvesting ) . • Exposure duration is 8 hours day . • Exposure is assumed to o ccur on the day of application (day

0) . • 72,000 lbs seed/season can be planted. 7 .3 Occupational Exposure and Risk Estimates 7 .3.1 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 toxi city endpoint was selected for these exposure routes . Exposure and risk estimates indicate MOEs are not of concern (MOEs > 100) at the maximum use rate for occupational handler activities for the proposed new uses. A summary of occupational handler expos ure and risk calculations, assumptions, and results is provided in Table s 18 and 19 . Table 18 : Short/Interm ediate - Term Occupational Handler Exposure and Risk for Acibenzolar - S - Methyl Exposure Scenario 1 Crop or Target App Rate (lb ai/A) 2 Acres

Treated Daily Unit Exposure 3 Dose (m g/kg/Day) 4,5 MOEs 6

Derm al (m g/lb ai) Inhalation

(μg/lb ai) Derm al Inhalation Combined Derm al + Inhalation Mixer/Loader Groundboom (Open Cab) C om mercial/ Industrial L aw

ns /Turf 0.022 80 2.9 1.2 0.0 340 3.5E - 06 240 Golf C o urse Turf 40 0. 0170 1.7E - 06 480 Mixer/Loader/Applicator Handgun Sprayer Commercial/

Industrial Lawns/Turf Golf Course Turf 0.022 5 0.5 1. 9 0.0004 3.5E - 06 22000 Applicator Groundboom (Open Cab) Commercial/

Industrial Lawns/Turf 0.022 80 0.014 0.000 74 0.000 16 2.0E - 0 5 44000 Golf course Turf 40 0.0000 82 1.1E - 0 5 88000 1 Use patterns are from the proposed labels 2 Application rates are based on maximum values based on proposed label. Most application rates upon which the analysis is based are pr esented as lb ai/A. In some cases, the application rate is based on applying a solution at concentrations specified by the label (i.e., presented as lb ai/gallon). 3 Baseline unit exposures. Values are reported in the PHED Surrogate Exposure Guide date d August 1998 or are from data submitted by the Outdoor Residential Exposure Task Force dated May 2000.P age 34 of 53 4 Inhalation dose (mg/kg/day) = [unit exposure (ug/lb ai) * 0.001 mg/ 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 we ight ( 6 0 kg). 5 Dermal Dose (mg/kg/day) = [unit exposure ( mg/lb ai) * Application rate (lb ai/acre or lb ai/gallon) * Daily area treated/amount handled (acres or gallons) * D ermal Absorption Factor (10%) ] / Body weight ( 6 0 kg). 6 Aggregate MOE = NOAEL (8.2 mkd)/Daily Inhalation Dose + Daily Dermal Dose Table 19 . Commercial Seed Treatment Handler Exposure and Risk for Acibenzolar - S - Methyl Exposure Scenario a Derm al Exp. b (m g/lb ai) Inhl.

Exp. c (ug/lb ai) Crop App. Rate

(lb ai/lb seed) Am nt.

Treated d (lbs. seed) Derm al

Dose e (m g/kg) Inhl.

Dose f (m g/kg) Combined

Dose g (m g/kg) M0E h Loader/ Applicator Single Layer w/ gloves 0.023 0.34 Cotton Seed 0.0000099 160,000 0.000 24 0.000009 0.000 3 4 33000

Sewer Single Layer w/no gloves 0.0062 0.23 Cotton Seed 0.0000099 160,000 0.0000 7 0.0000061 0.0.0000 7 12000

Bagger Single Layer w/ no gloves 0.0091 0.16 Cotton Seed 0.0000099 160,000 0.000 1 0.0000042 0.000 1 81000

M ultiple Activities Single Layer w/ gloves 0.042 1.6 Cotton Seed 0.0000099 160,000 0.000 4 0.000042 0.000 4 16000

On Farm Treatm ent Single Layer w/ gloves 12.6 1.2 Cotton Seed 0.0000099 3600 0.00 30 0.0000007 0.00 3 2700 a. PPE (Personal Protection Equipment) b. Dermal Unit Exposure is characterized with existing table. c. I nhalation Exposure is based on a Baseline exposure scenario (no respiratory protection). d. Amount/Seed treated values are based on label specific information (EPA Reg. No. 100 - 889 ) e. Dermal dose (mg/kg/event) = [unit dermal exposure (mg/lb ai) * dermal absorption (0.10) * application rate (lb ai/lb seed) * Amount treated / body weight (60 kg)]. f. Inhalation dose (mg/kg/event) = [unit exposure (kg/lb ai) * (1mg/1000 kg) conversion * appl. rate (lb ai/lbs seed) * Amount treated / body weight (60 kg)]. g. Combined event dose = event dermal dose + event inhalation dose. h. MOE = NOAEL

(mg/kg/d) / combined event dose. UF = 100. I. Gloves used for loading only. 7 .3.2 Post - Applicat ion 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 activiti es for the proposed new uses . A summary of post application exposure and risk calculations , assumptions, and results is provided in Table 20 and 21 .P age 35 of 53 1 Maximum application rate from proposed label 2 DAT = Day s after tr eatment needed to reach the LOC of 100; DAT 0 = the day of treatment/ assumed to be approx. 12 hours. 3 TTR (μg/cm 2 ) = Turf Transferable Residues corresponding to DAT, based on 20% of application rate. 4 TC (cm 2 /hr) = Transfer Coefficients and as sociated activities from ExpoSAC Policy Memo #003.1, 8/17/2000. 5 MOE = MOE on the corresponding DAT. MOE = NOAEL / Daily Dose. Daily Dose = [(TTR x TC x 10% 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 Table 21 . Seed Planter Exposure and Risk for Acibenzolar - S - Methyl Exposure Scenario a Derm al Exp. b (m g/lb ai) Inhl.

Exp. c (ug/lb ai) Crop App.

Rate (lb ai/lb seed) Am nt.

Treated d (lbs. se ed) Derm al

Dose e (m g/kg) Inhl.

Dose f (m g/kg) Combined

Dose g (m g/kg) M0E h Seed Planter I Single Layer w/ no gloves 0.25 3.4 Cotton Seed 0.0000099 3600 0.000 06 0.00009 0.000 15 55000 a. PPE (Personal Protection Equipment) b. Dermal Unit Exposure is charact erized with existing table. c. I nhalation Exposure is based on a Baseline exposure scenario (no respiratory protection). d. Amount/Seed treated values are based on label specific information (EPA Reg. No. 100 - 889) e. Dermal dose (mg/kg/event) = [unit der mal exposure (mg/lb ai) * dermal absorption (0.10) * application rate (lb ai/lb seed) * Amount treated / body weight (60 kg)]. f. Inhalation dose (mg/kg/event) = [unit exposure (kg/lb ai) * (1mg/1000 kg) conversion * appl. rate (lb ai/lbs seed) * Amount tr eated / body weight (60 kg)]. g. Combined event dose = event dermal dose + event inhalation dose. h. MOE = NOAEL

(mg/kg/d) / combined event dose. UF = 100. I. Gloves used for loading only. 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 determine 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 mechanis m of toxicity , EPA has not made a common mechanism of toxicity finding as to acibenzolar - S - methyl and any other substances and, acibenzolar - S - methyl does not appear to produce a toxic metabolite produced by

other substances which have tolerances in the U . S. For the purposes of this tolerance reassessment action, therefore, EPA has not assumed that acibenzolar - S - methyl has a common mechanism of toxicity with other substances. For information regarding EPA’s efforts Table 20 . Sum ma ry of Estim ated Occupational Post - application MOEs for Turf Crop Application

Rate (lb ai/A) 1 DAT 2 TTR 3

(μg/cm 2 ) TC 4

(cm 2 /hr) Activity 4 Short - /Int Term MOE 5

Turf 0.022 0 0.012 500 Mowing 32500 16500 Hand weeding, transplanting 1000 500 Golfing 32500Page 36 of 53

to determine which chemicals have a c ommon mechanism of toxicity and to evaluate the cumulative effects of such chemicals, see the policy statements released by

EPA’s OPP concerning common mechanism determinations and procedures for cumulating effects from substances found to have a common me chanism on EPA’s website at http://www.epa.gov/fedrgstr/EPA_PEST/2002/January /Day _16/

. 9 .0 DATA NEEDS 9 . 1 Toxicology Data Requirements

- Immunotoxicity (GLN 870.7 8 00)

- Battery

of mutagenicity

studies performed on technical - grade acibenzolar - S - methyl prepared by

the thi azole production process.

- Ames assay

using the technical - grade acibenzolar - S - methyl prepared by the thi azole production process - includ ing both pl ate incorporation and pre - incubation. (GLN 870.5100, 870.5300, 870.375, 870.900)Page 37 of 53

APPENDI CES 1.0 TOXICOLOGY DATA SUMMARY 1.1 Guideline Data Requirements Table 1. Guideline Data Requirem ents Test Technical Required Satisfied 870.1100 Acute Ora l Toxicity ................................ ....................... 870.1200 Acute Dermal Toxicity ................................ .................. 870.1300 Acute Inhalation Toxicity ................................ .............. 870.2400 Primary Eye Irritation ................................ .................... 870.2500 Primary Dermal Irritation ................................ .............. 870.2600 Dermal Sensitization ................................ ...................... yes yes yes yes yes yes yes

yes yes yes yes yes 870.3100 Oral Subchronic (rodent) ................................ ............... 870.3150 Oral Subchronic (nonrodent) ................................ ......... 870.3200 21 - Day Dermal ................................ .............................. 870.3250 90 - Day Dermal ................................ .............................. 870.3465 90 - Day Inhalation ................................ .......................... yes yes yes no yes y es

yes yes

- y es 870.3700a Developmental Toxicity (rodent) ................................ ... 870.3700b Development al Toxicity (nonrodent) ............................. 870.3800 Reproduction ................................ ................................ . yes yes yes yes

yes yes 870.4100a Chronic Toxicity (rodent) ................................ .............. 870.4100b Chronic Toxicity (nonrodent) ................................ ........ 870.4200a Oncogenicity (rat) ................................ .......................... 870.4200b Oncogenicity (mouse) ................................ .................... 870.4300 Chronic/Oncogenicity ................................ .................... yes yes yes yes yes yes

yes yes yes yes 870.5100 Mutagenicity

— Gene Mutation - bacterial ..................... 870.5300 Mutagenicity

— Gene Mutation - mammalian ................ 870.5375 Mutagenicity

— Structural Chromosomal Aberrations ... 870.5900 Mutagenicity

— Other Genotoxic Effects ....................... yes yes yes yes y es

yes yes yes 870.6100a Acute Delayed Neurotox. (hen) ................................ ..... 870.6100b 90 - Day Neurotoxicity (hen) ................................ ........... 870.6200a Acute Neurotox. Screening Battery (rat) ....................... 870.6200b 90 - Day Neuro. Screening Battery (rat) .......................... 870.6300 Develop. Neuro ................................ ............................. no no yes yes n o -

- yes yes

- 870.7485 General Metabolism ................................ ...................... 870.7600 Dermal Penetration ................................ ........................ 870.7800 Immunotoxicity ................................ .............................. yes

- Yes yes

- noPage 38 of 53

1.2 Toxic ity Profiles Table 2 Acute Toxicity Profile Acibenzolar - S - me thyl Acibenzolar - S - me thyl Guideline No./ Study T ype 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 4 4014220 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 I V 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 a nd 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 w eight, 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/d ay 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 w eight and body weight gain, increased spleen we ight and microsco pic changes in the spleenPage 39 of 53

Table 3 Subchronic, Chronic a nd 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) Acc eptable/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

– n ot 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 Mat erna l NOAEL = 50 mg/kg/day LOAEL = 300 mg/kg/day based on mortality, clinical signs of toxicity, decreased maternal body weight and food consumption. Developmenta l NOAEL = 300 mg/kg/day LOAEL = 600 mg/kg/day based on a marginal increase in vertebral anomal ies. 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 Mat ernal 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/ Systemic 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 40 of 53

Table 3 Subchronic, Chronic a nd 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 pending 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 we ights, 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 we ights, body weight gain and food efficiency, mild hemolytic anemia, and in creased 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 (isomer 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 5000  g/plate and less.Page 41 of 53

Table 3 Subchronic, Chronic a nd 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 - 3 23060 (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. Compound tested to cytotoxic concentrations. 870.5375 In vitro mammalian chromosome aberrati on (CHO cells) 44014245 (1993) Acceptable/guideline 7.5 to 60  g/ml ( - S - 9 and +S 9) Suggestive of clastogenicity in 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. I ncrease in polyploid cells at 30 and 60  g/mL at the 42 hour harvest time both with and without S 9. Evidence of cell 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) A cceptable/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 major 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 (19 92) 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 h ematology parameters accompanied by increased spleen weights.Page 42 of 53

Table 3 Subchronic, Chronic a nd Other Toxicity Profile for Acibenzolar - S - me thyl Guideline No. Study Type MRID No. (year) Classification Dose Levels Results 28 - Day oral gavage 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, decreased 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: 3 0.6 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 200 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 (bone 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 when 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 when dams were treated on GD 6 - 7 and 8 - 9. Dermal developmental toxicity rats 44014240 (1994) Accept able/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 w eights during lactation.Page 43 of 53

2.0 Studies Reviewed for Ethical Conduct The PHED Task Force, 1995. The Pesticide Handlers Exposure Database, Version 1.1. Task Force me mbers Health Canada, U.S. Environmental Protection Agency , and the National Agricultural Chemicals Association, released February , 1995. Agricultural Re - entry

Task Force (ARTF) data base (SOP #3.1)Page 44 of 53

3 .0 ADDITIONAL TOXICOLOGY STUDIES 870.3700 Developm ental Toxicity Ina non - guideline developmental toxicity study (MRID 44014238), twelve presumed pregnant Tif:RAT f(SPF) (hy

brids of RTT / 1 x RI I/2) rats per group were administered CGA 245704 Technical (97.9%; Batch No. P.303011) in a 0.5% aqueous solution of sodium carboxy methylcellulose by

gavage at a dose of 300 mg/kg/day . Groups 2 - 7 were administered test article on gestation days (GD) 6 - 15,6 - 7, 8 - 9, 10 - 11 , 12 - 13, or 14 - 15, respectively . Group 1 consisted of twelve animals which served as controls and were treated with 0.5% sodium carboxy methylcellulose on GD 6 - 15. On GD 21, dams were sacrificed, subjected to gross necropsy , 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 dams 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 days starting on GD 13 and was not observed in controls or in animals in Group 7. No other treatment - related clinical signs of toxicity

were observed. No statistically significant differences in absolute body weights or body

weight changes occurred at any 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 statistica lly 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 significant, 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 early 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, respectively . In one Group 2 litter, generalized 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 greatly

reduced body

weights. Generalized edema was also seen in one fetus 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 pronounc ed maternal and developmental toxicity occurred when dams were treated 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. Ina non - guideline devel opmental toxicity study (MRID 44014239), eight presumed pregnant Tif: RAT f (SPF) (hy

brids of RI I/ 1 x RII2) 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 days (GD) 6 - 7, 8 - 9, 10 - 11, 12 - 13, or 14 - 15, respectively . Group 1 consisted of eight animals which served as controls and were treated with 0.5% carboxy methylceliulose (vehicle) on GD 6 - 15. On GD 21, dams were sacrificed, subject ed toPage 45 of 53

gross necropsy , 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 necropsy findings including mottled ki dney and thy mus. One Group 4 dam delivered on GD 20 and was sacrificed. 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 any

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 bo dy weights, or fetal sex ratios. Group 2 had a slightly greater number of early

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 pl acentae 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, respectively . 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 any

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. Pronounced 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 Penetration Ina 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 carboxy

methylcellulose (0.5% w/w) in 0.1% aqueous Polysorbate 80, by dermal application at doses of 0 (vehicle), 10, 100, or 500 mg/kg/day o n gestation 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 litte r 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 any treatment level. Gestational body weights, body

weight changes, and food consumption were not statistically

significantly

different for any

treatment level throughout the gestational period. Therefore, the maternal toxicity NOAEL is 500 mg/kg/day based on no adverse mate rnal effects 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 biologically relevant statisticallyPage 46 of 53

significant differences in the incidence rates of any external, visceral, or skeletal malformations/ anomalies/variations were observed 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 phal anx 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 biologically significant. Therefore, the developmental toxicity NOAE L is 500 mgfkglday based on no adverse d evelopmenta l effects at the highest dose administered. The developmental toxicity L OAEL was >500 mg/kg/day . Ina 28 - day

repeated dose dermal toxicity study (MRID 44014233) , groups of 5 fe males Tif: RAI f (SPF) albino rats were treated with acibenzolar - S - m ethyl technical in 0.5% (w/v) carboxy methyl cellulose/0.1% (w/v) aqueous polysobate 80 at does of 0, 1, 10, 100 or 1000 mg/kg/day . Animals were treated by 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 necropsy . The sy stemic and dermal NOAEL is the limit do se of 1000 mg/kg/day . A dermal and sy stemic L OAEL were not identified. 870.3800 Reproduction Ina 1 - generation range - finding reproduction study in rats (Nonguideline study , MRID 44014241 ) CGA 245704 Technical (97.9% a.i.; Batch No. P.303011) was admin istered to groups of 15 male and 15 female Tif: RAT f (SPF), hy brids of RTTI1 x P11/2 (Spra gu e - Dawley derived) rats at dietary concentrations of 0, 2000, 4000, or 8000 ppm for one generation (MRID 44681301). In this range - finding study , F0 adults were 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 , respectively , for F0 females. Males were terminated at the end of matin g 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 necropsied. The only clinical sign of toxicity

in the parental animals was perineal hemorrhage during late gestation in five high - dose females. Gross necropsy was unremarkable. Absolute body weights, body

weight gains, and food consumption values for the low - and mid - dose males and low - dose females were similar to the controls throughout the study . High - dose males had significantly (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 significantly (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 days 1 - 8. Reduced body weight gains in the high - dose males and females and mid - dose fema les correlated with significantly

(p > 0.05 or 0.01) reduced food consumption by these treated groups during study day s 1 - 8. L ower body

weights, body

weight gains, and food consumption continued for the mid -Page 47 of 53

and high - dose females during gestation and lactation. Body weights of the high - dose group were significantly

(p > 0.05 or 0.01) lower than the controls throughout gestation with overall weight gain by the high - dose females during gestation 27% (p > 0.01) of the control group level. Mid dose females had significantly (p > 0.05 or 0.01) lower body weights than the controls on GD 21 and on lactation days 7, 14, and 21. Weight gains by the mid - dose group during gestation and lactation were 92% and 83%, respectively of the control group levels. Food consumpti on by the high - dose group was significantly (p > 0.05 or 0.01) less than the controls throughout gestation. Food consumption by

the mid - dose group was significantly (p > 0.01) less than the controls throughout lactation. Therefore, the LOAEL for parental t oxicity 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 i mplantation 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) . Li ve 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 significantly

(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 body we ight gains from postnatal day 4 through postnatal day

28. Recovery

of body weight gains occurred during the postweaning intervals of days 28 - 35 and 35 - 42. Therefore, the LOAEL for developmental toxicity

is 4000 ppm based on reduced pup body weight gains an d lower pup body weights during lactation and early postweaning. The developmental toxicity NOAEL is 2000 ppm. 870.6200b Subchronic Neurotoxicity Ina 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 day s. Neurobehavioral assessment (functio nal 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 tissues from the perfused animals in the control and 8000 ppm groups were subjected to histopathological evaluation of brain and peripheral nervous sy stem tissues. Positive control data were not provided. No compound related effects were observed in mortality , clin ical signs of toxicity , FOB, motor activity , or gross or neuropathology . At 8000 ppm, body weights were generally

decreased throughout the study in the males (decr 11 - 14%, not statistically significant) and females (decr 8 - 13%, attaining statistical signif

icance at Weeks 2 - 5, 7 and 8, and 11 - 13). Also at this dose, cumulative bodyPage 48 of 53

weight gains were decreased throughout the study in the males (decr 19 - 35%, attaining statistical significance at Weeks 2 - 4 and 6 - 9), and in the females (decr 24 - 40%). A decreas ing trend (p<0.01) in body

weight and body

weight gain was also generally noted in both sexes at this dose. These decreases in body weight gain corresponded with decreases in food consumption (g/animal/day

) which was sporadically

decreased in the 8000 ppm males (decr 9 - 21% at Weeks 1 - 3, 5 - 7, and 11, but only attaining statistical significance during Week 1), and females (decr 14 - 18%, p<0.01 at Weeks 1 and 3). A decreasing trend (p<0.01) in food consumption was also generally 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 consumption . The NOAEL is 2000 p pm (equivalent to 126/143 mg/kg/day , M/F ) . T his study is classified as acceptable/nonguideline pending submission of positive control data for neuropathological and FOB assessments and does not satisfy the guideline requirement (OPPTS 870.6200b) for a sub chronic neurotoxicity study in rats. The study

may be upgraded pending submission of acceptable positive control data. 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 RID 44014246) 3. Chromosome aberration, Chinese hamster ovary (CHO) cells in culture ( MRID 4 4014245) 4. Unscheduled DNA sy nthesis assay , primary

rat hepatocy

tes in culture (MRID 44014248) 5. Invivo cy

togenetics, micronucleus assay in mice (MRID 44014244) Results in all five studies were negative for genotoxicity . The five studies on the techn ical 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 by- 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 gen e mutation, S. ty phimuriumfE. Coli; Test Material: CGA - 323060 (M R lD 44537026). This study

was negativePage 49 of 53

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 ye t been submitted to the Agency . Trace levels of this isomer (<0.1%) were found in some batches of technical grade acibenzolar- S niethyl produced via the 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. Presumably , the same isomer does not occur in the old production process which produced the test material (lot/batch # P.303011) that wa s used for virtually all the toxicity

testing on acibenzolar - S - methyl up to this point in time.Page 50 of 53

4.0 TOXICOLOGY STUDY REFERENCES 44014227 Bachmann, M. (1992) CGA - 245704 Technical: 28 - Day Range Finding Study

in Rats: (Administration in Food): (Final Report): Lab Project Number: 923060. Unpublished study prepared by

Ciba - Geigy

Ltd. 221 p. 44014228 Fankhauser, H. (1993) CGA - 245704 Technical: 3 - Month Range Finding Toxicity Study in Mice: (Administration in Food): (Final Report): Lab Project Number: 923081: 3 31018. Unpublished study

prepared by

Ciba - Geigy

Ltd. 214 p. 44014229 Fankhauser, H. (1993) CGA - 245704 Technical: 28 - Day Oral Cumulative Toxicity Study in Rats (Gavage): (Final Report): Lab Project Number: 923137: 349064. Unpublished study prepared by

Cib a - Geigy

Ltd. 276 p. 44014230 Bachmann, M. (1993) CGA - 245704 Technical: 3 - Month Oral Toxicity Study in Rats: (Administration in Food): (Final Report): Lab Project Number: 923082: 338433. Unpublished study prepared by

Ciba - Geigy

Ltd. 404 p. 44014231 Altm ann, B. (1994) CGA - 245704 Technical: 28 - Day Range Finding Toxicity

Study in Beagle Dogs: (Final Report): Lab Project Number: 923130. Unpublished study prepared by

Ciba Geigy Ltd. 238 p. 44014232 Altmann, B. (1994) CGA - 245704 Technical: 90 - Day Subchronic Oral Toxicity Study in Beagle Dogs: (Final Report): Lab Project Number: 923138. Unpublished study prepared by Ciba - Geigy

Ltd. 417 p. 44014233 Hagemann, C. (1994) CGA - 245704 Technical: 28 - Day Repeated Dose Dermal Toxicity Study in the Rat: (Final Report): Lab Project Number: 923135: 353867. Unpublished study prepared by Ciba - Geigy

Ltd. 248 p. 45713601 Classen, W. (1997) 90 - Day Subchronic Neurotoxicity Study in Rats: CGA - 245704 Technical: Final Report: Lab Project Number: 963028: 568 - 96: CH - 4002. Unpublis hed study prepared by Novartis Crop Protection, Inc. 259 p. {OPPTS 870.6200} 44014234 Altmann, B. (1996) CGA - 245704 Technical: 12 - Month Chronic Oral Toxicity Study in Beagle Dogs: (Final Report): Lab Project Number: 923149. Unpublished study prepared by Ciba Geigy Ltd. 574 p. 44014243 Fankhauser, H. (1996) CGA - 245704 Technical: 24 - Month Carcinogenicity and Chronic Toxicity Study in Rats: (Final Report): Lab Project Number: 923151. Unpublished study prepared by

Ciba - Geigy

Ltd. 1587 p. 44014235 Fankhaus er, H. (1996) CGA - 245704 Technical: 18 - Month Oncogenicity Study in Mice: (Final Report): Lab Project Number: 923150. Unpublished study prepared by

Ciba - Geigy

Ltd. 1024 p. 44014243 Fankhauser, H. (1996) CGA - 245704 Technical: 24 - Month Carcinogenicity and C hronic Toxicity Study in Rats: (Final Report): Lab Project Number: 923151. Unpublished study prepared by

Ciba - Geigy

Ltd. 1587 p. 44014236 Marty , J. (1994) CGA - 245704 Technical: Rat Oral Teratogenicity : (Final Report): Lab Project

Number: 923139: 349097. Unpublished study prepared by

Ciba - Geigy

Ltd. 407 p. 44014237 Marty , J. (1994) CGA - 245704 Technical: Rabbit Oral Teratogenicity : (Final Report): LabPage 51 of 53 Project Number: 923140: 360382. Unpublished study prepared by

Ciba - Geigy

Ltd. 266 p. 44014238 Khalil, S . (1994) CGA - 245704 Technical: Range Finding Rat Oral Teratogenicity : (Non Standard Study): (Final Report): Lab Project Number: 943067. Unpublished study prepared by Ciba - Geigy

Ltd. 135 p. 44014239 Khalil, S. (1994) CGA - 245704 Technical: Range Finding Rat Oral Teratogenicity : (Non Standard Study): (Final Report): Lab Project Number: 943004. Unpublished study prepared by Ciba - Geigy

Ltd. 118 p. 44014240 Khalil, S. (1994) CGA - 245704 Technical: Rat Dermal Teratogenicity : (Final Report): Lab

Project Number: 943001: 370440. Unpublished study prepared by

Ciba - Geigy

Ltd. 381 p. 45089701 Miyamoto, M. (1998) Teratology Study of Acibenzolar - S - methyl in Rats: Final Report: Lab Project Number: AN98025: 1197 - 98. Unpublished study prepared by

Ina Research Inc. 131 p. {OPPTS 870.3700} 44014241 Khalil, S. (1995) CGA - 245704 Technical: Two - Generation Reproduction Study in Rats: (Final Report): Lab Project Number: 923152. Unpublished study prepared by

Ciba - Geigy

Ltd. 1628 p. 44681301 Marty , J. (1993) CGA - 245704 Technic al: Rangefinding Rat Dietary Reproduction Study : Lab Project Number: 923146: 634 - 93. Unpublished study prepared by

Novartis Crop Protection, Inc. 65 p. 44014244 Hertner, T. (1993) CGA - 245704 Technical: Micronucleus Test: Mouse (OECD Conform) in

vivo Study: Lab Project Number: 923143. Unpublished study prepared by

Ciba - Geigy

Ltd. 40 p. 44014245 Hertner, T. (1993) CGA - 245704 Technical: Cy togenetic Test on Chinese Hamster Cells in vitro (EC - Conform): Lab Project Number: 923142. Unpublished study prepared b y Ciba - Geigy Ltd. 45 p. 44014246 Geleick, D. (1993) CGA - 245704 Technical: Gene Mutation Test with Chinese Hamster Cells

V79 in vitro: Lab Project Number: 923141. Unpublished study prepared by

Ciba - Geigy

Ltd. 53 p. 44014247 Hertner, T. (1993) CGA - 245704 Technical: Salmonella and Escerichia/Liver Microsome Test:

Gene Mutation Test: Lab Project Number: 923145. Unpublished study prepared by

Ciba - Geigy Ltd. 67 p. 44014248 Hertner, T. (1993) CGA - 245704 Technical: Autoradiographic DNA Repair Test on Rat

Hepatocy tes (OECD Conform) in vitro: Lab Project Number: 923144. Unpublished study prepared by

Ciba - Geigy

Ltd. 86 p. 44537024 Deparade, E. (1998) Salmonella and Escherichia/Mammalian - Microsome Mutagenicity Test: NOA 419191 Technical: Lab Project Number: 9730 97: 324 - 98: A98013. Unpublished study prepared by

Novartis Crop Protection, AG. 75 p. 44537025 Deparade, E. (1998) Salmonella and Escherichia/Mammalian - Microsome Mutagenicity Test: CGA - 362020 Technical: Lab Project Number: 973079: 325 - 98. Unpublished stu dy prepared by Novartis Crop Protection, AG. 74 p. 44537026 Ogorek, B. (1997) Salmonella and Escherichia/Mammalian - Microsome Mutagenicity Test: CGA - 323060 Technical: Lab Project Number: 963125: 761 - 97: A960043. Unpublished studyPage 52 of 53 prepared by

Novartis Crop Protection, AG. 74 p. 45434101 Deparade, E. (1998) Salmonella and Esherichia/Mammalian - Microsome Mutagenicity Test: CGA - 245704: Final Report: Lab Project Number: 983053: 1240 - 98. Unpublished study prepared by

Novartis CropProtection AG. 71 p. {OPPTS 870 .5100} 44014250 Molitor, E. (1995) The Metabolism of (U - (carbon 14))Phenyl CGA - 245704 in the Rat: Lab

Project Number: 13/94: 016AM03. Unpublished study prepared by

Ciba - Geigy

Ltd. 197 p.Page 53 of 53

5.0 CHEMICAL NAMES AND STRUCTURES OF METABOLITES

N N S S C H 3 O

Acibenzolar N N S C O O H

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 wat er