Page 1 of 94 UNITED STATES ENVIRONMENTAL PROTECTION AGENCY WASHINGTON, D.C. 20460

OFFICE OF CHEMICAL SAFETY
AND POLLUTION PREVENTIO N

MEMORANDUM

Date: 9/2/10

SUBJECT: Cyflufenamid : Human Health Risk Assessment for Proposed Uses on Cucurbit Vegetables (Crop Group 9), Pome Fruit (Cr op Group 11), Small Fruit Vine Climbing Except Fuzzy Kiwifruit (Subgroup 13-07F), and Lo w Growing Berry (Subgroup 13-07G), except Cranberry.

PC Code: 555550 DP Barcode: : D360825
Decision No.: 403833 Registration No.: 8033-RNG
Petition No.: 8F7488 Regulatory Action: Section 3 Registration Risk Assessment Type: Single Chemical/Aggregate Case No.: NA

TXR No.: NA CAS No.: 180409-60-3
MRID No.: NA 40 CFR: 180.xxx (pending)

FROM: Nancy Dodd, Risk Assessor Risk Assessment Branch III (RAB3) Health Effects Division (HED) (7509P)

AND Sheila Healy, Ph.D., Toxicologist Meheret Negussie, Chemist Cassi L.Walls, Ph.D., Senior Biologist RAB3/HED (7509P)

THROUGH: Paula Deschamp, Branch Chief RAB3/HED (7509P)

AND Christina Swartz, Branch Chief PV Shah, Toxicologist

Risk Assessment Review Committee

TO: Mark Suarez/Samantha Hulkower, Risk Management Team #11

Insecticide Branch Registration Division (7505P)Page 2 of 94 Table of Contents

1.0 Executive Summary .......................................................................................................... 4

2.0 Regulatory Conclusions and Recommendations ............................................................ 7

2.2.1 Enforcement Analytical Method ............................................................................. 9

2.2.2 International Harmonization ................................................................................... 9

2.2.3 Recommended Tolerances ........................................................................................ 9

2.2.4 Revisions to Petitioned-For Tolerances ................................................................ 10

2.3 Label Recommendations ............................................................................................ 10

2.3.1 Recommendations from Residue Reviews ............................................................ 10

2.3.2 Recommendations for Worker Safety ................................................................... 10

2.3.3 Recommendations for Residential Safety ............................................................. 11

3.0 Ingredient Profile ............................................................................................................ 11

3.1 Structure and Nomenclature ..................................................................................... 11

3.2 Physical/Chemical Characteristics ............................................................................ 12

3.3 Summary of Existing/Proposed Uses ........................................................................ 12

4.0 Hazard Characterization and Dose-Response Assessment ......................................... 14

4.1 Toxicology Studies Available for Analysis ................................................................ 14

4.2 Absorption, Distribution, Metab olism, & Elimination (ADME) ............................ 15

4.2.1 Dermal Absorption ................................................................................................. 16

4.3 Toxicological Effects ................................................................................................... 17

4.4 Safety factor for Infants and Children (FQPA Safety Factor) ............................... 18

4.4.1 Completeness of the Toxicology Database ............................................................ 18

4.4.2 Evidence of Neurotoxicity ...................................................................................... 19

4.4.3 Evidence of Sensitivity/Susceptibility in the Developing or Young Animal ...... 20

4.4.4 Residual Uncertainty in the Exposure Database .................................................. 20

4.5 Toxicity Endpoint and Point of Departure Selections ............................................. 21

4.5.1 Dose-Response Assessment .................................................................................... 21

4.5.2 Recommendation for Combining Routes of Exposures for Risk Assessment ... 22

4.5.3 Classification of Carcinogenic Potential ............................................................... 22

4.5.4 Summary of Points of Departure and T oxicity Endpoints Used in Human Risk

Assessment

.............................................................................................................................. . 22

4.5.5 Endocrine Disruption ............................................................................................. 24

5.0 Dietary Exposure and Risk Assessment ........................................................................ 25

5.1 Metabolite/Degradate Residue Profile ...................................................................... 25

5.1.1 Summary of Plant and Animal Metabolism Studies ........................................... 25

5.1.2 Summary of Environmental Degradation ............................................................ 26

5.1.3 Comparison of Metabolic Pathways ...................................................................... 27

5.1.4 Residues of Concern Summary and Rationale ..................................................... 28

5.2 Food Residue Profile ................................................................................................... 31

5.3 Water Residue Profile ................................................................................................. 33

5.4 Dietary Risk Assessment ............................................................................................ 34

5.4.1 Description of Residue Data Used in Dietary Assessment .................................. 34

5.4.2 Percent Crop Treated Used in Dietary Assessment ............................................. 34

5.4.3 Acute Dietary Risk Assessment ............................................................................. 34Page 3 of 94 5.4.4 Chronic Dietary Risk Assessment ......................................................................... 35

5.4.5 Cancer Dietary Risk Assessment ........................................................................... 35

5.4.6 Summary Table ....................................................................................................... 35

6.0 Residential (Non-Occupational) Exposure/Risk Characterization ............................ 36

6.1 Residential Handler Exposure ................................................................................... 36

6.2 Post-Application Exposure ......................................................................................... 39

6.3 Combined Exposure .................................................................................................... 39

6.4 Spray Drift ................................................................................................................... 40

7.0 Aggregate Exposure/Risk Characterization ................................................................. 40

7.1 Acute Aggregate Risk ................................................................................................. 40

7.2 Short- and Intermediate-Term Aggregate Risk ....................................................... 40

7.3 Chronic Aggregate Risk ............................................................................................. 41

7.4 Cancer Aggregate Risk ............................................................................................... 41

8.0 Cumulative Exposure/Risk Characterization .............................................................. 42

9.0 Occupational Exposure/Risk Characterization ........................................................... 42

9.1 Short-/Intermediate-/Long-Term/Cancer Handler Risk ......................................... 42

9.2 Short-/Intermediate-/Long-Term/Cancer Post-Application Risk .......................... 47

10.0 Incidents ........................................................................................................................... 49

11.0 Environmental Justice Considerations ......................................................................... 49

12.0 References ........................................................................................................................ 50 Appendix A. Toxicology Prof ile and Executive Summaries .................................................. 51

A.1 Toxicology Data Requirements .................................................................................. 51

A.2 Toxicity Profiles .......................................................................................................... 52

A.3 Hazard Identification and Endpoint Selection ......................................................... 58

A.4 Executive Summaries .................................................................................................. 59 Appendix B. Metabolism Summary Table .............................................................................. 85 Appendix C. Review of Human Research ................................................................................ 92 Appendix D. International Residue Limit Status Sheet ......................................................... 93 Appendix E. Data Required as a Condition of Registration .................................................. 94Page 4 of 94 1.0 Executive Summary

Use Profile : Nippon Soda Company, Ltd. has proposed uses for cyflufenamid on cucurbit vegetables (Crop Group 9), pome fruit (Crop Group 11), small fruit vine climbing except fuzzy kiwifruit (Subgroup 13-07F), low growing berry (Subgroup 13-07G), except cranberry, and outdoor landscape and outdoor nursery ornamentals. The petitioner voluntarily withdrew the use on cranberry. Cyflufenamid is a fungicide which controls powdery mildews. Its mode of action

has not been determined.

Cyflufenamid is formulated as suspension con centrates, and applied as postemergence broadcast foliar sprays using ground or aer ial equipment. Application through any type of irrigation system is prohibited. A maximum of two applic ations can be made to agricultural crops and

ornamentals for a maximum of 0.022 lb ai/A/growing season (cucurbits), 0.044 lb ai/A/growing season (other agricultural crops), and 0.044 lb ai/A/y ear (ornamentals). For agricultural crops (as shown in Table 3.3), retreatment intervals (RTIs) are 7 or 14 days and preharvest intervals (PHIs) are 0, 3, or 14 days. Retreatment interval s for ornamentals are 14- 21 days. The restricted entry interval (RE I) is 12 hours.

These actions will be the first re gistrations in the U.S. Cyflufenamid is registered in Japan, Korea, and several European Union (EU) countries.

Human Health Risk Assessment:

Cyflufenamid has low acute toxicity via the oral , dermal and inhalation routes of exposure. Though slightly irritating to the eye, cyflufenamid is not a skin irritant or sensitizer. In the longer-term studies of the mamma lian toxicology database, the liver was the primary target organ of cyflufenamid toxicity. Across species, duration and gender, changes in weight, clinical chemistry and pathology indicated treatment-relate d perturbations in and adverse effects on liver

function. Treatment-related effects were also observed in the rat thyroid and kidney. Cyflufenamid-induced cardiotoxicity in the mouse and rat, myocar dial vacuolation and lipidosis

could be ascribed to the in vitro inhibition of carnitine palmitoyltransferase by cyflufenamid.

An acute endpoint attributable to a single dose was not identified. The chronic NOAEL of 4.4 mg/kg/day was based on decrea sed body weight gain, increased thyroid/parathyroid weight, increased liver weight and cen trilobular hepatocytic hypertrop hy in the combined chronic toxicity/carcinogenicity study in ra ts. No dermal hazard was identi fied. The incidental oral and inhalation NOAELs of 5 mg/kg/day are based on decreased body weight, body weight gain and food consumption in a prenatal de velopmental study in rabbits.

Cyflufenamid is classified as “likely to be car cinogenic to humans.” This was based on the presence of two tumor types in two species: thyroid follicular cell tumors in male rats and liver

tumors in male mice. There is no con cern for mutagenicity. The unit risk, Q 1 * , of cyflufenamid based upon male mouse liver combined ade noma and carcinoma tumor rates is 6.61 x 10 -3
(mg/kg/day) -1 in human equivalents.Page 5 of 94 Dietary Exposure (Food/Water) :

The residue chemistry data and environmental fate data are adequate to assess human exposure. The chronic and cancer dietary food exposure asse ssments are conservative. The assessments are based on reliable data and will not underestimate exposure/risk.

Chronic: A partially refined ch ronic dietary assessment was conducted. The assumptions of this dietary assessment included av erage field trial residues for all proposed crops and 100% crop treated (CT). Empirical or DEEMTM default pr ocessing factors were used. The ground water estimated drinking water concentration (EDWC) of 4.68 u g/L was incorporated directly into the dietary assessment (DP# 362954, R. Bohaty, 5/3/2010).

The non-cancer chronic dietary (f ood and drinking water) exposure to cyflufenamid is below HED’s level of concern [i.e., <100% of the chr onic Population Adjusted Dose (cPAD)] for the general U.S. population and all population subgroups . The chronic dietary exposure estimates are <1% of the cPAD for the general U.S. popula tion and 1% of the cPAD for all infants (<1 year old), the most highly exposed population subgroup.

Cancer: The cancer dietary (food and drinking water) exposure estimate for the general U.S. population is 9.5 x 10 -7 , which is below HED’s level of conc ern (generally in the range of 10 -6 ).

Residential Exposure :

Residential Handler: Residential short-term (1 - 30 days) dermal and inhalation exposures are expected from handler activities associated with the proposed ornamental uses. Since no hazard was identified for the dermal route of exposure, dermal risks were not assessed. HED used data

from ORETF and PHED Version 1.1 to assess residential handler exposures. Margins of Exposure (MOEs) greater than 100 for the inhala tion route are deemed adequate to protect residential cyflufenamid handlers. All inhalati on scenarios for residential handlers resulted in MOEs greater than the level of concern LOC (i.e., MOEs ≥ 100) and, therefore, are not of

concern. All residential handler scenarios re sulted in cancer risks which do not exceed HED’s level of concern (generally in the range of 10 -6 ) when wearing baseline clothing (e.g

. , shortsleeve shirt, short pants, and no gl oves). It should be noted that the proposed label states “Not for homeowner use”; however, HED assessed re sidential handler exposure activities for the ornamental uses because there is a potential th at homeowners could apply this product. HED recommends that the label language be revised to clarify the specific type of applicator and setting (i.e., “applications made only by license d pest control operators in commercial and residential settings”).Page 6 of 94 Residential Postapplication: Pote ntial residential postapplication expos ure to adults and children from ornamental applications are expected to be negligible for the following reasons: 1) Activities such as pruning/thinning ornamentals or pl aying in and around ornamentals wh en residues may be present on the day of the application ar e unlikely to co-occur; 2) if present, le af to skin residue transfer would be negligible because of the minimal frequency an d duration of contact; 3) children young enough to

exhibit hand-to-mouth behavior woul d not typically play in ornament al beds or trees. Based on the frequency of application and unlik ely potential for postapplication e xposure, residential postapplication risks were not quantitatively assessed; thus, there are no posta pplication residential risk concerns for this
use pattern.

Based on the Agency's current practices, a qua ntitative postapplication inhalation exposure assessment was not performed for cyflufenamid at this time. If new policies or procedures are put into place, the Agency may revisit the ne ed for a quantitative pos tapplication inhalation exposure assessment for cyflufenamid.

Aggregate Risk:

Acute: Acute exposures were not considered because an adverse effect attributable to a single

dose was not identified.

Chronic: Aggregate chronic risk is equivalent to chr onic dietary risk. Ther e is potential chronic exposure ( ≤ 1% of the cPAD) via the diet (food and drinki ng water) as determin ed in Section 5.4.

Short- and Intermediate Term: There is poten tial short- and intermediate-term exposure to cyflufenamid via the dietary pathway (which is considered bac kground exposure) and the residential pathway (which is considered the pr imary pathway). Since no dermal hazard was identified, the exposure pathways for adults include the oral (background) and inhalation (primary) routes. For children, the exposure path way includes only the or al (background) route since 1) no dermal hazard was identified; 2) or namentals are the only residential use; and 3) dermal, inhalation, and incidental oral expos ure to children from treated ornamentals is considered to be negligible. All aggregate MO E’s are greater than 100; therefore, aggregate exposure is not of concern.

Cancer: The aggregate cancer risk (f ood, water, and residential) is 9.7 x 10 -7

.

Occupational Exposure/Risk:

Agricultural Handlers: Occupa tional short-term (1 - 30 days) and intermediate-term (1 - 6 months) dermal and inhalation exposures are ex pected from cyflufenam id handler activities associated with both the proposed agricultura l and ornamental uses. Since no hazard was identified for the dermal route of exposure, non-cancer dermal risks were not assessed.

HED used data from the Pesticide Handlers Expo sure Database (PHED) Version 1.1 as presented in the PHED Surrogate Exposure Guide (8/98) to assess handler e xposures (HED Science Advisory Council for Exposure (ExpoSAC) Standa rd Operating Procedure (SOP) No. 7, dated 01/28/1999). A Margin of Exposure (MOE) greater than 100 for the inhalation route is deemedPage 7 of 94 adequate to protect occupational cyflufenamid handlers. All base line inhalation scenarios (i.e., no respiratory protection) for handl ers resulted in MOEs greater than the level of concern LOC (i.e., MOEs ≥ 100) and, therefore, are not of con cern. The maximum exposure (MOE = 38,000) occurred from mixing/loading liquid concen trates for aerial applications to pome fruit/grapes/strawberries at the maximum applicati on rate (0.022 lb ai/A). The cancer risks range

from 5.0 x 10 -9 to 1.2 x 10 -7 assuming the workers wear labe led Personal Protective Equipment (PPE) (long-sleeved shirt, long-pants, and chemical resistant gloves).

Agricultural Postapplication: Since no hazard was identified for the dermal route of exposure, occupational short- and intermediate-term non- cancer dermal risks were not assessed for postapplication activities. Based on the Agency's current practices, a qua ntitative postapplication inhalation exposure non-can cer assessment was not performed for cyflufenamid at this time.

Using surrogate data and default assumptions, th e cancer risks from agricultural postapplication activities range from 7.4 x 10 -8 to 4.1 x 10 -6 on day 0 (12 hours following application). Although cyflufenamid specific dislodgeable foliar resi dues (DFR) studies on grapes and ornamentals could be used to refine and mitigate the cancer risks for these scenarios, this assessment incorporates conservative parame ters such as maximum application rates and assuming workers are exposed to “Day 0” cyflufenamid residue levels for 250 days/y ear even though only two applications per ornamental type per year can be made with a retr eatment interval of 21 days.

Environmental Justice Considerations: Potential areas of environm ental justice concerns, to the extent possible, were considered in this human health risk assessment. Dietary and nondietary exposures were considered.

Review of Human Research: This risk assessment relies in pa rt on data from studies in which adult human subjects were intent ionally exposed to a pesticide or other chemical. These studies (listed in Appendix C) have been determined to require a review of thei r ethical conduct, and one of these studies is also subject to review by the Human Studies Review Board. All of the listed

studies have received the appropriate review.

2.0 Regulatory Conclusions and Recommendations

Provided revised Sections B (OPPTS 860.1200) and F (OPPTS 860.1550) are submitted, HED can recommend for a conditional registration and permanent tolerances for the proposed uses of cyflufenamid on pome fruit, cucurbits, and the fruit subgroups 13-07F and 13-07G, except cranberry. Additional data are needed as condi tions of registration as outlined in Section 2.1 below. The specific tolerance recommendati ons are discussed in Section 2.2, and label

modifications are discussed in Section 2.3.

Although not a requirement since there is a multi residue method, the Agency is requesting that the petitioner revise the liquid chromat ography/tandem mass spect rometric detection (LC/MS/MS) enforcement method (Method 070276) to provide two mass ion transitions for analysis of cyflufenamid.Page 8 of 94 2.1 Data Deficiencies/Conditions of Registration

The following must be provided prior to a tolerance being established or registration

allowed:

860.1200 Directions for Use A revised Section B is requested prohibiting th e addition of a spray adjuvant to the spray solution and including plant-back intervals (PBIs) of 30 days for small grains and leafy vegetables and 365 days for all other crops not on the label.

860.1550 Proposed Tolerances A revised Section F must be submitted to reflect the tolerances and correct commodity

definitions as listed in Table 2.2.4.

The following data are required as a condition of registration:

TOXICOLOGY

870.6200 Neurotoxicity Battery (Acute Studies) The acute neurotoxicity screening battery is required to fulfill current 40 CFR Part 158 data requirements.

RESIDUE CHEMISTRY

860.1300 Nature of the Residue – PlantsTo upgrade the submitted apple metabolism study (MRID 47620805), details of the inlife and analytical phases of the study al ong with quantitative raw data and sample storage information must be submitted.

The following data should be submitted with future uses:

RESIDUE CHEMISTRY

860.1300 Nature of the Residue – Plants If additional crops are proposed in the future, the ChemSAC recommends that a metabolism study be conducted on a root cr op and that the radi olabels include a radiolabel on the cyclopropyl ring.

860.1480 Meat/Milk/Poultry/Eggs For livestock, any feeding st udy conducted in the future for additional uses should determine cyclopropylcarboxylic acid (CPCA). [CPCA should be monitored in future ruminant feeding studies (for registration requests that include additional livestock feedstuffs) in lieu of a cyclopropyl labeled metabolism study (ROCKS memo, DP#377139, G. Kramer, 5/18/2010).]Page 9 of 94 2.2 Tolerance Considerations

2.2.1 Enforcement Analytical Method

Adequate multiresidue methods test data fo r cyflufenamid were submitted. Acceptable recoveries of cyflufenamid from a non-fatty ma trix (grape) were achieved under Protocol E. Acceptable recoveries from a fatty matrix (milk) were also achieved under Protocol F. HED

recommends that FDA multiresidue methods be used as the primary enforcement method. The

submitted data will be forwarded to the U.S. FDA for further evaluation.

Adequate residue analytical methodology is av ailable for tolerance enforcement. The LC/MS/MS method (Method 070276) was submitted for the determination of cyflufenamid residues in/on pome fruit, cu curbit vegetables, grapes, and strawberries. The proposed enforcement method (Method 070276), in combin ation with the FDA multiresidue method, meets the OPPTS Residue Chemistry Test Guid elines for acceptable tolerance enforcement methods (SOP Number ACB-019).

Method 070276 monitors one mass ion transition. Although not a requirement since there is an adequate multiresidue method, the Agency is requ esting that the petitione r revise the LC/MS/MS enforcement method (Method 070276) to provide two mass ion tr ansitions for analysis of cyflufenamid.

An enforcement method for livestock commod ities is not needed because tolerances for cyflufenamid residues of concer n in meat, milk, poultry, and eggs are not required to support the proposed uses based on the resu lts of the goat metabolism st udy and the calculated dietary

burden.

2.2.2 International Harmonization

There are currently no maximum residue limits (MRLs) established by Codex, Canada, or Mexico for cyflufenamid. An International Resi due Limit Status sheet is attached (Appendix D).

2.2.3 Recommended Tolerances

HED recommends for the following tolerances and residue definition:

Tolerances are established for residues of the fungicide cyflufenamid, including its metabolites and degradates, in or on the comm odities in the table below. Compliance with the tolerance levels specified belo w is to be determined by measuring only cyflufenamid, [ N(Z)]-N -[[(cyclopropylmethoxy)am ino][2,3-difluoro-6- (trifluoromethyl)phenyl]met hylene]benzeneacetamide.Page 10 of 94 Apple, wet pomace...............................................................0.10 ppm Berry, low growing, subgroup 13-07G, except cranberry ...0.20 ppm Fruit, pome, group 11...........................................................0.06 ppm Fruit, small vine climbing, subgroup 13-07F, except fuzzy kiwifruit ........................................................0.15 ppm Grape, raisin .........................................................................0.30 ppm Vegetable, cucurbit, group 9 ................................................0.07 ppm

Note: The petitioner voluntarily wit hdrew the proposed use on cranberry.

2.2.4 Revisions to Petitioned-For Tolerances

Table 2.2.4 compares proposed and recommended to lerance levels. The recommended tolerance levels, including increases for pome fruit and cucurbits, are based on the submitted magnitude of the residue data and the Agency’s tole rance spreadsheet as specified by the Guidance for Setting

Pesticide Tolerances Based on Field Trial Data SOP.

Table 2.2.4. Tolerance Summary for Cyflufenamid

Commodity Proposed Tolerance (ppm) Recommended Tolerance (ppm) Comments Correct Commodity Definition

Pome fruit crop group 11 0.05 0.06 Fruit, pome, group 11
Apple, wet pomace 0.10 0.10 Adequate apple processing data are

available.

Cucurbit crop group 9 0.05 0.07 Vegetable, cucurbit, group 9

Small fruit vine climbing subgroup except fuzzy kiwifruit; crop subgroup

13-07F 0.15 0.15 Fruit, small vine climbing, subgroup 13-07F, except fuzzy kiwifruit

Grape, raisin 0.30 0.30 Adequate grape processing data are

available.

Berry, low growing subgroup 13-07G 0.20 0.20 Berry, low growing, subgroup 13- 07G, except cranberry

2.3 Label Recommendations

2.3.1 Recommendations from Residue Reviews

A revised Section B/label is requested for Miltrex® 10 SC Fungicide (EPA Reg. No. 8033- pending) prohibiting the addition of a spray adjuva nt to the spray solutio n and including plantback intervals (PBIs) of 30 days for small grains and leafy vegetables an d 365 days for all other crops not on the label.

2.3.2 Recommendations for Worker Safety

There are no specifications for amount of suffici ent finished spray volumes for the ornamental uses. However, correspondence with the registrant indicated that the spray applications are madePage 11 of 94 in 20 - 200 gal/A. HED recommends that th is language is added to the label for CYFLUFENAMID 10 SC Fungicide (EPA Reg. No. 8033-pending) for clarification purposes.

2.3.3 Recommendations for Residential Safety

It should be noted that the proposed label states “Not fo r homeowner use”; however, HED assessed residential handler expos ure activities for the ornamental uses because there is a potential that homeowners could apply this product. HED recomm ends that the label language be revised to clarify the specific type of appli cator and setting (i.e., “a pplications made only by licensed pest control operators in co mmercial and reside ntial settings”).

3.0 Ingredient Profile

Cyflufenamid belongs to a new fungicide class, amidoximes, for which the pesticidal mode of action is yet to be defined. Cyflufenamid is active against all strains of powdery mildew, including those strains resistant to demethyla tion inhibitors and to strobulurin-type and benzimidazole-type fungicides. It has shown both protectant and cura tive activity on powdery

mildew.

Nippon Soda Company, Ltd. has proposed uses fo r cyflufenamid on cucurbit vegetables (Crop Group 9), pome fruit (Crop Group 11), small fr uit vine climbing except fuzzy kiwifruit (Subgroup 13-07F), low growing berry (Subgr oup 13-07G), except cranberry, and outdoor landscape and outdoor nursery ornamentals.

Cyflufenamid is formulated as suspension con centrates containing 0.85 lb ai/gal (10% ai). Cyflufenamid is applied as postemergence broa dcast foliar sprays using ground or aerial equipment. Application through a ny type of irrigation system is prohibited. A maximum of two applications can be made to agricultural cr ops and ornamentals for a maximum of 0.022 lb ai/A/growing season (cucurbits), 0.044 lb ai/A/growing season (o ther agricultural crops), and 0.044 lb ai/A/year (ornamentals). For agricultural crops, retreatment interv als (RTIs) are 7 or 14 days and preharvest intervals (P HIs) are 0, 3, or 14 days. Retr eatment intervals for ornamentals are 14-21 days.

3.1 Structure and Nomenclature

Table 3.1. Cyfluf enamid Nomenclature

. Chemical Structure

F

F N H N O O

CF 3Page 12 of 94 Table 3.1. Cyfluf enamid Nomenclature

. Empirical Formula C 20 H 17 F 5 N 2 O 2 Common Name Cyflufenamid

Company experimental name NF-149

IUPAC Name (Z)- N -[ α -(cyclopropylmethoxyimino)-2,3-difluoro-6-(trifluoromethyl)benzyl]-2- phenylacetamide

CAS Name [( N ( Z) ]- N -[[(cyclopropylmethoxy)amino][2,3-difluoro-6- (trifluoromethyl)phenyl] methylene]benzeneacetamide

CAS Registry Number 180409-60-3

End-use product (EP) MILTREX® 10 SC Fungicide (0.85 lb ai/gal suspension con centrate; EPA Reg. No. 8033-RNG)

Chemical Class Fungicide/amidoxime

Known Impurities of Concern None

3.2 Physical/Chemical Characteristics

Table 3.2. Physicochemical Properties of Cyfluf enamid (Technical Grade of the Active Ingredient).

Parameter Value Reference

Molecular weight 412.36 g/mole Not applicable

Melting point/range 61.5-62.5 oC Administrative Document for Cyflufenamid: Petition for Food Crop Tolerances for the Cucurbits Crop Group, the Pome Fruit Crop Group, Grapes [and Other Climbing Vine Small Fruit (except Fuzzy Kiwifruit)], and Strawberries (and Other Low Growing Berries), dated 11/28/08 pH 6.2 (1% w/w solution)

Density 1.347 g/cm 3 at 20 oC

Water solubility 0.80 mg/L at pH 6.3-6.8 and 10 oC 0.52 mg/L at pH 6.3-6.8 and 20 oC 0.047 mg/L at pH 6.3-6.8 and 30 oC 0.014 mg/L at pH 4 and 20 oC 0.12 mg/L at pH 10 and 20 oC

Solvent solubility (20°C to 25°C) Not available

Vapor pressure 3.54 x10 -5 Pa at 20 oC

Dissociation constant, pK

a 12.08 at 20 oC

Octanol/water partition coefficient, Log(P OW ) at 20 oC 4.68 at pH 4.05 4.70 at pH 6.75 4.55 at pH 9.95

UV/visible absorption spectrum Neutral – λ

max 207, 238 nm

Acidic – λ

max 207, 238, 361 nm

Basic – λ

max 220, 240 nm

3.3 Summary of Existing/Proposed Uses

There are no existing uses in the U.S. Cyflufen amid is registered in Japan, Korea, and several European Union (EU) countries.

Nippon Soda has proposed uses on agricultural crops (cucurbit vegetables, gr apes and other vine climbing small fruits, pome fru it, and strawberries and othe r low growing berries, except cranberry) and ornamentals as summarized in Tabl e 3.3 below. The formulation for agricultural crops is MILTREX ® 10 SC Fungicide (EPA Reg. No. 8033- pending), a 0.85 lb ai/gal suspension concentrate (SC) formulation. The formulation for ornamental uses is CYFLUFENAMID 10 SCPage 13 of 94 Fungicide (EPA Reg. No. 8033-pend ing), which is also a 0.85 lb ai/gal SC formulation.

The submitted label for Miltrex® 10 SC Fungicide (EPA File Symbol 8033-RNG) is adequate to

allow evaluation of the residue data relative to the proposed uses. Refer to Section 2.3 for recommended modifications to the proposed labels.

Table 3.3. Summary of Directions for Use of Cyflufenamid

Applic. Timing, Type,

and Equip. Formulation [EPA Reg. No.] Applic. Rate (lb

ai/A) Max. No. Applic. per

Growing

Season Max. Seasonal Applic. Rate

(lb ai/A) PHI (days) Use Directions and

Limitations 1

Cucurbit Vegetables: Pumpkin, Squash (summer and winter), Watermelon, and other Melons such as Chayote (fruit), Chinese waxgourd (Chinese preserving melon), Citron melon, Cucumber, Gherkin, Gourd (edible), Momordica spp., Muskmelon (hybrid and/or cultivars of Cucumis melo including true cantaloupe, cantaloupe, casaba, chreshaw melon,

golden pershaw melon, honeydew melon, honey balls, mango melon, Persian melon, pineapple melon, Santa Claus melon, and snake melon)

Postemergence Foliar spray Ground, aerial 0.85 lb/gal

SC [8033-RNG] 0.011 2 0.022 0 Applications should begin at first sign of disease development. Do not apply more than once every 7 days. Apply in minimum spray volumes of 5 GPA by air or 20 GPA by ground.

Grapes; Amur river grape; Gooseberry; Kiwifruit, hardy; Maypop; Schisandra berry; and cultivars, varieties, and/or hybrids of these

Postemergence Foliar spray Ground, aerial 0.85 lb/gal

SC [8033-RNG] 0.011-

0.022 2 0.044 3 Applications should begin at first sign of disease development. Do not apply more than once every 7 days at the low rate and 14 days at the high rate. Apply in minimum spray volumes of 5 GPA by air or 20 GPA by ground.

Pome Fruit: Apple, Pear, Crabapple, Loquat, Mayhaw, Quince, Oriental pear

Postemergence Foliar spray Ground, aerial 0.85 lb/gal

SC [8033-RNG] 0.022 2 0.044 14 Applications should begin at first sign of disease development. Do not apply more than once every 14 days. Apply in minimum spray volumes of 10 GPA by air or 50 GPA by ground.

Strawberries and other low growing berries except cranberry: Bearberry; Bilberry; Lowbush Blueberry; Cloudberry; Lingonberry; Muntries; Partridgeberry; and cultiv ars, varieties, and/or hybrids of these

Postemergence Foliar spray Ground, aerial 0.85 lb/gal

SC [8033-RNG] 0.011-

0.022 2 0.044 0 Applications should begin at first sign of disease development. Do not apply more than once every 14 days. Apply in minimum spray volumes of 10 GPA by air or 20 GPA by ground.

Outdoor Landscape and Outdoor Nursery Ornamentals 2Page 14 of 94 Table 3.3. Summary of Directions for Use of Cyflufenamid

Applic. Timing, Type,

and Equip. Formulation [EPA Reg. No.] Applic. Rate (lb

ai/A) Max. No. Applic. per

Growing

Season Max. Seasonal Applic. Rate

(lb ai/A) PHI (days) Use Directions and

Limitations 1

Postemergence Foliar spray Ground, aerial 0.85 lb/gal

SC [8033- pending] 0.011-

0.022 2 per year 0.044 per year NA Apply in sufficient finished spray volume appropriate for the size of the plant and amount of foliage. Use the higher rates and shorter interval under high

disease conditions.

HED assumed applications are made by groundboom, airblast, backpack, hose-end sprayer or low pressure hand wand.

The retreatment interval is 14- 21 days.

1 Restrictions: Application of Miltrex® 10 SC Fungicide through irrigation equipment is prohibited. A restricted-entry interval (REI) of 12 hours is proposed. For disease resistance management, the number of sprays of Miltrex® 10 SC Fungicide per crop is not to

exceed 50% of the total numb er of sprays for powdery mildew. The label recommends that the product be alternated with other mode of action fungicides. There are no rotational plant-back restrictions lis ted on the submitted draft label for Miltrex® 10 SC Fungicide.

2 The outdoor ornamental and nursery cr ops include abelia, acalypha, achillea, ageratum, amelanchier, amorpha, aster, azalea, barberry, begonia, bellflo wer, bittersweet, buckeye, buttonbush, California poppy, catalpa, flowering cherry, horse chestnut, cosmos, flowering crabapple, crepe-myrtle, dahlia, delphinium, dogwood, elm, euonymus, fleabane, gaillardia, gardenia, geranium, gerbera daisy, ornamental gourd, grape leaf ivy, hackberry, hawthorn, hibiscus, holly, hollyhock, honeysuckle, lilac, locust, maple, mock-orange, mountain laurel, oak, pansy, flowering pear, petunia, phlox, photinia, privet, rhododendron, rose, salvia, sedum, snapdragon, sycamore, sunflower, trumpet creeper, viburnum, walnut, willow and zinnia.

4.0 Hazard Characterization and Dose-Response Assessment

4.1 Toxicology Studies Available for Analysis

The toxicology database for the ne w active ingredient, cyflufenamid, is considered adequate for risk assessment in support of th e current registration action under FIFRA Section 3 for food use. Toxicity studies submitted in support of registra tion are detailed in the following section and a brief summary of the findings is discusse d in Section 4.3, Toxicological Effects.

Data from the following studies were used to evaluate the hazard pot ential of cyflufenamid:

• Acute: Acute oral toxicity of cyflufenam id, technical and its metabolite/degradate 149- ( E )-FB in rats; acute dermal, rat; acute eye inhalation, rat; acute eye irritation, rabbit; acute dermal irritation, rabbit; and dermal se nsitization, guinea pig. A waiver request for the acute neurotoxicity screen was submitted.

• Subchronic: 13-week oral, ra t; 13-week oral, mouse; 13-we ek oral, dog; 28-day dermal,Page 15 of 94 rat; and subchronic neurotoxicity, rat.

• Chronic: 52-week oral, dog; 108-week oral, mouse; 2-year oral/carcinogenicity, rat.

• Reproduction/developmental: Developmen tal toxicity, rabbit; developmental toxicity/one-generation reproduction, rat; and two-generation reproduction, rat.

• Other: 6 genotoxicity screens (including 1 isomer), 3 metabolism studies in rats, 2 immunotoxicity studies (mouse and rat) and 3 mechanistic toxicology studies.

4.2 Absorption, Distribution, Metab olism, & Elimination (ADME)

A difference in the excretion pa ttern between sexes was noted in the rat metabolism study. The rate of urinary excretion in male s was greater than twice that of females. Consequently, fecal excretion was less in males (66% administered dose) and higher in females (81%). Biliary excretion was greater in females than males indicating increased absorption in female rats. A substantial proportion of the biliary radioactivity was ultimately elim inated in the feces of rats treated with a single low dose of cyflufenamid, indicating substantia l enterohepatic recirculation.

There was no substantial difference in excreti on pattern between single and repeat doses of cyflufenamid but absorption was greater in rats treated with a single low dose (10 mg/kg) than a single high dose (200 mg/kg).

Overall tissue accumulation of ra dioactivity was low; less than 1% was present 168 hr postdosing. Concentrations of radioactivity were high est in the liver, kidney a nd erythrocytes and fat (of females treated with 200 mg/kg). Especially l ong half-lives (t1⁄2) were noted for uterus, ovary

and fat in females and red blood cells in both sexe s. The elimination half-life of plasma was 14-

34 hrs while it exceeded 70 hours in erythrocytes. This longer t1⁄2 may be explained by the affinity of fluorinated phenyl moie ties for membranes of red blood cells.

Following administration of e ither [fluorinated phenyl-U- 14 C] or [cyclopropyl-2,3- 14 C]NF-149, the major metabolite identified in urine was 149-F1 (5 -14% of the administered dose). (Refer to Appendix B for chemical names and structures). Treatment with β -glucuronidase or sulphatase showed that urinary metabolites were, for the most part, not conjugated. No unchanged parent was detected in urine. The major meta bolite in feces was 149-F-3-OH-4-OH-B (11-27% administered dose). Unchanged parent accounte d for ~5% of administered dose in animals treated with 10 mg/kg and increased as ab sorption became saturated at the higher and consecutive doses; unchanged cyflufenamid represen ted ~30% of the administered dose in rats given 10 mg/kg/day for 14 days. Biliary metabolite s were qualitatively si milar regardless of the dose and suggested extens ive hepatic metabolism in cluding glucuronidation.

The proposed metabolic pathways, based on th e observed excreted metabolites in rats administered radiolabeled cyflufenamid, ar e pictured in Figure 4.2. In one pathway, cyflufenamid is hydrolyzed to 149-F which is reduced to the amidine, 149-F1, and then subsequently deaminated to form 149-F6. In the other main pathway, 149-F-3-OH-4-OH-B was formed by di-hydroxylation of the non-fluorinated phenyl ring and subsequently converted to the methoxy derivative. Hydroxylation of the α position occurred as a minor pathway yielding 149-

F-3-OH-B and 149-F-4-OH-B. 149-F4 B was glucuronidated or cycl ized to the oxadiazole, 149-

F12.Page 16 of 94 Figure 4.2

4.2.1 Dermal Absorption

A non-guideline in-vitro study was submitted wh ich provides information on the dermal penetration of cyflufenamid (NF- 149) following a single applica tion to excised human skin of [ 14 C]NF-149 in a SC (suspension concentrate) fo rmulation at two dose levels. At 1000 μg/cm 2

, the total absorbed dose was only 1.14% of the applied dose (AD), al most entirely recovered in

the stratum corneum (1.11% AD). The absorption rate was 0.0118 μg/cm 2 /h. At 0.136 μg/cm 2

, the total absorbed dose was 13.71% AD, almost entirely isolated in the stratum corneum (12.93% AD) and the absorp tion rate was 0.00005 μg/cm 2 /h. Total recovery was 93% at each dose. In the absence of a dermal absorption study and an in-vitro study in excised rat skin, this study cannot be relied upon to provide a dermal ab sorption factor (DAF) for this risk assessment.

Since an acceptable/guideline dermal absorption study in the rat was not available, a dermal absorption factor was calculated as the ratio of the subchronic oral LOAEL, 117.4 mg/kg/day, to the dermal NOAEL, 1000 mg/kg/day, from th e 28-day dermal toxicity study (LOAEL not established); the upper bound estimate of dermal absorption, DAF, is 12%. Note that liver parameters were measured in the 28-day derm al toxicity study but hepatotoxicity was not observed. Liver is the primary organ of cyflufenamid toxicity.Page 17 of 94 4.3 Toxicological Effects

In the mammalian toxicology database, the liver is the primary target organ for cyflufenamid toxicity. Across species, durat ion and gender, changes in we ight, clinical chemistry and pathology indicate treatment-relate d perturbations in and advers e effects on liver function. Common, toxicologically si gnificant changes in clinical chemis try include: increased alkaline phosphatase activity; increased gamma glutamyl tr ansferase activity; incr eased cholesterol and triglycerides; increased alanine and aspartate tr ansferases; increased urea nitrogen and increased total protein. Increased liver weight is ofte n accompanied by periacinar hepatocytic hypertrophy and vacuolation. Fatty liver changes observed in the subchronic and chronic mouse studies may be evidence of cyclopropane car boxylic acid (CPCA) toxicity.

Thyroid effects due to treatment with cyflufen amid were seen only in the rat. Increased follicular cell hypertrophy (as well as increa sed organ weight) was noted in the 90-day subchronic feeding study. Neoplas tic thyroid follicular adenomas were noted in the 2-year carcinogenicity study. The submitter postulated that thyroid changes were considered to be secondary to a treatment-relate d increase in metabolic activity of the liver but sufficient mechanistic data to support th is hypothesis were not submitted.

Kidney effects related to treatm ent were observed in the subchr onic and chronic rat studies. Increased kidney weight was accompanied by tubular vacuolation and slight decreases in sodium

and chloride concentrations.

Treatment-related cardiotoxicity was noted in the rat and mouse feedi ng studies. Observed myocardial vacuolation and lip idosis can be attributed to decreased lipid metabolism; cyflufenamid caused an approximately 50% inhibition of carnitine palmitoyltransferase in both

rat and mouse heart microsomal fractions in a non-guideline mechanistic study. Carnitine palmitoyltransferase is involved in the transport of long chain fatty acids into the mitochondrial matrix for oxidation. Fatty acid oxidation is an important source of energy for ATP production

in the mitochondria.

Cyflufenamid-induced brain vacuolation was specif ic to the dog and not associated with any apparent clinical sign of neur otoxicity. Supplementary studies investigating this phenomenon determined that vacuolation was due to myelin edema affecting the white matter of the cerebrum and thalamus. Furthermore, this brain lesion wa s partially reversed after a 13-week recovery period (following 90-day exposure) and fully reversed after a 26-w eek recovery period. This effect was not observed in any other species. A subchronic neurotoxicit y study in rats showed no evidence of neurotoxicity.

Effects on reproductive organs and/or parameters were noted in severa l subchronic studies at doses greater than the respect ive LOAELs. Decreased uterus and cervix weights, adrenal cortical hypertrophy and reduced quality and quantity of spermato zoa were observed in dogs. Leydig cell hypertrophy was observe d in rats and mice. It is unclear what toxicological significance should be ascribed to these findings since they may be secondary to systemic toxicity or hepatic enzyme induction. Ma ting performance and fertility in the P/F 0 generation were both unaffected by treatment with cyflufen amid in the 2-generation reproductive toxicityPage 18 of 94 study in rats. Sex ratio, sexual maturation, estr ous cyclicity, sperm quantity and quality, mating performance and fertility, gestati on and viability indices in the F 1 generation were all unaffected by treatment.

Study summaries of the acute, subc hronic and chronic toxi city and genotoxicity for cyflufenamid are provided in Appendix A at the end of this document.

Cyflufenamid has low acute toxicity via the oral , dermal and inhalation routes of exposure. Though slightly irritating to the eye, cyflufenamid is not a skin irritant or sensitizer under the conditions of the Guinea Pig Maximization Test. The Acute Toxicity Profiles are tabulated in Appendix A2. It is pertinent to note that some environmental degradates/metabolites are apparently more acutely toxic than the pare nt compound. Compound 149-F 1, in particular, is classified in Toxicity Category II. However, si nce 149-F1 is a major urinary metabolite in rats administered cyflufenamid, its subacute – chro nic toxicity is accounted for in the mammalian toxicology database.

Cyflufenamid is classified as “likely to be car cinogenic to humans.” This was based on the presence of two tumor types in two species: thyroid follicular cell tumors in male rats and liver

tumors in male mice. There is no concern for mu tagenicity or clastogeni city. The unit risk, Q 1 *

, of cyflufenamid based upon male mouse liver co mbined adenoma and carcinoma tumor rates is

6.61 x 10 -3 (mg/kg/day) -1 in human equivalents. Two non-guideline studies were submitted to address thyroid lesions but we re not sufficient to support a carcinogenic mode of action.

4.4 Safety factor for Infants and Children (FQPA Safety Factor)

Based on the hazard and exposure data, the cyfluf enamid risk assessment team has recommended that the FQPA Safety Factor be reduced to 1X because there is an adequate toxicity database for cyflufenamid and exposure data are complete or are estimated based on data that reasonably account for potential exposures. There is no evidence of suscep tibility following in utero and/or postnatal exposure in the developmen tal toxicity studies in rats or rabbits, and in the 2-generation rat reproduction study. There are no residual uncertain ties concerning pre- a nd postnatal toxicity and no neurotoxicity concerns. The chroni c and cancer dietary (food and drinking water) exposure assessments are conservative. The di etary and residential assessments are based on reliable data and will not underestimate exposure/risk. Based on these data and conclusions, the FQPA Safety Factor can be reduced to 1X.

4.4.1 Completeness of the Toxicology Database

The toxicology database is complete with the ex ception of an acute ne urotoxicity study (ACN, OPPTS 870.6200a) which is a new data requirement under 40 CFR Part 158 for registration of a pesticide (food and non-food uses). EPA has ev aluated the available toxicity data for cyflufenamid and determined that an additional database uncertainty factor is not needed to account for potential neurotoxi city. The reasons follow:Page 19 of 94 • An acute neurotoxicity screening test will unlik ely yield a point of departure less than the chronic NOAEL of 4.4 mg/kg/day. At this level, there would be no risk concern based on acute dietary exposure.

• In an acute oral toxicity study, adverse effects were noted on the day of administration (limit dose) but not thereafter; clinical si gns of piloerection, hunched posture, unsteady gait, pallid extremities, increased sa livation, ungroomed appearance and abnormal respiration were observed in the majority of animals receiving 5000 mg/kg and generally resolved by Day 2 of the study.

• An acceptable, guideline subchronic neurotoxic ity study was submitted. Repeat exposure to doses up to approximately 500 mg/kg/day di d not elicit any neurotoxic effects as

assessed in the functional obs ervational battery, motor activ ity, neurohistopathology or brain morphometrics.

• Cyflufenamid is not an apparently neurot oxic chemical based on clinical toxicity assessments incorporated within the developm ental and chronic rat studies. In several short-term studies in rats (subacute and subchronic feeding, plaque-forming cell assay, one-generation pilot, developmental toxicity), no neurobehavioral si gns were observed at the highest doses tested. While the relevant and reversible effect of brain vacuolation was observed in the subchronic dog study at ap proximately 70 mg/kg/day, it is observed

in the absence of overt neur otoxicity and nowhere else in the toxicology database. In summary, while the results of the required acute neurotoxicity study may show an adverse effect following a single dose, ther e is no need to retain an UF DB , as any point of departure (POD) based on neurotoxic effects is likely to be at least 100 fold higher than the chronic RfD.

Based on the available toxicity da tabase and the Agency's current practices, the inhalation risk for cyflufenamid was assessed using an oral t oxicity study. The Agency sought expert advice and input on issues related to th is route to route extrapolation a pproach (i.e., the use of oral toxicity studies for inhalation risk assessment) from its Fede ral Insecticide, Fungicide, and

Rodenticide Act Scientific Advi sory Panel (SAP) in December 2009. The Agency received the SAP’s final report on March 2, 2010 ( http://www.epa.gov/scipoly/SA P/meetings/2009/120109meeting.html ). The Agency is in the process of evaluating the SAP report and may, as appropriate, re-examine and develop new policies and procedures for conducting inhalati on risk assessments, in cluding route to route extrapolation of toxicity data. If any new polic ies or procedures are developed, the Agency may

revisit the need for an inhalation toxicity study for cyflufenam id and/or re-examine the inhalation toxicity risk assessment.

4.4.2 Evidence of Neurotoxicity

Cyflufenamid is not an apparently neurotoxic chemical based on clinical observations of neurotoxicity during the conduct of developm ental or chronic studies. No untoward neurobehavioral signs were obser ved at doses approaching the lim it dose in any of the shortterm studies in rats or mice (subchronic oral, im munotoxicity, 28-day dermal) or chronic studies.

No neurotoxic effects were observed during the subchronic ne urotoxicity battery in which animals were dosed at systemically toxic dos e levels (572 mg/kg/day) . A requisite acute neurotoxicity screen has not been submitted for this new active ingredient; this requirement shall remain as a condition of registration. Myelin ed ema, diffuse vacuolation of white matter, wasPage 20 of 94 observed in dogs after 90 days of treatment with 76.2 mg/kg/day. This effect was not seen in the chronic dog study, largely in part, because the HDT was 17.3 mg/kg/ day. Brain vacuolation may be regarded as a reversible effect since this lesion was less severe after a 13-week recovery period and absent after a 26-week recovery period (both followi ng a 90-day treatment). Brain vacuolation may also be regarded as species-specific since there were no microscopic findings of the nervous system in either the rat or mouse. It is pertinent to note th at neurological function was not altered in dogs treated with 76.2 mg/kg/da y for 90 days. The endpoints selected for this risk assessment are protective of this effect seen in dogs.

A developmental neurotoxicity study is not required at this time.

4.4.3 Evidence of Sensitivity/Susceptibility in the Developing or Young Animal

Neither the rat nor rabbit devel opmental studies identified terat ogenic effects. There was no evidence of increased susceptibility in the submitted toxicology database.

There is no concern for increased susceptibility to offspring followi ng pre- and postnatal
exposure to rats or in utero exposure in rabbits. Maternal NOAELS are clearly defined and do not exceed offspring values and are based on genera l systemic toxicity. The marginally higher incidence of incompletely ossified epiphyses a nd metacarpals/phalanges seen in rabbits may be associated with low fetal weight and are indicat ive of delayed embryo-fetal development. The combined offspring effects of decreased body wei ght and incomplete ossification are believed to

be related to the observed maternal toxicity. Furthermore, the point s of departure selected for all exposure scenarios are lower than those dos es causing adverse effects in offspring.

Maternal treatment with cyflufenamid had no de velopmental effects in rats. A dose-dependent reduction in fetal body weight was ob served in rabbits treated with ≥ 60 mg/kg/day. A higher incidence of incompletely ossified epiphyses and metacarpals/phalanges was also observed at mid-dose and above. Both effects may be regarded as secondary to maternal toxicity (decreased body weight gain) rather than a direct effect upon the fetus by cyflufenamid. Cyflufenamid has no teratogenic potential up to the limit dose of 1000 mg/kg/day in rats and did not produce any developmental effects in rabbits at doses lower than the maternal LOAEL. Offspring toxicity was limited to reduced body weight and body weight ga in at doses equal to those eliciting similar effects in the parental generations in th e 2-generation reproduc tive toxicity study.

4.4.4 Residual Uncertainty in the Exposure Database

There are no residual uncertainti es in the exposure database. Since the dietary and non-dietary exposure estimates were based on several conser vative assumptions, HED does not believe that the exposure estimates are underestimated. Th e chronic dietary asse ssment conducted with DEEM-FCID was a partially refined analysis. The assessment utilized average field trial residues for all crops. In addition, default proc essing factors were assumed for all commodities except for apple juice and grape juice where empirical processing factors were used. 100% crop

treated was assumed for all commodities. The D EEM analysis also assumed that all drinking water will contain cyflufenamid at the highest EDWC levels modeled by EFED for ground water

or surface water. For these reasons it can be concluded that the DEEM-FCID analysis does notPage 21 of 94 underestimate risk from chronic exposure to cy flufenamid. Similarly, HED does not believe that the non-dietary occupational and re sidential exposures are underest imated because they are also based on conservative assumptions, including ma ximum application rates, labeled PPE, and standard values for unit exposures , acreage treated/amount handled.

4.5 Toxicity Endpoint and Point of Departure Selections

The details for selecting toxicity endpoints and points of depa rture are presented in Appendix

A3. An acute reference dose was not establis hed as there were no adverse effects in the toxicology database which could be attributed to a single dose. The chronic reference dose was based on liver, thyroid and kidney effects obs erved in the combined chronic toxicity study/carcinogenicity study in rats . No dermal hazard was identif ied. The toxicity endpoint and point of departure observed in the single st udy, Prenatal Developmental Toxicity Study in Rabbits (decreased body weight gain and food c onsumption), are related to all other exposure scenarios and are protective of them (Section 4.5.4). The cyflufenamid risk assessment team has recommended that the FQPA safety fact or be reduced to 1X (Section 4.4).

4.5.1 Dose-Response Assessment

An acute reference dose was not established. Ab sent the required acute neurotoxicity screen, no other effects observed in the toxicology databa se were attributable to a single dose.

For chronic dietary exposure, a chronic reference dose of 0.04 mg/kg/day was selected for assessment of all populations based on a NOAEL of 4.4.mg/kg/day with evidence of hepatotoxicity at 22 mg/kg/day in the combined ch ronic toxicity/carcinogeni city study in rats. This provided the most sensitive endpoint availa ble for dietary exposure; chronic dog and mouse exposure NOAELs were 4- and 15-fold greater, respectively.

For short-and intermediate-term residential and oc cupational exposures involving incidental oral and inhalation exposure, the NOAEL of 5 mg/kg/da y from the prenatal developmental study in rabbits (supplementary data) was selected. The LOAEL of 10 mg/kg/day was based on decreased body weight gain and food consumption. A reproductive study in rats was available for incidental oral exposures scenarios but ha d higher NOAELs (18 mg/kg/day). Therefore, the rabbit developmental study which was chosen for inhalation exposure (adults) was also used as an incidental oral exposure s cenario endpoint to be protectiv e of children. The available multigeneration study also better represented the intermediate exposure duration but, likewise, was superseded by the chosen short-term PoD (rabbit developmental study, NOAEL = 5 mg/kg/day) which is more protect ive of longer term exposures.

This single endpoint, NOAEL = 5 mg/kg/day ba sed on decreased body weight gain and food consumption at 10 mg/kg/day, is conservative but justified by the observed dose-response relationship. The body weights of treated maternal rabbits were not signif icantly different from

control animals at the end of the study. However, it is pertinent to note that the rabbits in the control group were significantly smaller than a ll treatment groups on Day 0. In this case, a decrease in body weight change is a stronger indicator of general sy stemic maternal toxicity than body weight. Notable body weight decrement was s een in the Day 6-18 interval and sustainedPage 22 of 94 throughout. Furthermore, overall body weight gain s adjusted for gravid uterine weights were decreased by 107-171% in the trea ted groups compared to controls , reflecting maternal toxicity.

A 28-day dermal toxicity study in the rat, in which neuropat hology and hepatotoxicity were assessed, was available and showed no toxicity up to the limit dose. Since there were no concerns for developmental or neurological to xicities, dermal endpoints were not selected.

Long-term dermal and inhalation exposures are not expected; therefor e, endpoints were not

selected.

4.5.2 Recommendation for Combining Routes of Exposures for Risk Assessment

Residential exposures (dermal, or al, and inhalation) can be combin ed if the toxicological effects

are the same. Effects of d ecreased body weight, body weight gain and food consumption observed in the rabbit prenatal developmental study were selected for short-term/intermediateterm incidental oral and short-term/intermedi ate-term inhalation exposure for risk assessment; therefore, these routes of exposure can be combin ed to estimate aggregate risk. Since no hazard was identified in the dermal toxicology study and there are no concerns for developmental neurotoxicity, the dermal route of exposure need not be included in aggregate risk estimates.

4.5.3 Classification of Carcinogenic Potential

Cyflufenamid is classified as “likely to be car cinogenic to humans.” This was based on the presence of two tumor types in two species: thyroid follicular cell tumors in male rats and liver

tumors in male mice. There is no concern for mu tagenicity or clastogeni city. The unit risk, Q 1 *
(mg/kg/day) -1 , of cyflufenamid based upon male mouse liver combined adenoma and carcinoma

tumor rates is 6.61 x 10 -3 in human equivalents.

4.5.4 Summary of Points of Departure and T oxicity Endpoints Used in Human Risk

Assessment

Table 4.5.4.1. Summary of Toxicological Doses a nd Endpoints for Cyflufenamid for Use in Dietary and Non-Occupational Hu man Health Risk Assessments

Exposure/

Scenario Point of Departure Uncertainty/FQPA Safety Factors RfD, PAD,

Level of

Concern for

Risk

Assessment Study and Toxicological Effects

Acute Dietary (All Populations) no hazard identified

Chronic Dietary (All Populations) NOAEL=

4.4 mg/kg/day UF A = 10x

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

0.044 mg/kg/day

cPAD = 0.044 mg/kg/day Combined Chronic Toxicity/ Carcinogenicity Study in Rats LOAEL = 22 mg/kg/day based on decreased body weight gain; increased thyroid/parathyroid weight, increased liver weight and centrilobular hepatocytic hypertrophy.Page 23 of 94 Table 4.5.4.1. Summary of Toxicological Doses a nd Endpoints for Cyflufenamid for Use in Dietary and Non-Occupational Hu man Health Risk Assessments

Exposure/

Scenario Point of Departure Uncertainty/FQPA Safety Factors RfD, PAD,

Level of

Concern for

Risk

Assessment Study and Toxicological Effects

Incidental Oral Short-Term (1- 30 days) and

IntermediateTerm (1-6 months) NOAEL= 5 mg/kg/day UF A = 10x

UF H = 10x FQPA SF= 1x Residential

LOC for MOE

= 100 Prenatal Developmental Study in Rabbits Maternal LOAEL = 10 mg/kg/day based on decreased body weight, body weight gain and food consumption.

Dermal ShortTerm (1-30 days) and

IntermediateTerm (1-6 months) No adverse effects were observed in the dermal toxicity study and there are no concerns for developmental or neurolog ical toxicities; therefore, no hazards are expected from these exposure

scenarios.

Inhalation Short-Term (1- 30 days) and

IntermediateTerm (1-6 months) NOAEL = 5 mg/kg/day UF A = 10x

UF H = 10x FQPA SF= 1x Residential

LOC for MOE

= 100 Prenatal Developmental Study in Rabbits Maternal LOAEL = 10 mg/kg/day based on decreased body weight, body weight gain and food consumption.

Cancer (oral, dermal, inhalation) Classification: Likely to be carcinogenic to humans. Quantification of cancer risk was recommended. The Q 1 * value is 6.61 x 10 -3 (mg/kg/day) -1

.

Point of Departure (POD) = A data point or an estimated poin t that is derived from observed dose-response data and used to mark the beginning of extrapolation to determine risk associated with lower en vironmentally relevant human exposures. NOAEL = no observed adverse effect level. LOAEL = lowest observed adverse effect level. UF = uncertainty factor. UF A = extrapolation from animal to human (interspecies). UF H = potential variation in sensitivity among members of the human population (intra species). FQPA SF = FQPA Safety Factor. PAD = population adjusted dose (a = acute, c = chronic). RfD = reference dose. MOE = margin of exposure. LOC = level

of concern.

Table 4.5.4.2. Summary of Toxicological Doses a nd Endpoints for Cyflufenamid for Use in Occupational Human Health Risk Assessments

Exposure/

Scenario Point of Departure Uncertainty

Factors Level of Concern

for Risk

Assessment Study and Toxicological Effects

Dermal ShortTerm (1-30 days) and

IntermediateTerm (1-6 months) No adverse effects were observed in the dermal toxicity study and there are no concerns for developmental or neurolog ical toxicities; therefore, no hazards are expected from these exposure

scenarios.

Inhalation Short-Term (1- 30 days) and

IntermediateTerm (1-6 months) NOAEL = 5 mg/kg/day UF A = 10x

UF H = 10x Occupational LOC

for MOE = 100 Prenatal Developmental Study in Rabbits Maternal LOAEL = 10 mg/kg/day based on decreased body weight, body weight gain and food consumption.Page 24 of 94 Table 4.5.4.2. Summary of Toxicological Doses a nd Endpoints for Cyflufenamid for Use in Occupational Human Health Risk Assessments

Exposure/

Scenario Point of Departure Uncertainty

Factors Level of Concern

for Risk

Assessment Study and Toxicological Effects

Cancer (oral, dermal, inhalation) Classification: Likely to be carcinogenic to humans. Quantification of cancer risk was recommended. The Q 1 * value is 6.61 x 10 -3 (mg/kg/day) -1

.

Point of Departure (POD) = A data point or an estimated poin t that is derived from observed dose-response data and used to mark the beginning of extrapolation to determine risk associated with lower en vironmentally relevant human exposures. NOAEL = no observed adverse effect level. LOAEL = lowest observed adverse effect level. UF = uncertainty factor. UF A = extrapolation from animal to human (interspecies). UF H = potential variation in sensitivity among members of the human population (intra species). FQPA SF = FQPA Safety Factor. PAD = population adjusted dose (a = acute, c = chronic). MOE = margin of exposure. LOC = level of concern.

HED’s LOC for the MOE is defined by the uncertainty factors that are applie d to the assessment. HED applies a 10X factor to acc ount for interspecies extrapolation and a 10X factor to account for intraspecies sensitivity. Therefore, the total uncertainty factor that has been applied to the

non-cancer risk assessment is 100 for residential and occupational exposure. Residential and occupational risks resulting in MOEs greater than or equal to 100 (HED’s LOC) provide a health protective margin of exposure.

4.5.5 Endocrine Disruption

As required under FFDCA section 408(p), EP A has developed the Endocrine Disruptor Screening Program (EDSP) to determine whether certain substances (inc luding pesticide active and other ingredients) may have an effect in humans or wildlife si milar to an effect produced by a “naturally occurring estrogen, or other such endocrine effects as the Administrator may designate.” The EDSP employs a two-tiered ap proach to making the statutorily required

determinations. Tier 1 consists of a battery of 11 screening assays to identify the potential of a chemical substance to interact with the estrog en, androgen, or thyroid (E, A, or T) hormonal systems. Chemicals that go th rough Tier 1 screening and are found to have the potential to interact with E, A, or T hormonal systems will proceed to the next stag e of the EDSP where EPA will determine which, if any, of th e Tier 2 tests are necessary base d on the available data. Tier 2 testing is designed to identify a ny adverse endocrine related eff ects caused by the substance, and establish a dose-response relationship between the dose and the E, A, or T effect.

Between October 2009 and February 2010, EPA is issu ing test orders/data call-ins for the first group of 67 chemicals, which contains 58 pesticid e active ingredients and 9 inert ingredients.

This list of chemicals was selected based on th e potential for human exposure through pathways such as food and water, residen tial activity, and certain post-applic ation agricultural scenarios.

This list should not be construed as a lis t of known or likely e ndocrine disruptors.

Cyflufenamid is not among the group of 58 pesticid e active ingredients on th e initial list to be

screened under the EDSP. Unde r FFDCA Sec. 408(p), the Agency must screen all pesticide chemicals. Accordingly, EPA anticipates issuin g future EDSP test orders/data call-ins for all pesticide active ingredients.Page 25 of 94 For further information on the status of the ED SP, the policies and procedures, the list of 67 chemicals, the test guidelines and the Tier 1 screening battery, pleas e visit our website: http://www.epa.gov/endo/.

5.0 Dietary Exposure and Risk Assessment

5.1 Metabolite/Degradate Residue Profile

5.1.1 Summary of Plant and Animal Metabolism Studies

PRIMARY CROPS: Metabolism studies were c onducted in apples, cucumbers, and wheat uniformly labeled in the fluorin ated phenyl ring (apple, cucumber , and wheat) or on the 2 and 3 carbons of the cyclopropane ring (w heat). Positions of the radi olabeling are indicated below.

F

F N H N O O

CF 3 * F

F N H N O O

CF 3 * *

Figure 5.1. 14 C Labels in Metabolism Studies

In primary crops, unchanged cyflufenamid was the major residue identified/confirmed and

accounted for the bulk of total radioactive resi dues in various matrices including: apple fruit (66% of the total radioactive re sidues [TRR]), cucumber fruit (55-96% TRR), cucumber leaves (42-98% TRR), wheat forage (89-104% TRR) , and wheat straw (33-65% TRR). Parent accounted for 16.9% TRR in apple leaves and 2-7% TRR in wheat grain at mature harvest. Residues of cyflufenamid appeared to be highest in samples colle cted at early sampling intervals and were observed to decrease at longer interval s. Several minor struct urally related metabolites (<9% TRR) were detected, including the E -isomer of the parent compound. (The chemical names and structures of these compounds are depi cted in Appendix B.) The apple and cucumber

studies have shown that the bulk of the T RR was mainly surface residues which could be removed by rinsing. In addition, the cucumber st udy demonstrated that radioactive residues are not readily translocated from the soil into the aerial portion of the cucumber plant.

ROTATIONAL CROPS: A confined rotati onal crop study was conducted on wheat (small grain), lettuce (leafy vegetable) and carrot (root vegetable) with [phenyl-U- 14 C]cyflufenamid. Although TRR accumulated at >0.01 ppm in some rotated crop matrices following treatment at 1x the maximum proposed seasonal rate for rotata ble crops, the analytical work performed did not identify the parent cyflufenamid or any of its putative and structurally related metabolites. The only residue component identified was Meta bolite 149-F6 which was detected in maturePage 26 of 94 carrot foliage from the 120-day PBI, accounting for 32.5% TRR (0.013 ppm). Carrot foliage was not planted at a longer PBI.

RUMINANTS: A metabolism study was conducte d on lactating goats with cyflufenamid uniformly labeled on the fluorinated phenyl ri ng. The metabolism in lactating goats is characterized by the oxidation of the parent compound at the 3- and 4-positions of the unsubstituted phenyl ring to form Metabolite s 149-F-3-OH-B, 149-F- 4-OH-B and 149-F-3-OH4-OH-B; and cleavage of the amide bond forming 149- F. Further cleavage of the oxime ether bond forms 149-F1, and subsequent oxidation yiel ds 149-F6. The results at the high-dose feeding level of 13.3 ppm in goats showed that cy flufenamid was the major residue identified in fat accounting for 79.8% TRR. The parent was identi fied at lower levels in muscle (21.5% TRR) and milk (6.5% TRR). The parent was not detect ed in kidney or liver confirming the results from the low-dosed goat. The primary residue id entified in milk and tissues other than fat was 149-F1 accounting for 61.6% TRR in milk, 45.5% TRR in muscle, 55.5% TRR in kidney, and

29.6% TRR in liver. Metabolite 149 -F6 was also identified as a significant resi due in muscle (29.9% TRR) and kidney (13.9% T RR), and as a minor residue in liver (8.6% TRR) and milk (4.3% TRR). Metabolite 149-F was identified as a minor residue in fat (1.4% TRR) and liver (7.4% TRR). Hydroxylated meta bolites 149-F-4- OH-B and 149-F- α -OH-B were identified only in fat as minor residues (each ≤ 1.6% TRR). The isomer of cyflufenamid, 149-( E )-FB, was also only identified in fat, accounting for 2.5% TRR.

POULTRY: There are no poultry feedstuffs asso ciated with the proposed uses on pome fruit, cucurbit vegetables, vine climbing small fruits , and low growing berries. Therefore, data requirements for poultry metabolism are not relevant to this tolerance petition.

RAT: The metabolism of [fluorinated phenyl-U- 14 C]NF-149 in the rat pr oceeded via two main pathways. In one pathway, NF-149 was hydrolyzed to 149-F and then reduced to the amidine

149-F1. The 149-F1 was then deaminated to fo rm 149-F6. In the other main pathway, 149-F-3- OH-4-OH-B was formed by di-hydroxylation of the non-fluorinated phenyl ring and was subsequently converted to the methoxy deri vative. The intermed iate mono-hydroxylated

metabolites 149-F-3-OH-B and 149- F-4-OH-B were also presen t. Hydroxylation at the α position occurred as a minor pathway. A minor pathway proceeded via cleavage of the cyclopropylmethoxy moiety to form 149-F4B. The 149-F4B was then converted to a glucuronide conjugate. 149-F4B wa s also cyclized to the oxad iazole 149-F12. Conversion of NF-149 to the isomer 149-( E )-FB may represent a minor pathway. In the [cyclopropyl-2,3-

14 C]NF-149 groups, one or two hydroxylations of NF-149 took place, and these reactions sometimes proceeded to the formation of a 2-(cyclopropylcarbonylamino)acetic acid.

5.1.2 Summary of Environmental Degradation

Cyflufenamid is not expected to be mobile a nd it does not break down by abiotic processes, such as hydrolysis and photolysis. In the environment, its major route of dissipation is likely through

aerobic soil metabolism or runoff of soil to which cyflufenamid is absorbed. Aerobic metabolism studies indicate that cyflufenamid is moderately persistent . No degradation is

observed under anaerobic conditions. The half-liv es of cyflufenamid range from one week toPage 27 of 94 several months in various aerobic soil and a quatic metabolism studies. In one aerobic soil metabolism study cyflufenamid is shown to persist for more than a year.

Four degradation products of cyflufenamid ar e known, three major and one minor. The three major products are 149-F [ N -cyclopropylmethoxy-2,3-difluoro-6-t rifluoro-methylbenzamidine], 149-F1 [2,3-difluoro-6-trifluorom ethylbenzamidine], and 149-F11 [( Z )- N -( α - cyclopropyl methoxyimino-2,3-difluoro-6-trifluoromethylbe nzyl)carbamoylacetic acid]. The minor degradation product identified is 149-F6 [2,3-difluoro-6-triflu oromethylbenzamide]. These degradation products are mobile to very mobile compounds and have short to very long halflives.

It is believed that cyflufenamid may reach surf ace waters through spray dr ift and penetration of the canopy during application and foliar washoff to the soil followed by runoff of cyflufenamid absorbed soil. Field dissipation studies show th at cyflufenamid may pers ist in the environment for more than a year, increas ing the likelihood soil adsorbed cyflufenamid will contaminate surface water. In contrast, if cyflufenamid degr ades rapidly to more soluble compounds that are mobile or highly mobile as some of the aerobic soil metabolism data indicat e, it is possible that such degradates will contaminate surface wate r, as well as leach into groundwater.

5.1.3 Comparison of Metabolic Pathways

In primary crops, unchanged cyflufenamid was the major residue identified in various matrices.

In rotational crops, parent cyflufenamid wa s not identified. The only residue component identified was Metabolite 149-F6, which was detected in mature carrot foliage.

In drinking water, the residues of concern for risk assessment were parent, 149-F1, 149-F6, 149- F11, and 149-F.

In ruminants, parent was the major residue in fa t. The primary residue identified in milk and tissues other than fat was 149-F1, formed from 1 49-F. Metabolite 149-F6 was also identified as a significant residue in muscle and kidney, and as a minor resi due in liver and milk. The

residues of concern in ruminants for risk as sessment are cyflufenamid (Z-isomer), 149-F1, 149- F6, and cyclopropane carboxylic acid (c yclopropylcarboxylic acid; CPCA).

In the rat, the metabolism proceeded via two ma in pathways. In one pathway, parent (NF-149) was hydrolyzed to 149-F and then reduced to the amidine 149-F1. The 149-F1 was thendeaminated to form 149-F6. In the other main pathway, 149-F-3-OH-4-OH-B was formed by dihydroxylation of the non-fluorinated phenyl ring and was subsequent ly converted to the methoxy derivative. The intermediate mono-hydroxylat ed metabolites 149-F-3- OH-B and 149-F-4-OH-B were also present.Page 28 of 94 5.1.4 Residues of Concern Summary and Rationale

Table 5.1.4.1. Compounds to be Included in the Cyflufenamid Risk Assessment and Tolerance Expression. 1

Matrix Residues included in Risk

Assessment Residues included in Tolerance Expression for Compliance Monitoring

Plants Primary Crop* Cyflufenamid** Cyflufenamid

Rotational Crop Cyflufenamid Not required
Livestock Ruminant Cyflufenamid, 149-F1, 149-F6,

CPCA*** Not required

Poultry Not applicable Not applicable
Drinking Water Cyflufenamid, 149-F1, 149-F6, 149- F11, 149-F Not applicable

1 Memo of Residues of Concern Knowledge-based Subcommittee [ROCKS], DP#377139, G.F. Kramer, Ph.D.,

5/18/2010 Cyflufenamid = [ N(Z) ] -N- [[(cyclopropylmethoxy)amino][2,3-difluoro-6- (trifluoromethyl)phenyl]me thylene]benzeneacetamide.

149-F1 = 2,3-difluoro-6-trifluoromethylbenzamidine. 149-F6 = 2,3-difluoro-6-trifluoromethylbenzamide.

149-F11 = (Z)- N -( α -cyclopropylmethoxyimino-2,3- difluoro-6-trifluor omethylbenzyl)carbamoylacetic acid.

149-F = N -(cyclopropyl-methoxy-2,3-difluor o-6-trifluoromethylbenzamidine. CPCA = cyclopropane carboxylic acid. * for proposed crops only, additional data may be required for non-fruiting crops. ** not including the E -isomer. *** tentative, these residues should be in cluded in the feeding study (if required).

Rationale:

Plants

Primary Crops : The nature of the residue in plants is adequately understood for the presently proposed crops based on metabolism studies on apples, cucumbers, and wheat, pending submission of additional data and information for the submitted apple metabolism study. To upgrade the apple study, details of the in-life and analytical phases of the study along with quantitative raw data and sample storage information must be submitted.

In primary crops, cyflufenamid is the residue of concern for the tolerance expression and for risk assessment since unchanged cyflufenamid was the major residue. For the primary crops, cyflufenamid parent can serve as an adequate m easure of misuse based on its percentage of the

TRR in the submitted metabolism studies. Fo r dietary risk assessment purposes, ROCKS

concluded that no metabolites need to be included due to their low levels relative to the parent compound. The E -isomer of cyflufenamid can also be ex cluded as a residue of concern based on its low percentage of the TRR and since the an alytical enforcement method does not determine

the E -isomer.

HED does not have studies on three dissimilar crop s since both the apple and cucurbit can be considered to be fruits. If additional crops are proposed in the future (which are not similar toPage 29 of 94 apples, cucurbits, and wheat), a dditional metabolism data would be required; in that case, HED

recommends that a metabolism st udy be conducted on a root crop.

In the plant metabolism studies, although only one study (on wheat) included a radiolabel on the cyclopropyl ring, there was not a lot of cleavage of that ring so the submitted studies are adequate for the proposed crops. If a metabolism study is conducted in the future, the radiolabels

should include a radiolabel on the cyclopropyl ring.

Rotational Crops : The nature of the residue in rotati onal crops is adequately understood based on an acceptable confined rotati onal crop study conducted on wheat (small grain), lettuce (leafy vegetable) and carrot (root vegetable) with [phenyl-U- 14 C]cyflufenamid. Although TRR accumulated at >0.01 ppm in some rotated crop ma trices following treatment at 1x the maximum proposed seasonal rate for rotatable crops, the an alytical work performe d did not identify the parent cyflufenamid or any of its putative and st ructurally related metabolites. The only residue component identified was Metaboli te 149-F6 which was detected in mature carrot foliage from the 120-day PBI, accounting for 32.5% TRR (0.013 pp m). Carrot foliage was not planted at a longer PBI, so the rotational crop re striction on root crops needs to be set at the highest interval (365 days). Tolerances for inadvertent residue s of cyflufenamid in rotational crops are not

needed. The residue of concern for risk assessment is parent. Based on the rotational crop data, plant-back intervals of 30 days for small grains and leafy vegetables an d 365 days for all other crops which are not on the labe l are needed for the Miltrex® 10 SC Fungicide label.

Livestock

Ruminants: The nature of the residue in rumina nts is adequately understood based on an acceptable study conducted on lactating goats with cyflufenamid uniformly labeled in the fluorinated phenyl ring. In rumina nts (lactating goats), cyflufenam id was a major residue in fat and muscle; 149-F1 was a major residue in mil k, muscle, kidney, and liver; and 149-F6 was a major residue in muscle and kidney. Structur e-activity relationship (S AR) analysis (DEREK) revealed the same alert for the parent compound and all of these metabolites, polyhalogenated benzene- alpha-2-mu-Globulin ne phropathy (Table 5.1.4.2). However, this mode of action is not

relevant to humans since male ra ts are unique in their accumulation of this low molecular weight protein in the kidney.

Acute toxicity studies demonstr ated that 149-F1 was 10X more acutely toxic than the parent compound. However, since 149-F1 is also a ma jor metabolite in rats , sub-chronic/chronic toxicity studies with the parent are considered adequate to ac count for the toxicity of this metabolite. Due to structural similarities to the pa rent and in the absence of data to the contrary,

metabolite 149-F6 is also of concern.

Although not observed in the ruminant metabolism st udy (possibly due to the lack of a radiolabel in the cyclopropane ring), ther e is concern for the potential fo rmation of cyclopropane carboxylic acid (CPCA). The concern for CPCA is for a se vere form of liver toxicity, specifically

microvesicular steatosis. The concern was iden tified in a ROCKS review of another pesticide (aminocyclopyrachlor). The chemical s that cause this toxicity are typically short chain, branched carboxylic acids. The alert is in DEREK and CPCA seems to be in the training set of chemicalsPage 30 of 94 that support the alert, so there is no question that it falls within the domain of applicability. The formation of metabolites bearing the cycl opropanecarboxyl group in livestock commodities could be elucidated by analysis of these com pounds in a livestock f eeding study (see below).

A determination of the residues of concern in ruminants for the tolerance expression is not required at this time. A review of the livesto ck metabolism study and a livestock dietary burden

calculation indicate that a cattle feeding study and tolerances fo r livestock commodities are not

needed.

For risk assessment, the residues of c oncern in ruminants are cyflufenamid ( Z -isomer), 2,3- difluoro-6-trifluoromethylbenzamidine (149-F1), 2,3-difluoro-6-trifluor omethylbenzamide (149- F6) , and cyclopropane carboxylic acid (cyclopr opylcarboxylic acid; CPCA). CPCA should be monitored in future ruminant feeding studies (f or registration requests that include additional livestock feedstuffs) in lieu of a cyclopropyl labeled metabolism study.

Poultry and Swine: There are no poultry or swine feed items associated with the proposed uses.

Table 5.1.4.2. DEREK Alerts for Major Biodegradates. Compound Alerts Cyflufenamid 264 Polyhalogenated benzene. alpha-2-mu-Globulin nephropathy.

Number of matches = 1. 149-F1

149-F6

149-F11

149-F

CPCA 253 Short chain alkyl carboxylic acid. Irritation (of the gastrointestinal tract). Number of matches = 1. 546 Short chain fatty acid. Hepatotoxicity. Number of matches = 1.

Drinking Water: Three degradation products (149-F, 149-F1, and 149-F11) accumulated to >10% in a number of the submitted laboratory studies. Another degr adation product, 149-F6, had concentrations approaching 10% in some st udies. In addition, some study durations were not long enough to see clear formation and decline patterns for 149-F6. 149-F6 is thus likely to be a major degradate if the study durations were increased. As discussed above, degradation products 149-F1 and 149-F6 are of concern for risk assessment. Due to structural similarities to the parent and in the absence of data to th e contrary, degradation products 149-F11 and 149-F

are also of concern.

Recommended Tolerance Expression:
The ROCKS recommends the fo llowing language for the tolerance expression for plant

commodities:

Tolerances are established for resi dues of the fungicide cyflufenamid, including its metabolites and degradates, in or on the commodities listed below. Compliance with the tolerance levels specified below is to bePage 31 of 94 determined by measuring only cyflufenamid, [ N(Z) ] -N- [[(cyclopropylmethoxy)am ino][2,3-difluoro-6- (trifluoromethyl)phenyl]met hylene]benzeneacetamide.

5.2 Food Residue Profile

Crop Field Trials

The submitted magnitude of the residue data, summarized in Table 5.2.1, are adequate for risk assessment and tolerance assessment. For all crops , the number and locations of field trials are adequate. Four foliar broadcast applications of a 10% SC formulation of cyflufenamid were made to each crop. No adjuvants were used. Th e Agency’s tolerance spr eadsheet, as specified by the Guidance for Setting Pesticide Tolerances Based on Field Trial Data SOP, was used to determine appropriate tolerance levels. Based on the available data, all proposed tolerances are adequate, except for pome fruit and and cucurbit vegetables, for which tolerances of 0.06 and 0.07 ppm, respectively, are recommended.

Table 5.2. 1. Summary of Residues from the Crop Field Trials with Cyflufenamid.
Crop matrix Total Applic.

Rate (lb ai/A) PHI (days) Residue Levels (ppm)

n Min. Max. HAFT 1 Median Mean Std.

Dev.

Cucurbit Vegetables (proposed use = 0.022 lb ai/A total application rate, 0-day PHI)

Cucumber 0.044-0.046 0 12 <0.01 0.025 0.024 0.012 0.014 0.005

Cantaloupe 0.044-0.045 0 12 <0.01 0.032 0.025 0.015 0.016 0.007

Summer squash 0.044-0.046 0 10 <0.0 1 0.037 0.036 0.020 0.021 0.010
Pome Fruit (proposed use = 0.044 lb ai/A total application rate, 14-day PHI)

Apple fruit 0.088-0.092 13-15 24 <0.01 0.033 0.026 0.011 0.014 0.006

Pear fruit 0.088-0.090 14-15 12 0.014 0.044 0.037 0.022 0.024 0.009

Grapes and Small Vine Climbing Fruits (proposed us e = 0.044 lb ai/A total application rate, 3-day PHI)

Grape fruit 0.088-0.091 3 24 0.0125 0.0835 0.0750 0.0356 0.0370 0.0181
Strawberries and Other Low Growing Berries (proposed use = 0.044 lb ai/A to tal application rate, 0-day PHI)

Strawberry fruit 0.088-0.091 0 16 0. 015 0.134 0.118 0.072 0.072 0.032

1 HAFT = Highest average field trial result.

Processing Studies

The submitted apple and grape processing studies, summarized in Table 5.2.2, are acceptable. Residues of cyflufenamid concentrated in wet apple pomace (2.9x) but not in apple juice (0.3x) following processing of the RAC (whole apples) with quantifiable resi dues. Residues also concentrated in raisin (3.6x) but not in grape juice (0.2x) following processing of the RAC (grapes) with quantifiable residues. Following c onsiderations of the observed processing factors and highest average field trial (HAFT) residues, the proposed tolerances of 0.10 ppm for wet apple pomace and 0.30 ppm for raisin are adequate. No tolerances are required for apple juice and grape juice.Page 32 of 94 Table 5.2.2. Residue Data from Apple Processing Study with Cyflufenamid.

RAC Processed Commodity Total Rate (lb ai/A) PHI (days) Residues (ppm) [Average] Processing

Factor

Apple Fruit (RAC) 0.271 13 0.0305 --

Juice <0.01, <0.01 [<0.01] 0.3x

Wet pomace 0.087, 0.089 [0.088] 2.9x

Grape Fruit 0.271 3 0.106 --

Juice 0.029, 0.023 [0.026] 0.2x

Raisins 0.385, 0.379 [0.382] 3.6x

Storage Stability

Adequate storage stability data for cyflufenamid are available to validate the storage conditions and durations of storage of samples from the crop field trial and processing studies. Corrections

to residue values from the residue trials to account for dissipation during storage are not

necessary.

Meat, Milk, Poultry, and Eggs

The only livestock feed item associated with the proposed uses is wet apple pomace, which can be fed to dairy cattle; there are no beef cattle, hog, or poultry feedstuffs associated with the proposed uses ( Table 1 Feedstuffs , June 2008). A maximum reasona bly balanced dietary burden of 0.025 ppm for cyflufenamid residues in dairy cattle was calculated based on residues in wet apple pomace.

The results of the goat metabolis m study and the calculated dietary burden indicate that there is no reasonable expectation of finite cyflufenamid residues of concern in meat, milk, poultry, and eggs [40 CFR 180.6(a)(3)] as a re sult of the proposed uses; therefore, a ca ttle feeding study and tolerances on livestock commodities are not requi red. HED will re-examine this decision in the event that additional livestock feedstuffs are propos ed for registration in the future. Any feeding study conducted in the future for additional uses should determine cyclopropylcarboxylic acid (CPCA).Page 33 of 94 Rotational Crops

No tolerances are required for rotational crops. The confined rotational crop study indicated that no residues of concern are expected to occur in rotational crops with appropriate plant-back intervals. The proposed Section B/label must be revised to include plan t-back intervals of 30 days for leafy vegetables and small grains a nd 365 days for all othe r crops not on the label including root and t uber vegetables.

5.3 Water Residue Profile

Estimated surface water and groundwater c oncentrations are shown in Table 5.3.1. The dietary exposure assessment used the valu e of 4.68 ppb for chronic (non-cancer and cancer) assessments. The drinking water assessment is based on Tier I groundwater model simulations for all currently proposed cyflufenamid uses. (Cranberry is excluded since the petitioner voluntarily withdrew th e use on cranberry).

The drinking water assessment takes into account cyflufenamid, its isomer 149-( E )-FB [( E)

- N

- ( α -cyclopropylmethoxyimino-2,3-di fluoro-6-trifluoromethylbenz yl)-2-phenylacetamide], and four known degradation products of equal toxicological concern, 149-F [ N-cyclopropylmethoxy2,3-difluoro-6-trifluoro- methylbenzamidine], 149-F1 [2,3-difluoro-6- trifluoromethylbenzamidine], 149-F11 [( Z )- N -( α -cyclopropylmethoxyimino-2,3-difluoro-6- trifluoromethylbenzyl)carbamoylacetic acid], and 149-F6 [2,3-difluor o-6-trifluoromethylbenzamide]. These compounds are referred to colle ctively as total toxic cyflufenamid residues.

Table 5.3.1 Summary of Estimated Surface Water and Groundwater Concentrations for Cyflufenamid. c

Scenario Surface Water Conc., ppb a Groundwater Conc., ppb b

Acute 1.14 4.68

Chronic (non-cancer) 0.03 Not available

Chronic (cancer) 0.01 Not available

a From the Tier II PRZM-EXAMS - Index Reservoir model. Input parameters are based on 2 applications at 0.022 lbs ai/A/application to North Carolina apples using a percent cropped area (PCA) correction of 87%.

b From theTier I SCI-GROW (Screening Concentration in Groundwater) model assuming a maximum use rate of two applications at 0.022 lb ai/A/application, a K

oc of 10.5 mL/g, and an aerobic soil metabolism half-life of 314 days.

c DP#362954

, Rochelle F. H. Bohaty, Ph.D., 5/3/2010.Page 34 of 94 5.4 Dietary Risk Assessment

5.4.1 Description of Residue Data Used in Dietary Assessment

The chronic dietary analysis used for both non- cancer and cancer risk assessment assumed average field trial residues for all proposed cr ops. Empirical processing factors for apple juice and grape juice were used. (R efinements, made to upper bound exposure estimates specifically for use in the cancer assessment, were applied to both chronic and cancer assessments.) A separate tolerance was set for grape, raisin; therefore, the processing factor for this commodity was set at 1. For all other processed comm odities, DEEMTM version 7.81 default processing

factors were assumed.

Table 5.4.1 summarizes the cyflufenamid residue le vels and processing f actors for the proposed crops used in the chronic dietary assessments.

Table 5.4.1. Residue Data and Processing Factors used for Chronic Dietary Risk Assessment. Commodity Residues (Average; ppm) Processing Factors

Fruit, pome, group 11 Apple-0.014

Pear-0.024 Apple, juice = 0.3

Vegetable, cucurbit, group 9 Cucumber-0.014 Cantaloupe-0.016Squash, Summer0.022

Fruit, small vine climbing, subgroup 13-07F, except fuzzy kiwifruit Grape-0.037 Grape, juice = 0.2

Berry, low growing, subgroup 13-07G, except cranberry Strawberry-0.072

Grape, raisin 0.30 Grape, raisin = 1

The drinking water residue levels used in the dietary assessment were estimated using Tier I SCI-GROW (Screening Concentr ation in Groundwater) modeling. The Estimated Drinking Water Concentration (EDWC) fo r total toxic cyflufenamid residues are ground water residues based on applications to low gr owing berries (except cranberry), ornamentals, pome fruit, and small climbing fruit vines. The EDWC for tota l toxic residues is 4.68 for both the chronic and

cancer assessments. The water residues were dir ectly incorporated in the DEEM-FCID into the categories “water, direct, all sources” a nd “water, indirect, all sources.”

5.4.2 Percent Crop Treated Used in Dietary Assessment

The chronic dietary analysis assumed that 100% of the crop was tr eated (100% CT).

5.4.3 Acute Dietary Risk AssessmentPage 35 of 94 No toxic effects attributable to a single exposure to cyflufenamid have been identified; therefore, an acute reference dose (aRfD) has not been established and an acute dietary exposure

assessment was not conducted.

5.4.4 Chronic Dietary Risk Assessment

A partially refined chronic diet ary assessment was conducted. Th e assumptions of this dietary assessment included average field trial residues for all proposed crops. In addition, 100% crop treated (CT) was assumed. Empi rical processing factor s were used for apple juice and grape juice. A separate tolerance was set for grape, raisin; therefore, the processing factor for this commodity was set at 1. For all other pr ocessed commodities, DEEM version 7.81 default processing factors were assumed. A conser vative EDWC derived th rough EFED modeling which is unlikely to underestimate the concentr ation of cyflufenamid in drinking water was utilized. The ground water EDWC (4.68 u g/L) was incorporated directly into the dietary assessment (DP# 362954, R. Bohaty, 5/3/2010).

As shown in Table 5.4.6, the non-cancer chronic dietary (food and drin king water) exposure to cyflufenamid is below HED’s level of concern [i.e., <100% of the chr onic Population Adjusted Dose (cPAD)] for the general U.S. population an d all population subgroups. The chronic dietary exposure estimates are <1% of the cPAD for th e general U.S. population and 1% of the cPAD for all infants (<1 year old), the mo st highly exposed population subgroup.

5.4.5 Cancer Dietary Risk Assessment

Cyflufenamid is a likely human carcinogen and requi res a quantitative cancer risk assessment. The unit risk, Q 1 * = 6.61 X 10 -3 (mg/kg/day) -1 , of cyflufenamid is ba sed upon male mouse liver combined adenoma and carcinoma tumor rates. The ground water EDWC (4.68 u g/L) was incorporated directly into the dietary assessment (DP# 362954 , R. Bohaty, 5/3/2010). As shown in Table 5.4.6, the cancer dietary (food and dr inking water) exposure es timate for the general U.S. population is 9.5 x 10 -7 , which is below HED’s level of c oncern (generally in the range of

10 -6 ).

5.4.6 Summary Table

Table 5.4.6. Summary of Dietary (Food and Dri nking Water) Exposure a nd Risk for Cyflufenamid

.

Population Subgroup

cPAD (mg/kg/day) Chronic Dietary Cancer

Dietary Exposure (mg/kg/day) %

cPAD Dietary Exposure (mg/kg/day) Risk

Q* = 6.61 x 10 -3

General U.S. Population 0.2 0.000144 <1 0.000144 9.5 x 10 -7

All Infants (< 1 year old) 0.2 0.000423 1 1

NA 2 NA Children 1-2 years old 0.2 0.000330 <1

Children 3-5 years old 0.2 0.000271 <1Page 36 of 94 Table 5.4.6. Summary of Dietary (Food and Dri nking Water) Exposure a nd Risk for Cyflufenamid

.

Population Subgroup

cPAD (mg/kg/day) Chronic Dietary Cancer

Dietary Exposure (mg/kg/day) %

cPAD Dietary Exposure (mg/kg/day) Risk

Q* = 6.61 x 10 -3

Children 6-12 years old 0.2 0.000157 <1

Youth 13-19 years old 0.2 0.000097 <1

Adults 20-49 years old 0.2 0.000121 <1

Adults 50+ years old 0.2 0.000136 <1

Females 13-49 years old 0.2 0.000123 <1

1 The population subgroup with the highest estimated chronic dietary (food + drinking water) exposure and risk is indicated by bold text.

2 NA = not applicable

6.0 Residential (Non-Occupational) Exposure/Risk Characterization

6.1 Residential Handler Exposure

Although the proposed label for ornamental uses states “Not for homeowner use”, HED assessed residential handler exposure activities for the orna mental uses because there is a potential that homeowners could apply this product. HED reco mmends that the label language be revised to clarify the specific type of applicator and set ting (i.e., “applications ma de only by pest control operators in commercial and residential settings”).

Based on the use patterns, resident ial handlers could be exposed to cyflufenamid on a short-term

basis. The short-term dermal risk scenarios we re not necessary to assess because no hazard was identified for the dermal route of exposure. The short-term inhalation risk scenarios were assessed using the NOAEL of 5 mg/kg/day from the prenatal developmental study in rabbits. No inhalation absorption data are available; therefore, the toxicity by the inhalation route is considered to be equivalent to the estimated t oxicity by the oral route of exposure. For the cancer assessment, a dermal absorption factor of 12% was utilized in the dermal exposure component of the lifetime average da ily dose (LADD) while a Q1* of 6.61 x 10 -3 (mg/kg/day) -1
was used to estimate the risk. Also, a body wei ght of 70 kg was used in all exposure estimates because the endpoints were not ba sed on developmental effects.

HED assessed short-term non-cancer inhalation risk s and cancer risks for residential pesticide handlers in the following scenarios:

• mixing/loading/applying sprays with backpack sprayers,

• mixing/loading/applying sprays w ith low pressure handwand, and

• mixing/loading/applying sprays with hose-end sprayers.Page 37 of 94 Non-cancer risk estimates for re sidential handlers are included in Table 6.1.1. All inhalation

handler scenarios resulted in MO Es greater than the LOC (MOEs ≥ 100) and, therefore, are not

of concern.

Cancer risk estimates for residential handl ers are included in Ta ble 6.1.2. Assuming that residential applicators apply cyflufenamid twi ce a year per label inst ructions, all handler

scenarios resulted in cancer risks less than 1 x 10 -6 assuming typical clothing (e.g

. , short-sleeve shirt, short pants, and no gloves).

Table 6.1.1. Summary Of Short-Term Residential Handler Exposures And Risks

Exposure Scenario Crop Grouping Application

Rate a (lb ai/gal) Amount

Handled

Daily b
(gal/day) Baseline

Inhalation c

Unit Exposure (μg/lb ai) Baseline

Inhalation

Dose d
(mg/kg/day) Baseline

Inhalation

MOE e

(LOC ≥ 100)

Mixer/Loader/Applicator

Backpack (PHED) Nursery/Ornamentals 0.0011 5 30 0.0000024 2,100,000

Low Pressure

Handwand (ORETF) Nursery/Ornamentals 0.0011 5 3.8 0.00000031 16,000,000

Hose-End Sprayer (ORETF) Nursery/Ornamentals 0.0011 5 1.5 0.00000012 41,000,000

a Application rates are the maximum a pplication rates determined from proposed labels for cyflufenamid. Per label instructions and registrant correspondence: 0.0011 lb ai/gal = 3.4 oz conc/20 gal x 0.85 lb ai/gal x 1 gal/128 oz. b Amount handled per day values are HED estimates of amount handled per day based on ExpoSAC SOP No.12. c Inhalation: no respirator. d Inhalation dose = daily unit exposure ( μ g/lb ai) x application rate (lb a i/A) x area treated (A/day) x mg/1000 μ g x 100% inhalation absorption / body weight (70 kg adult). e Inhalation MOE = NOAEL (5.0 mg/kg/day) / inhalati on daily dose (mg/kg/day), where LOC = 100.Page 38 of 94Table 6.1.2. Summary Of Residential Handler Exposures and Cancer RisksExposure ScenarioCrop GroupingApplicationRatea(lb ai/gal)AmountHandledDailyb(gal/day)BaselineDermalcUnitExposure(mg/lb ai)BaselineInhalationdUnitExposure(μg/lb ai)BaselineDermalDosee(mg/kg/day)BaselineInhalationDosee(mg/kg/day)BaselineAverageDaily Dosef(mg/kg/day)ExposureDaysg(days/yr)BaselineLADDg(mg/kg/day)BaselineCancer RiskiMixer/Loader/ApplicatorBackpack(PHED)Nursery/Ornamentals 0.0011 5 5.1 30 0.000049 0.0000024 0.000052 2 2.0E-07 1.3E-09Low Pressure Handwand(ORETF)Nursery/Ornamentals 0.00115 56 3.8 0.00054 0.00000031 0.00054 2 2.1E-06 1.4E-08Hose-End Sprayer (ORETF)Nursery/Ornamentals 0.00115 39 1.5 0.00038 0.00000012 0.00038 2 1.5E-06 9.8E-09a Application rates are the maximum application rates determined from proposed labelsfor cyflufenamid. Per label instructionsand registrant correspondence: 0.0011 lbai/gal = 3.4 oz conc/20 gal x 0.85 lb ai/gal x 1 gal/128 oz.b Amount handled per day values areHED estimates of amount handled perday based on ExpoSAC SOP No.12.c Dermal: short pants, short sleeves, and no gloves.d Inhalation: no respirator.e Dose = daily unit exposure (μg/lb ai) x application rate (lb ai/A) x area treated (A/day) x mg/1000μg (as needed) x100% inhalation absorption or 12% dermalabsorption / body weight (70 kg adult).f Average daily dose (ADD) (mg/kg/day) = dermal dose (mg/kg/day)+ inhalation dose (mg/kg/day).g Exposure days: Based on label instructions: maximum of 2 applications per year.h Lifetime average daily dose (LADD) (mg/kg/day) =ADD x (2 days/yr x 50 yr) / (365 days/yr x 70 yr).i Cancer risk = LADD (mg/kg/day) x Q* of 6.61 x 10-3(mg/kg/day)-1.Page 39 of 94 6.2 Post-Application Exposure

A suspension concentrate liquid is proposed for use at a maximum of two applications per year to ornamental plantings and trees in residential areas and recreationa l sites. When determining the potential for residential exposure, the Agency cons iders foliar residues, leaf to skin/hand residue transfer, children’s hand-to-mout h residue transfer, and exposure time. In the case of cyflufenamid, potential exposure to adults and chil dren would be negligible for the fo llowing reasons: 1) Activities such as pruning/thinning orname ntals or playing in and around or namentals when residues may be present on the day of the application ar e unlikely to co-occur; 2) if presen t, leaf to skin residue transfer would be negligible because of the minimal fr equency and duration of contact; 3) children young enough to exhibit hand-to-mouth behavior would not typi cally play in ornamental beds or trees. Based on the frequency of application and unlikely poten tial for postapplication exposure, residential postapplication risks were not quantitatively assessed; thus, there are no postappl ication residential risk concerns for this use pattern.

Based on the Agency's current practices, a qua ntitative postapplication inhalation exposure assessment was not performed for cyflufenamid at this time. However, volatilization of pesticides may be a potential source of postappli cation inhalation exposure to individuals nearby to pesticide applications. The Agency sought expert advice and input on issues related to volatilization of pesticid es from its Federal Insecticide, Fungicide, and Rodenticide Act Scientific Advisory Panel (SAP) in December 2009. The Agency received the SAP’s final report on March 2, 2010 (http://www.epa.gov/scipol y/SAP/meetings/2009/120109meeting.html ). The Agency is in the process of ev aluating the SAP report and may, as appropriate, develop policies and procedures to identify the need for and, s ubsequently, the way to in corporate postapplication inhalation exposure into the Agency 's risk assessments. If new policies or procedures are put into place, the Agency may revisit the need fo r a quantitative postapplic ation inhalation exposure assessment for cyflufenamid.

6.3 Combined Exposure

Residential postapplication pestic ide exposure can occur within a scenario from multiple routes of exposure (e.g., dermal, inhalation, and oral). These exposures to pesticides do not occur as single, isolated events, but rather as a series of sequential or c oncurrent events that may overlap or be linked in time and space. Based on this, ri sk estimates resulting from different exposure routes may be combined when it is likely that they can occur simultaneously based on the use pattern and when the toxicological effects across different routes of exposure are the same. If appropriate for an exposure scenario, the re commended routes of exposure for combining include dermal, inhalation, and hand-to-mouth.

For the non-cancer assessment, combined exposures were not necessary to assess for cyflufenamid since inhalation e xposures for the handlers were the only scenario quantitatively assessed. (As discussed in Sect ion 6.1, no dermal hazard was iden tified.) However, for thePage 40 of 94 cancer assessment, dermal and in halation exposure were combined in order to estimate the

LADD and associated risk.

6.4 Spray Drift

Spray drift is always a potential source of expos ure to residents nearby to spraying operations. This is particularly the case with aerial appl ication, but, to a lesser ex tent, could also be a potential source of exposure from the ground ap plication method employed for cyflufenamid. The Agency has been working with the Spray Dr ift Task Force, EPA Regional Offices and State Lead Agencies for pesticide regulation and ot her parties to develop the best spray drift management practices (see the Agency’s Spra y Drift website for more information at http://www.epa.gov/opp00001/factsheets/ spraydrift.htm). On a chemical by chemical basis, the Agency is now requiring interim mitigation measures for aerial applications that must be placed on product labels/labeling. The Agency has comp leted its evaluation of the new database submitted by the Spray Drift Task Force, a membership of U.S. pesticide registrants, and is developing a policy on how to appr opriately apply the data and th e AgDRIFT computer model to its risk assessments for pesticides applied by ai r, orchard airblast and ground hydraulic methods. After the policy is in place, the Agency may impose further refinements in spray drift management practices to reduce off-target drif t with specific products with significant risks

associated with drift.

7.0 Aggregate Exposure/Risk Characterization

Under FQPA, HED must consider and aggregate pesticide exposures and risk from three major sources: food, drinking water, and residential exposures.

7.1 Acute Aggregate Risk

Acute exposures were not considered because an a dverse effect attributable to a single dose was

not identified.

7.2 Short- and Intermediate-Term Aggregate Risk

There is potential short- and in termediate-term exposure to cyflufenamid via the dietary pathway (which is considered background exposure) and the residential pathway (which is considered the primary pathway). Since no hazard has been id entified for the dermal route of residential exposure, the aggregate exposure assessment fo r adults includes di etary (food and drinking water) and inhalation exposures. For childre n, exposure includes only food and drinking water since 1) no dermal hazard was identified; 2) or namentals are the only residential use; and 3) dermal, inhalation, and incidental oral expos ure to children from treated ornamentals is considered to be negligible. As shown in Ta ble 7.2, all aggregate MOEs are greater than 100; therefore, aggregate exposu re is not of concern.Page 41 of 94 Table 7.2. Short-Term and Intermediate-T erm Aggregate Risk Calculations

Population Short- and Intermediate-Term Scenario

NOAEL mg/kg/day LOC 1 Max
Allowable Exposure 2
(mg/kg/day) Average

Food &

Water Exposure 3
(mg/kg/day) Residential Exposure 4
(mg/kg/day) Aggregate MOE (food and residential) 5

Adult Male 5 100 0.05 0.000144 0.0000024 34,000

Adult Female 5 100 0.05 0.000123 0.0000024 40,000

Child 5 100 0.05 0.000330 NA 15,000

1 This is based on the conventional uncertainty factor of 100x (10x for interspecies extrapolation and 10x for intraspecies variation).

2 Maximum Allowable Exposure (mg/kg/day) = NOAEL/LOC.

3 Dietary exposures for these populations are based on the following population subgroups: Adult Male: U.S. population; Adult Female: Females 13-49 years old; and Child: Children 1-2 years old.

4 Since no dermal hazard was identified, residential exposure (RE) = oral exposure + inhalation exposure (Table 6.1.1). RE adult = inhalation exposure since no dermal hazard was identified. RE child is not applicable since no dermal hazard was identified, and inhala tion and incidental oral exposures are negligible from use on ornamentals (the only residential use)

. 5 Aggregate MOE = [NOAEL ÷ (avg food & water exposure + residential exposure)].

7.3 Chronic Aggregate Risk

There is potential chronic exposure via the diet (food and drinking water). Refer to Section 5.4.

7.4 Cancer Aggregate Risk

As shown in Table 7.4 below, the aggregate cance r risk (food, water, and residential) is 9.7 x

10 -7 . The unit risk for cyflufenamid, Q 1 * , is 6.61 x 10 -3 (mg/kg/day) -1 . The Q 1 * is based upon

male mouse liver combined adenoma and carcino ma tumor rates and is expressed in human equivalents.

Table 7.4 Aggregate Cancer Calculations

Population Q 1 * (mg/kg/day )-1 Negligible Risk

Level Chronic Food & Water Exposure (mg/kg/day) Residential Exposure (LADD) 1 Aggregate

Cancer Risk (food, water and residential) 2
U.S. Pop 6.61 x 10 -3 generally in the range of 10 -6 0.000144 2.1 x 10 -6 9.7 x 10 -7

1 The residential exposure value used in the ag gregate assessment is taken from Table 6.1.2.

2 Aggregate Risk Cancer = Q 1 * x [Chronic Food & Water Exposure + Re sidential Exposure (Lifetime Average Daily Dose)]Page 42 of 94 8.0 Cumulative Exposure/Risk Characterization

Unlike other pesticides for which EPA has follo wed a cumulative risk approach based on a common mechanism of toxicity, EPA has not made a common mechanism of toxicity finding as to cyflufenamid and any other substances and cy flufenamid does not appear to produce a toxic metabolite produced by other substances. For the purposes of this tolerance action, therefore, EPA has not assumed that cyflufenamid has a common mechanism of toxicity with other substances. For information regarding EPA’s efforts to determine which chemicals have a common mechanism of toxicity and to evaluate th e cumulative effects of such chemicals, see the policy statements released by EPA’s Office of Pesticide Programs concerning common mechanism determinations and proc edures for cumulating effects from substances found to have

a common mechanism on EPA’s website at http://www.epa.gov/pestic ides/cumulative/.

9.0 Occupational Exposure/Risk Characterization

Occupational exposures are expected to occur from the proposed cyflufenamid uses on cucurbits, grapes, pome fruit, strawberries, and ornament als. Short- (1-30 consecutive days) and intermediate-term (1-6 consecuti ve months) dermal and inhalati on exposures are expected from handler activities while only short- and intermed iate-term dermal exposures are expected from postapplication activities.

The short- and intermediate-term dermal risk sc enarios were not necessary to assess because no

hazard was identified for the dermal route of exposure. The short- and intermediate-term

inhalation risk scenarios were assessed usi ng the NOAEL of 5 mg/kg/da y from the prenatal developmental study in rabbits. No inhalation ab sorption data are availa ble; therefore, the toxicity by the inhalation route is considered to be equivalent to the estimated toxicity by the oral route of exposure. For the cancer assessment, a de rmal absorption factor of 12% was utilized in the dermal exposure component of the lif etime average daily dose (LADD) while a Q 1 * of 6.61 x

10 -3 (mg/kg/day) -1 used was to estimate the risk. Als o, a body weight of 70 kg was used in all exposure estimates because the endpoints we re not based on developmental effects.

9.1 Short-/Intermediate-/Long-Term/Cancer Handler Risk

Chemical-specific exposure data were not submitted to support the proposed cyflufenamid uses. When chemical-specific monitoring data are not av ailable, it is the policy of HED to use data from PHED Version 1.1 as presented in the PHED Surrogate Exposure Guide (8/98) 1 or ORETF to assess handler exposures for regulatory actio ns. Additionally, typical HED standard values were used for the amount treated per day (ExpoSAC Policy No. 9, 07/05/2000).

1 HED Science Advisory Council for Exposure (ExpoSAC) Policy .007, “Use of Values from PHED Surrogate Table and Chemical-Specific Data” HED, OPP, 1/28/99Page 43 of 94 Based upon the proposed cyflufenamid use patterns, HED assessed risks to occupational pesticide handlers in the following scenarios:

• mixing/loading liquid concentrates to support aerial applications,

• mixing/loading liquid concentrates to support airblast applications,

• mixing/loading liquid concentrates to support groundboom applications,

• applying sprays with aircraft,

• applying sprays with airblast equipment,

• applying sprays with groundboom equipment,

• flagging to support aerial spray applications,

• mixing/loading/applying sprays with backpack sprayers,

• mixing/loading/applying sprays with low pressure handwand.

Non-cancer risk estimates for o ccupational handlers are included in Table 9.1.1. All inhalation

handler scenarios resulted in MO Es greater than the LOC (MOEs ≥ 100) and, therefore, are not

of concern.

Cancer risk estimates for occupational handlers are included in Table 9.1.2. The cancer risks for

some of the handler scenarios were in the 10 -6 range at the baseline leve l of clothing. However,

all handler scenarios resulted in cancer risks less than 1 x 10 -6 when wearing additional PPE (e.g

. , long-sleeve shirt, long pants, shoes, socks, a nd chemical resistant gloves) as required by the

label.

Table 9.1.1. Summary Of Short-and Intermediate -Term Occupational Handler Exposures And Risks

Exposure Scenario Crop Grouping Application

Rate a (lb ai/A) Area

Treated Daily b
(acres/day) Baseline

Inhalation c

Unit Exposure (μg/lb ai) Baseline

Inhalation

Dose d
(mg/kg/day) Baseline

Inhalation

MOE e

(LOC ≥ 100)

Mixer/Loader Mixing/Loading Liquid

Concentrates for

Aerial Applications Cucurbits 0.011 350 1.2 0.000066 76,000

Pome fruit

Grapes

Strawberries 0.022 350 1.2 0.00013 38,000

Mixing/Loading Liquids

Concentrates for

Groundboom Applications Cucurbits 0.011 80 1.2 0.000015 330,000

Strawberries Nursery/ornamentals 0.022 80 1.2 0.000030 170,000

Mixing/Loading Liquids

Concentrates for

Airblast Applications Pome fruit Grape Nursery/ornamentals 0.022 40 1.2 0.000015 330,000

Applicator

Applying Sprays Cucurbits 0.011 350 0.068 0.0000037 1,300,000Page 44 of 94 Table 9.1.1. Summary Of Short-and Intermediate -Term Occupational Handler Exposures And Risks

Exposure Scenario Crop Grouping Application

Rate a (lb ai/A) Area

Treated Daily b
(acres/day) Baseline

Inhalation c

Unit Exposure (μg/lb ai) Baseline

Inhalation

Dose d
(mg/kg/day) Baseline

Inhalation

MOE e

(LOC ≥ 100)

via Aerial Equipment (enclosed cockpit) Pome fruit Grapes

Strawberries 0.022 350 0.068 0.0000075 670,000

Applying Sprays

via Groundboom Equipment (open cab) Cucurbits 0.011 80 0.74 0.0000093 540,000

Strawberries Nursery/ornamentals 0.022 80 0.74 0.000019 270,000

Applying Sprays

via Airblast Equipment (open cab) Pome fruit Grapes Nursery/ornamentals 0.022 40 4.5 0.000057 88,000

Flagger

Flagging for Aerial Sprays Applications Cucurbits 0.011 350 0.35 0.000019 260,000

Pome fruit

Grapes,

Strawberries 0.022 350 0.35 0.000039 130,000

Mixer/Loader/Applicator

Backpack Nursery/Ornamentals 0.0011 f 40 30 0.000019 260,000

(lb ai/gal) (gal/day)

Low Pressure

Handwand Nursery/Ornamentals 0.0011 f 40

30 0.000019 260,000 (lb ai/gal) (gal/day)

a Application rates are the maximum application rate s determined from proposed labels for cyflufenamid. b Amount handled per day values are HED estimates of acre s treated per day based on Exposure SAC SOP #9 “Standard Values for Daily Acres Trea ted in Agriculture,” c Baseline Inhalation: no respirator. d Inhalation dose = daily unit exposure ( μ g/lb ai) x application rate (lb a i/A) x area treated (A/day) x mg/1000 μ g x 100% inhalation absorption / body weight (70 kg adult). e Inhalation MOE = NOAEL (5.0 mg/kg/day) / inhala tion daily dose (mg/kg/day) , where LOC = 100 f Per label instructions and registrant correspondence: 0.0011 lb ai/gal = 3.4 oz conc/20 gal x 0.85 lb ai/gal x 1 gal/128

ozPage 45 of 94Table 9.1.2. Summary Of PPE Occupational Handler Exposures and Cancer RisksExposure ScenarioCrop GroupingApplicationRatea(lb ai/A)AreaTreatedDailyb(acres/day)PPEDermalcUnitExposure(mg/lb ai)BaselineInhalationdUnitExposure(μg/lb ai)PPEDermalDosee(mg/kg/day)BaselineInhalationDosee(mg/kg/day)AverageDaily Dosef(mg/kg/day)ExposureDaysg(days/yr)LADDg(mg/kg/day)Cancer RiskiMixer/LoaderMixing/Loading LiquidConcentrates for AerialApplicationsCucurbits 0.011 350 0.023 1.2 0.00015 0.000066 0.00022 30 0.0000090 5.9E-08Pome fruitGrapesStrawberries0.022 350 0.023 1.2 0.00030 0.00013 0.00044 30 0.000018 1.2E-07Mixing/Loading LiquidsConcentrates forGroundboom ApplicationsCucurbits 0.011 80 0.023 1.2 0.000035 0.000015 0.000050 30 0.0000020 1.4E-08StrawberriesNursery/ornamentals0.022 80 0.023 1.2 0.000069 0.000030 0.00010 30 0.0000041 2.7E-08Mixing/Loading LiquidsConcentrates for AirblastApplicationsPome fruitGrapesNursery/ornamentals0.022 40 0.023 1.2 0.000035 0.000015 0.000050 30 0.0000020 1.4E-08ApplicatorApplying Sprays via AerialEquipment (enclosedcockpit)Cucurbits 0.011 350 0.0022 0.068 0.000015 0.0000037 0.000018 30 0.00000075 5.0E-09Pome fruitGrapesStrawberries0.022 350 0.0022 0.068 0.000029 0.0000075 0.000037 30 0.0000015 9.9E-09Applying Sprays viaGroundboom Equipment(open cab)Cucurbits 0.011 80 0.014 0.74 0.000021 0.0000093 0.000030 30 0.0000013 8.3E-09StrawberriesNursery/ornamentals0.022 80 0.014 0.74 0.000042 0.000019 0.000061 30 0.0000025 1.7E-08Applying Sprays via AirblastEquipment (open cab)Pome fruitGrapesNursery/ornamentals0.022 40 0.24 4.5 0.00036 0.000057 0.00042 30 0.000017 1.1E-07FlaggerFlagging for Aerial SpraysApplicationsCucurbits 0.011 350 NA 0.35 NA 0.000019 NA 30 NA NAPome fruitGrapesNursery/ornamentals0.022 350 NA 0.35 NA 0.000039 NA 30 NA NAMixer/Loader/ApplicatorPage 46 of 94Table 9.1.2. Summary Of PPE Occupational Handler Exposures and Cancer RisksExposure ScenarioCrop GroupingApplicationRatea(lb ai/A)AreaTreatedDailyb(acres/day)PPEDermalcUnitExposure(mg/lb ai)BaselineInhalationdUnitExposure(μg/lb ai)PPEDermalDosee(mg/kg/day)BaselineInhalationDosee(mg/kg/day)AverageDaily Dosef(mg/kg/day)ExposureDaysg(days/yr)LADDg(mg/kg/day)Cancer RiskiBackpackNursery/Ornamentals0.0011j402.5 30 0.00019 0.000019 0.00021 30 0.0000087 5.8E-08(lb ai/gal) (gal/day)Low Pressure HandwandNursery/Ornamentals0.0011j400.43 30 0.000033 0.000019 0.000053 30 0.0000022 1.4E-08(lb ai/gal) (gal/day)(lb ai/gal) (gal/day)a Application rates are the maximum application rates determined from proposed labels for cyflufenamid.b Amount handled per day values are HED estimates of acres treated per day based on ExpoSAC SOP No.9 “Standard Values for DailyAcres Treated in Agriculture,”c PPE Dermal: long pants, long sleeves, and chemical resistant glovesd Baseline Inhalation: no respirator.e Dose = daily unit exposure (μg/lb ai) x application rate (lb ai/A) x area treated (A/day) x mg/1000μg (as needed) x 100% inhalation absorption or 12% dermal absorption / bodyweight (70 kg adult).f Average daily dose (ADD) (mg/kg/day) = dermal dose (mg/kg/day)+ inhalation dose (mg/kg/day)g Exposure days: HED default for number of worker daysh LADD (Lifetime average daily dose) (mg/kg/day) = ADD x (30 days/yr x 35 yr) / (365 days/yr x 70 yr)i Cancer risk = LADD (mg/kg/day) x Q* of 6.6 x 10-3(mg/kg/day)-1j Per label instructions and registrant correspondence: 0.0011 lbai/gal = 3.4 oz conc/20 gal x 0.85 lb ai/gal x 1 gal/128 ozPage 47 of 94 9.2 Short-/Intermediate-/Long-Term /Cancer Post-Application Risk

There is a possibility for agri cultural workers to have posta pplication dermal exposure to cyflufenamid following its proposed new uses. However, non-cancer occupational postapplication dermal exposure and risks were not assessed because, as previously discussed in Section 2.0, no hazard was identified for the dermal route of exposure.

Because cyflufenamid is “likely to be carcinogenic to humans”, cancer risks from postapplication activities were a ssessed. Chemical-specific data relevant to postapplication exposure (i.e., dislodgeable foliar residue (DFR) data) for the new cyflufenamid proposed uses were not submitted. Therefore, postapplication exposure estimates were calculated using the standard HED ExpoSAC assumptions (i.e., defau lt DFR input parameters and exposure duration) and HED transfer coefficients (SOP No. 3.1).

HED uses a concept known as the transfer coefficient (TC) to numerically represent the postapplication exposures one would r eceive (generally presented as cm 2 /hour). A TC is a

measure of the residue transferred from a treat ed surface to a person who is doing a task or activity in a treated area. These values are the ratio of an e xposure for a given task or activity to the amount of pesticide residue on treated surfaces av ailable for transfer. HED has developed a series of standard TCs that are unique for a variety of job tasks or activities that are used in lieu of chemical- and scenario-specific data. HED used Policy 3.1 Science Advisory Council for Exposure Policy Regarding Agricu ltural Transfer Coefficients to estimate TCs for the various tasks that postapplication workers might perform on each crop grouping.

In the absence of chemical-specific DFR data, HED assumes that 20% of the initial application is available for transfer on day 0 (i.e ., 12 hours after application) and th at the residue dissipates at a rate of 10% per day. Furthermore, HED also assumes an 8 hour workday for postapplication workers. Average daily doses (ADD) were cal culated on each day afte r application. LADDs and cancer risks were calculated for each postapplication activity.

Table 9.2 presents the results of the occupati onal postapplication cancer risk assessment when HED default assumptions are used. Using surro gate data, the cancer risks from agricultural postapplication activities range from 7.4 x 10 -8 to 4.1 x 10 -6 on day 0 (12 hours following application). Although cyflufenamid specific DFR studies on grapes and ornamentals could be used to refine and mitigate the cancer risks for these scenarios, this assessment incorporates conservative parameters such as maximum applic ation rates and assuming workers are exposed to “Day 0” cyflufenamid residue levels for 250 days/year even though onl y two applications per ornamental type per year can be made with a retreatment interval of 21 days.Page 48 of 94Table 9.2. Summary of Occupational Postapplication Cancer Risks for Proposed Cyflufenamid Uses*CropAppl.Rate(lb ai/A)Fraction ofai Retainedon Foliage1TransferCoefficient(cm2/hr)2DislodgeableFoliarResidue3(μg/cm2)AverageDaily Dose4(mg/kg/day)LADD5(mg/kg/day)CancerRisk6Pome fruit (fruittrees)0.022 20%High/full: harvesting (1,500)0.049 0.0010 0.000042 2.8E-07High/full: thinning (3,000)0.049 0.0020 0.000083 5.5E-07Strawberries 0.022 20%Low/full: irrigation, mulching, scouting, hand weeding(400)0.049 0.00027 0.000011 7.4E-08Low/full: hand harvest, hand pruning, pinching, training(1,500)0.049 0.0010 0.000042 2.8E-07Grapes 0.022 20%High/full: hand weeding (500)0.049 0.00034 0.000014 9.2E-08High/full: scouting (1,000)0.049 0.00068 0.000028 1.8E-07High/full: hand harvest, hand pruning, thinning (5,000) 0.049 0.0034 0.00014 9.2E-07High/full: girdling, cane turning, cane tying (10,000) 0.026 0.0068 0.00028 1.8E-06Cucurbits 0.011 20%Low/full: irrigation, scouting, hand weeding (1,500) 0.025 0.00051 0.000021 1.4E-07Low/full: hand harvest, hand pruning, thinning (2,500) 0.025 0.00085 0.000035 2.3E-07Ornamentals0.022 20%High/full: pruning, tying (110) 0.049 0.00062 0.000025 1.7E-07High/full: moving potted plants(400) 0.049 0.0023 0.000093 6.1E-07High/full: cut flowers (2,700) 0.049 0.0018 0.00063 4.1E-06* Unless otherwise noted, all exposures and risks are estimated on day 0 after application1HED default2Transfer coefficients are taken from the HED ExpoSAC SOP 3.1 (August 2000)3Dislodgeable Foliar ResiduePostapplication day(μg/cm2) = Application rate (lb ai/A) x Fractionof ai Retained on the Foliage x (1- Fraction of Residue that Dissipates Daily)postapplication day zerox 4.54E+8 μg/lbx 2.47E-8 A/cm24Average Daily Dose = [Dislodgeable Foliar Residuex 0.001 mg/μg x Dermal Transfer Coefficient (cm2/hr) x Exposure Time (8 hr/day) x Dermal absorption (12%)]/Body weight (70 kg)5LADD = ADDx (30 or 250 days/yr x 35 yr) / (365 days/yr x 70 yr); note: itwas assumed that exposure frequency is 30 days/yr for all scenarios except “cut flowers” where 250days/yr was used6Cancer risk = LADD (mg/kg/day) x Q* (mg/kg/day)-1Page 49 of 94 Based on the Agency's current practices, a qua ntitative postapplication inhalation exposure assessment was not performed for cyflufenamid at this time. However, volatilization of pesticides may be a potential source of postappli cation inhalation exposure to individuals nearby to pesticide applications. The Agency sought expert advice and input on issues related to volatilization of pesticid es from its Federal Insecticide, Fungicide, and Rodenticide Act Scientific Advisory Panel (SAP) in December 2009. The Agency received the SAP’s final report on March 2, 2010 (http://www.epa.gov/scipol y/SAP/meetings/2009/120109meeting.html ) and is in the process of evaluating the SAP report. The Agency may, as appr opriate, develop policies and procedures to identify the need for and, s ubsequently, the way to in corporate postapplication inhalation exposure into the Agency 's risk assessments. If new policies or procedures are put into place, the Agency may revisit the need fo r a quantitative postapplic ation inhalation exposure assessment for cyflufenamid.

Restricted Entry Interval: For all activitie s and crops, the REI of 12 hours on the proposed label is adequate to protect workers. The REI is based on the acute toxicity of cyflufenamid technical material. Cyflufenamid is classified in acute toxicity category III/IV for acute dermal toxicity and category IV/III for pr imary eye irritation and primary skin irritation. It is not a dermal sensitizer. Under the WPS for Agricultur al Pesticides, active ingredients classified as acute toxicity categories III or IV for these rout es are assigned a 12-hour REI. Therefore, the

REI of 12 hours for all activities and crops is appropriate and meets the requirements of the

WPS.

10.0 Incidents

No information is available from use in the U.S. since this is the first registration in the U.S.

11.0 Environmental Justice Considerations

Potential areas of environmental ju stice concerns, to the extent possi ble, were considered in this human health risk assessment, in accordance w ith U.S. Executive Order 12898, "Federal Actions

to Address Environmental Just ice in Minority Populations an d Low-Income Populations," http://www.eh.doe.gov/oepa/ guidance/justice/eo12898.pdf ).

As a part of every pesticide risk assessment , OPP considers a large variety of consumer subgroups according to well-esta blished procedures. In line with OPP policy, HED estimates risks to population subgroups fr om pesticide exposures that ar e based on patterns of that subgroup’s food and water consumption, and activ ities in and around the home that involve pesticide use in a residential se tting. Extensive data on food consumption patterns are compiled by the USDA under the Continuing Survey of Food Intake by Individuals (CSFII) and are used in pesticide risk assessments for all registered fo od uses of a pesticide. These data are analyzed and categorized by subgroups based on age, season of the year, ethnic group, and region of the country. Additionally, OPP is able to assess di etary exposure to smaller, specialized subgroups and exposure assessments are performed when c onditions or circumstances warrant. Whenever appropriate, non-dietary exposures based on home use of pesticide products and associated risks for adult applicators and for t oddlers, youths, and adults enteri ng or playing on treated areasPage 50 of 94 postapplication are evaluated. Further considerat ions are currently in development as OPP has committed resources and expertise to the developm ent of specialized software and models that consider exposure to bystanders and farm worker s as well as lifestyle and traditional dietary patterns among specific subgroups.

12.0 References

Kramer, G.F., Ph.D., 5/18/2010, DP#377139, Cyflufenamid. Report of the Residues of Concern Knowledge-based Subcommittee (ROCKS).

Brunsman, L.L., 4/8/ 2010, DP#363403, TXR#0055318, Cyflufenamid Qualitative Risk

Assessment Based on CD Rat and CD -1 Mouse Dietary Studies.

Negussie, M., 7/23/2010, DP#364556, Cyflufenamid. Section 3 Registration Request for Uses on Cucurbit Vegetables (Crop Group 9), Pome Fruit (Crop Group 11), Small Fruit Vine Climbing Except Fuzzy Kiwifruit (Subgroup 13-07F), and Low Growing Berry (Subgroup 13-07G), except Cranberry. Summary of Analytical Chemistry and Residue Data.

Negussie, M., 7/20/2010, DP#364559, Cyflufenamid: Chronic Aggregate Dietary (Food and Drinking Water) Exposure and Risk Assessments for the First Food Uses in Support of a Section 3 Registration Action on Cucurbit Vegetables (Crop Group 9), Pome Fruit (Crop Group 11), Small Fruit Vine Climbing Except Fuzzy Kiw ifruit (Subgroup 13-07F), and Low Growing Berry (Subgroup 13-07G), except Cranberry

.

Walls, C., Ph.D., 5/24/10, DP#364557, C YFLUFENAMID: Occupational and Residential Exposure/Risk Assessment for Use on Cucurbit Ve getables, Pome Fruit, Small Fruit Vine, Low Growing Berry, and Ornamentals

.

Bohaty, R.F.H., Ph.D., 4/15/2010, DP#360795, The Drinking Water Degradation Products Identification Memorandum for Cyflufenamid.

Bohaty, R.F.H., Ph.D., 5/3/2010, DP#362954, Drinking Water Assessment for the Registration of Cyflufenamid (MILTREX ®

10 SC FUNGICIDE) Uses on: Cucurbit Vegetables (Crop Subgroup 9), Low Growing Berries (Crop Subgroup 13-07G), Outdoor Landscape and Nursery Ornamentals, Pome Fruit (Crop Group 11), and Small Fruit Vines Climbing except Fuzzy Kiwifruit (Crop Subgroup 13-07F).Page 51 of 94 Appendix A. Toxicology Prof ile and Executive Summaries

A.1 Toxicology Data Requirements The requirements (40 CFR 158.340) for food uses for cy flufenamid are in Table 1. Use of the new guideline numbers does not imply that the new (1998) guideline protocols were used. A description of the outstanding study is in Appendix E.

Table 1.: Toxicolo gy Data Requirements

Test Technical

Required Satisfied

870.1100 Acute Oral Toxicity ....................................................... 870.1200 Acute Dermal Toxicity ..................................................

870.1300 Acute In halation Toxicity ..............................................

870.2400 Primar y Eye Irr itation

.................................................... 870.2500 Primar y Dermal Irr itation

.............................................. 870.2600 Derm al Sensiti zation...................................................... yes

yes

yes

yes

yes

yes yes

yes

yes

yes

yes

yes

870.3100 Oral Su bchronic (rodent) ...............................................

870.3150 Oral Su bchronic (nonr odent) ......................................... 870.3200 21-Day Dermal .............................................................. 870.3250 90-Day Dermal ..............................................................

870.3465 90-D ay Inhala tion .......................................................... yes

yes

yes

cr

cr yes

yes

yes

no

no

870.3700a Developmental Toxicity (rodent) ................................... 870.3700b Developmental Toxicity (nonrodent).............................

870.3800 Reproduc tion ................................................................. yes

yes

yes yes

yes

yes

870.4100a Chronic T oxicity (rodent) ..............................................

870.4100b Chronic Toxi city (nonrodent) ........................................ 870.4200a Oncoge nicity (ra t) .......................................................... 870.4200b Oncogenicity (mouse) ....................................................

870.4300 Chroni c/Oncogeni city .................................................... na

yes

na

yes

yes -
yes

-
yes

yes

870.5100 Mutagenicity—G ene Mutation - bacterial ..................... 870.5300 Mutagenicity—Gene Mutation - mammalian ................ 870.5375 Mutagenicity—Struc tural Chromosomal Aberrations ... 870.5395 Mutagenicity—O ther Genotoxic Effects ....................... yes

yes

yes

yes yes

yes

yes

yes

870.6100a Acute Delaye d Neurotox. (hen) ..................................... 870.6100b 90-Day Neurotoxicity (hen) ...........................................

870.6200a Acute Neurotox. Screening Battery (rat) ....................... 870.6200b 90-Day Neuro. Screening Batte ry (rat) .......................... 870.6300 Develop. Neuro .............................................................. no

no

yes

yes

cr -

-

no +

yes

no

870.7485 Gene ral Metabo lism ......................................................

870.7600 Derm al Penetr ation ........................................................ 870.7800 Immunotoxicity ............................................................. yes

cr

yes yes

no

yes

+Waiver was deniedPage 52 of 94 A.2 Toxicity Profiles

Table A.2.1 Acute Toxicity Pr ofile – Cyflufenamid, TGAI

Guideline No. Study Type MRID(s) Results Toxicity Category

870.1100 Acute oral [rat] 47620418 LD 50 > 5000 mg/kg IV

870.1200 Acute dermal [rat] 47620419 LD 50 > 2000 mg/kg III

870.1300 Acute inhalation [rat] 47620420 LC 50 > 4.76 mg/L IV

870.2400 Acute eye irritation [rabbit] 47620421 minimally irritating III

870.2500 Acute dermal irritation [rabbit] 47620422 Non-irritating IV

870.2600 Skin sensitization [guinea pig] 47620423 Negative NA

Table A.2.2 Acute Toxicity Profil e – Cyflufenamid Metabolites

Guideline No. Acute Oral Toxicity in Rats,

Metabolite: MRID(s) Results Toxicity Category

870.1100 149-F 47620609 LD 50 = 2828 mg/kg III

870.1100 149-F1 47620610 LD 50 = 394 mg/kg II

870.1100 149-F6 47620611 LD 50 = 686 mg/kg III

870.1100 149-F11 47620612 LD 50 = 1162 mg/kg III

870.1100 149-( E )-FB 47620613 LD 50 > 5000 mg/kg IVPage 53 of 94 Table A.2.2 Subchronic, Chronic and Other Toxicity Profile

Guideline

No. Study Type MRID No. (year)/

Classification /Doses Results

870.3100 90-Day oral toxicity (rat) 47638204 (1997) Acceptable/guideline 0, 50, 300, 1800 or 10800

ppm M: 0, 3.3, 20.1, 117.4, 673.3 mg/kg/day F: 0, 4.1, 24.7, 143.9, 782.6 mg/kg/day NOAEL = 20.1 mg/kg/day ♂ , 24.7 mg/kg/day ♀ LOAEL = 117.4 mg/kg/day ♂ , 143.9 mg/kg/day ♀ , based on decreased body weight gain, centrilobular hepatocellular hypertrophy and increased relative organ weights (liver, kidney, thyroid)

870.3100 90-Day oral toxicity (mouse) 47638202 (1998) Acceptable/guideline 0, 100, 400, 1600 or 7000

ppm M: 0, 14.0, 50.7, 218.1 or 808.0 mg/kg/day

F: 0, 17.6, 70.8, 295.1 or 940.2 mg/kg/day NOAEL = 218.1 mg/kg/day ♂ , 295.1 mg/kg/day ♀

LOAEL = 808.0 mg/kg/day ♂ , 940.2 mg/kg/day ♀ based on hepatocellular hypertrophy, increased liver weight.

870.3150 90-Day oral toxicity (dog) 47620504 (1999) Acceptable/guideline 0, 150, 500, 1500 ppm M: 0, 6.5, 23.2, 76.2 mg/kg/day

F: 0, 7.5, 24.4, 70.5 mg/kg/day NOAEL = 23.2 mg/kg/day ♂ , 24.4 mg/kg/day ♀
LOAEL = 76.2 mg/kg/day ♂ , 70.5 mg/kg/day ♀ based on decreased body weight gain and hepatomegaly accompanied by vacuolated hepatocytes and fat deposition

870.3200 21/28-Day dermal toxicity (rat) 47620428 (2000) Acceptable/guideline M/F: 0, 100, 300, 1000 mg/kg/day NOAEL = 1000 mg/kg/day. LOAEL > 1000 mg/kg/day.

870.3700a Prenatal developmental in (rat) 47620506 (2000) Acceptable/guideline F: 0, 100, 300, 1000 mg/kg/day Maternal NOAEL = 300 mg/kg/day LOAEL = 1000 mg/kg/day based on increased liver weight and brown staining of coat in anterior region, piloerection and ungroomed appearance, indicators of deteriorating health Developmental NOAEL = 1000 mg/kg/day. LOAEL > 1000 mg/kg/day.

870.3700b Prenatal developmental in (rabbit) 47620508 (2000) Acceptable/guideline F: 0, 10, 60, 300 mg/kg/day

47620509 (2001) Acceptable/non-guideline F: 0, 5, 10 mg/kg/day Maternal NOAEL = 5 mg/kg/day ( from MRID 47620509). LOAEL = 10 mg/kg/day based on decreased body weight gain and food consumption Developmental NOAEL = 10 mg/kg/day LOAEL = 60 mg/kg/day based on decreased fetal weight, and incomplete ossification of

cervical vertebrae/epiphyses/metacarpals/phalanges.Page 54 of 94 Table A.2.2 Subchronic, Chronic and Other Toxicity Profile

Guideline

No. Study Type MRID No. (year)/

Classification /Doses Results

870.3800 Reproduction and fertility effects (rat) 47620514 (2000) Acceptable/guideline 0, 80, 250, 800 ppm

M 0 : 0, 5.8, 18.0, 57.4 mg/kg/day

F 0 : 0, 6.5, 19.9, 66.2 mg/kg/day

M 1 : 0, 7.4, 23.0, 75.2 mg/kg/day

F 1 : 0, 7.8, 24.1, 78.2 mg/kg/day Parental/Systemic NOAEL = 18 mg/kg/day ♂

, 19.9 mg/kg/day ♀

. LOAEL = 57.4 mg/kg/day ♂ , 66.2 mg/kg/day ♀ based on increased liver and thyroid weights. Reproductive NOAEL = 57.4 mg/kg/day ♂

, 66.2 mg/kg/day ♀

LOAEL > HDT. Offspring NOAEL = 18 mg/kg/day ♂ , 19.9 mg/kg/day ♀

. LOAEL = 57.4 mg/kg/day ♂ , 66.2 mg/kg/day ♀ based on reduced body weight gain in both

F 1 and F 2 offspring.

870.4100b Chronic toxicity (dog) 47620505 (2000) Acceptable/guideline 0, 30, 120, 480 ppm M: 0, 1.04, 4.14, 17.29 mg/kg/day

F: 0, 1.08, 4.41, 17.32 mg/kg/day NOAEL = 17.29 mg/kg/day ♂ , 17.32 mg/kg/day ♀

. LOAEL > 17.32 mg/kg/day

870.4200 Carcinogenicity (mouse) 47638206 (2000) Acceptable/guideline 0, 60, 500, 4000/2000

ppm M: 0, 7, 63, 325 mg/kg/day

F: 0, 9, 76, 404 mg/kg/day NOAEL 63 mg/kg/day ♂ , 76 mg/kg/day ♀ LOAEL = 325 mg/kg/day ♂ , 404 mg/kg/day ♀ based on premature death (within first 20 weeks of treatment) attributed to fatty changes in liver, heart (vacuolization within myocytes) and kidney (vacuolization in cortical tubular epithelium). Hepatocellular adenomas were seen in 17/50 males in the high-dose group (vs. 6/50 control males)

870.4300 Carcinogenicity (rat) 47620511 (2001) Acceptable/guideline 0, 100, 500, 2000 ♀ /5000 ♂ ppm M: 0, 4.4, 22, 229 mg/kg/day

F: 0, 5.5, 28, 115 mg/kg/day NOAEL = 4.4 mg/kg/day ♂ , 5.5 mg/kg/day ♀ LOAEL = 22 mg/kg/day ♂ , 28 mg/kg/day ♀ based on decreased body weight gain; increased thyroid/parathyroid weight, increased liver weight and centrilobular hepatocytic hypertrophy in females; hyaline droplets and pigmented cortical tubules and cortical hemorrhagic degeneration of adrenals in males. Thyroid follicular cell tumors were observed in males of the high-dose group.

870.5100 Bacterial Reverse

Mutation 47620515 (2000) NF-149, TGAI has no reverse mutagenic potential at the limit dose under the experimental conditions.Page 55 of 94 Table A.2.2 Subchronic, Chronic and Other Toxicity Profile

Guideline

No. Study Type MRID No. (year)/

Classification /Doses Results

870.5100 Bacterial Reverse

Mutation 47620618 (2000) 149-( E )-FB (NF-149 isomer) has no reverse mutagenic potential at the limit dose under the experimental conditions.

870.5300 In vitro

Mammalian Gene

Mutation 47620516 (1997) No statistically significant increases in mutant frequency were observed following treatment with NF-149 ( ≤ 200 μ g/mL).

870.5375 In vitro

Mammalian

Chromosome

Aberration 47620517 (2000) NF-149 ( ≤ 250 μ g/mL ±S9 for 3 hours; ≤ 31.3

μ g/mL –S9 for 21 hours) showed no evidence of clastogenic or polyploidy-inducing activity in this cytogenic test system.

870.5395 Mammalian Erythrocyte

Micronucleus 47620518 (2000) NF-149 ( ≤ 2000 mg/kg) did not show any evidence of causing chromosome damage or bone marrow cell toxicity when administered orally by intragastric gavage under the experimental conditions described.

870.5550 Unscheduled DNA Synthesis in

Mammalian

Cultured Cells 47620519 (1997) NF-149 ( ≤ 2000 mg/kg) did not show any evidence of causing DNA damage in rat hepatocytes.

870.6200a Acute neurotoxicity screening battery 47620501 (2008)

Waiver request The requested waiver was denied.

870.6200b Subchronic neurotoxicity screening battery 47638205 (2001) Acceptable/guideline 0, 200, 1000, 5000 ppm M: 0, 18, 88, 453 mg/kg/day

F: 0, 21, 98, 572 mg/kg/day NOAEL = 88 mg/kg/day ♂ , 98 mg/kg/day ♀ LOAEL = 453 mg/kg/day ♂ , 572 mg/kg/day ♀ based on decreased body weight gain and food efficiency in both sexes.Page 56 of 94 Table A.2.2 Subchronic, Chronic and Other Toxicity Profile

Guideline

No. Study Type MRID No. (year)/

Classification /Doses Results

870.7485 Metabolism and pharmacokinetics (rat) 47620520 (2000) ADME after administration of [fluorinated phenyl-U- 14 C]NF-149, 10 mg/kg or 200 mg/kg.

Greater that 85% of th e administered dose (AD) was excreted within 48 hours. Urinary excretion was greater in males than females after a single dose and absorption was greater in females. The majority of radioactivity was

recovered in the feces and bile. The elimination half-lives were in the range of 14 – 34 hours for plasma and greater than 70 hours for blood. Tissue half-lives were generally less than twice those of plasma (after single dose). The highest tissue concentrations of radioactivity after 168 hrs were detected in the liver, kidney and fat. Overall tissue
accumulation was low with < 1% AD retained at 168 hr post-single dose. Excepting 149-F1, primarily a urinary metabolite (15.4% AD), major metabolites were recovered, primarily in the feces, in the following amounts in males: 149-F-3-OH-4-OH-B, 10.9%; 149-F, 4.4%; NF-149 (parent), 3.7%; 149-F-4-OH-B, 1.9%; 149-F-3-OH-B, 1.6%. The major metabolites were recovered primarily in the feces in the following amounts in females: 149-F-3-OH-4- OH-B, 27.3%; 149-F-4-OH-B, 6.2%; NF-149 (parent), 4.6%; 149-F, 2.8%; 149-F1 was

recovered as 9.9% AD in the urine of females.

870.7485 Metabolism and pharmacokinetics

(rat) 47620521 (2002) ADME of [cyclopropyl-2, 3- 14 C]NF-149, 10 mg/kg. Greater that 85% AD was excreted within 72 hours. A higher proportion of the

dose was excreted in the urine of male rats (37.7 % AD vs. 19.7% AD in females) while fecal excretion was higher in females (69.5% AD vs. 56.9% in males). The highest tissue concentrations of radioactivity after 168 hrs were detected (in order of greatest concentration) in fat, thyroid, epididymis, adrenals, liver, ovary, pancreas, prostate and skin with hair. The major metabolite detected

in urine of both males and females was CPCAGly. The major metabolites in feces were 149- F-3-OH-4-OH-B (33.4% AD ♂ , 41.4% AD ♀ ), 149-F-3-OH-B (3.2% AD ♂ , 4.5% AD ♀ ), 149-F-4-OH-B (1.6% AD ♂ , 9.8% AD ♀ ).Page 57 of 94 Table A.2.2 Subchronic, Chronic and Other Toxicity Profile

Guideline

No. Study Type MRID No. (year)/

Classification /Doses Results

870.7485 Metabolism and pharmacokinetics

(rat) 47620522 (2001) Pharmacokinetics of [cyclopropyl-2, 3- 14 C]NF149, 10 mg/kg. The following pharmacokinetic parameters were observed in plasma: Cmax =

1.3 μ g/g ♂ , 0.9 μ g/g ♀ at 2 hours (Tmax); t 1⁄2 = 6.5 hr ♂ , 7.9 hr ♀ ; AUC = 12.3 μ g·h/g ♂ , 10

μ g·h/g ♀ . The following pharmacokinetic parameters were observed in blood: Cmax =

0.31 μ g/g ♂ , 0.35 μ g/g ♀ at 2 hours (Tmax);

t 1⁄2 = 25.3 hr ♂ , 9.0 hr ♀ ; AUC = 9.88 μ g·h/g ♂

, 4.64 μ g·h/g ♀ . Absorption and elimination rates were rapid for male and female rats and similar to absorption and elimination rates observed with [fluorinated phenyl-U- 14 C]NF149.

870.7800 Immunotoxicity (mouse) 47620523 (2008)
Acceptable/guideline 0, 400, 1600, 7000 ppm M: 0, 50, 298, 1916 mg/kg/day

F: 0, 63, 388, 2542 mg/kg/day NOAEL = 1916 mg/kg/day ♂ , 2542 mg/kg/day ♀

. LOAEL > 1916 mg/kg/day ♂ , 2542 mg/kg/day ♀

. There were no statistically significant changes

in either the number of PFC/10 6 splenocytes or PFC/spleen for any treat ment group compared

to control.

870.7800 Immunotoxicity (rat) 47620524 (2008)
Acceptable/guideline 0, 300, 1800, 10800 ppm M: 0, 24, 150, 942 mg/kg/day

F: 0, 25, 150, 927 mg/kg/day NOAEL = 942 mg/kg/day, males; 150 mg/kg/day, females.

LOAEL > 942 mg/kg/day, males; LOAEL = 927 mg/kg/day, based on depression of the corpus mucosa and a decrease in spleen cells and PFC/spleen in females.Page 58 of 94 A.3 Hazard Identification and Endpoint Selection

A.3.1 Acute Reference Dose (aRfD) - Female s age 13-49 and the General Population

An acute reference dose was not defined; no effect s attributable to a sing le dose were identified in the toxicology database.

A.3.2 Chronic Reference Dose (cRfD)

Study Selected: Combined Chronic Toxicity/ Ca rcinogenicity Study in Rats

MRID No.: 47620511 Executive Summary : See Appendix A, Guideline 870.4300 Dose and Endpoint for Risk Assessment: NOAEL = 4.4 mg/kg/day based on decreased body weight gain and liver effects se en at LOAEL = 22 mg/kg/day. Comments about Study/Endpoint/Uncertainty Factors: This study is the most appropriate exposure scenario based on duration. The critical e ffect is liver toxicity which is seen across all species tested via oral admini stration. When LOAELs are co mpared among rat, dog and mouse chronic feeding studies, the rat is the most sensitive species.

A.3.3 Incidental Oral Exposure (Short- and Intermediate-Term)

Study Selected: Prenatal Developmental Study in Rabbits

MRID No.: 47620508, 47620509 Executive Summary : See Appendix A, Guideline 870.3700 Dose and Endpoint for Risk Assessment: Maternal NOAEL = 5 mg/kg/day based on decreased body weight, body weight gain a nd food consumption seen at LOAEL = 10 mg/kg/day. Comments about Study/Endpoint/Uncertainty Factors: The duration of dosing, gestation day (GD) 6-28, is appropriate for the short- term exposure scenario and protective for intermediate-term exposure (see discussion below, A3.5). A maternal LOAEL of 10 mg/kg/day, but no NOAEL, was established in the guideli ne study. A subsequent bridging study was conducted with 0, 5 or 10 mg/kg/day. No adve rse findings were observed in either treatment group. Therefore, a NOAEL of 5 mg/kg/day can be defined. Though an acceptable/guideline multigeneration study is available to represen t the population at risk (infants and young children), the prenatal developmental study chosen to protect for inhalation exposure (adult) is more protective and, therefore, applied to short- and intermediate-term in cidental oral (child)

also.

A.3.4 Dermal Exposure (Short-, Intermediate- and Long-Term)

No hazard was identified for the dermal route of exposure. Cyflufenamid was applied to shaved,

intact skin of 5 rats/sex/dose at dose levels up to and including the limit dose for 7 hours/day, 7 days/week for 4 weeks. No adverse treatme nt-related effects were observed on mortality, clinical signs of toxicity dermal irritation, body weight, body weight gain, food consumption,Page 59 of 94 food efficiency, ophthalmoscopic exams, hematology, clinical chemistry, urinalysis, absolute or relative organ weights, and gross or microscopi c pathology in either se x at the limit dose.

A.3.5 Inhalation Exposure

Study Selected: Prenatal Developmental Study in Rabbits

MRID No.: 47620508, 47620509 Executive Summary : See Appendix A, Guideline 870.3700 Dose and Endpoint for Risk Assessment: Maternal NOAEL = 5 mg/kg/day based on decreased body weight, body weight gain a nd food consumption seen at LOAEL = 10 mg/kg/day.

(Short-Term) Comments about Study/Endpoint/Uncertainty Factors: The duration of dosing, GD 6-28, is appropriate for this exposure scenario. A maternal LOAEL of 10 mg/kg/day, but no NOAEL was established in the guideline study. A subs equent bridging study was conducted with 0, 5 or 10 mg/kg/day. No adverse findings were observed in either tr eatment group. Therefore, a NOAEL of 5 mg/kg/day can be defined. Complete absorption is assumed.

(Intermediate-Term) Comments about Study/Endpoint/Uncertainty Factors: A maternal LOAEL of 10 mg/kg/day, but no NOAEL, was established in th e guideline study. A subsequent bridging study was conducted with 0, 5 or 10 mg/kg/day. No adve rse findings were observed in either treatment group. Therefore, a NOAEL of 5 mg/kg/day can be defined. Complete absorption is assumed. Though duration of dosing, GD 6-28, is less than a ppropriate for this exposure scenario and an acceptable/guideline multigeneration study is availabl e to represent the duration of exposure, the prenatal developmental study chosen to protect short-term inhalation is more protective and, therefore, chosen for intermedia te-term inhalation exposure also.

A.4 Executive Summaries

A.4.1 Subchronic Toxicity

870.3100 90-Day Oral Toxicity – Rat

In a subchronic oral toxicity study (MRI D 47638204), cyflufenamid (NF-149; 96.4% a.i.; Lot No. 31-6415-A0) was administered in the diet to Crj:CD (SD) rats (10/sex/dose) at doses of 0, 50, 300, 1800, or 10,800 ppm (equivalent to 0/0, 3.3/4.1, 20.1/24.7, 117.4/143.9, and 673.3/782.6 mg/kg/day for males/females) for 13 weeks.

No treatment-related effect was observed on mortality, ophthalmoscopic examination, or hematology.Page 60 of 94 General systemic toxicity was evident in 1800 ppm females. Decreased body weights were observed during Days 56-84 (decr 8-10%, p < 0.05). Decreased (p<0.01) body weight gain was noted on Day 1, as well as decreased overall (Day s 0-91) body weight gain was noted (decr 13%, NS). Decreased absolute and relative (decr 25-27%, p < 0.01) food consumption was also observed on Day 1. The large in itial decrease in food consum ption on Day 1 followed by much smaller decreases afterwards sugge sts a palatability issue. Incr eased (NS) incidences of the following macroscopic signs were observed (# aff ected/10 treated vs. 0/10 controls): decreased intraperitoneal adipose tissue in the males (3) and females (2); and decreased subcutaneous adipose tissue in the females (1).

The kidney was a target organ. At 1800 ppm and above, increas ed plasma blood urea nitrogen was observed in the females (incr 69-97%, p < 0.05). Increased (p< 0.05) concentrations of

ketone bodies were noted in the urine of the fema les. Increased right an d left relative kidney weights were observed in the males (incr 8-30%, p < 0.05) and females (incr 14-29%, p < 0.05). Slight hyaline droplets in rena l tubular epithelium were obser ved in all treated males and increased phosphorous was noted in both sexes (incr 33-48%, p < 0.05).

Additionally at 10,800 ppm, increased plasma bl ood urea nitrogen was observed in the males (incr 38%, p < 0.05). Right and left absolute and relative kidney weights in the 10,800 ppm males were increased compar ed to control (incr 9-30%, p < 0.05). Increased incidence of slight renal tubular vacuolization was noted in the 10,8 00 ppm females (6/10 treated vs. 0/10 controls,

p < 0.01). Increased serum phosphorous was noted in high-dose males (incr 33%, p < 0.05) and females (incr 48%, p < 0.05).

At 10,800 ppm, there was additional evidence of ge neral toxicity. An increased frequency of slight abdominal distention was not ed in the females. An increas ed frequency of urine soiling around the urethra was observed in the females (predominantly slight severity) and males (slight). Decreased body weights were observe d throughout treatment in both sexes. In both sexes, body weight losses were noted on Day 1; weekly cumulative interval body weight gains were decreased throughout treatment in both se xes. Decreased overall (Days 0-91) body weight gains were noted in both sexes. Absolute and relative food consumption were markedly decreased on Day 1 in both sexes. After Day 1, decreases ( p < 0.05) in absolute food consumption were noted on Days 7, 28, 35, 42, and 63 in males and on Days 7 through 21 in females. Relative food consumption (g/kg bw/day) was decreased ( p < 0.01) in both sexes on Day 7. The large initial decr ease in food consumption on Day 1 followed by much smaller decreases afterwards suggests a palatability issue. On Day 1, decreased (p <0.01) food efficiency was also noted in both sexes. Increased (p<0.05) incide nces of the following macroscopic signs were observed (# affected /10 treated vs. 0/10 controls): decreased intraperitoneal adipose tissue in the males (4) and females (8); and decreased subcutaneous adipose tissue in females (6).

At 10,800 ppm, the liver was also a target organ. The following increases were observed in the clinical chemistry parameters in the treated group compared to the controls at Week 14: cholesterol in both sexes, aspartat e aminotransferase in males, and γ -glutamyl transferase in bothPage 61 of 94 sexes. Increased absolute and relative to body li ver weights were noted in both sexes. Increased

incidences of evident liver lobules were observed in both sexes as well as increased incidences of slight hepatic bile duct hypertrophy and moderate hepa tic centrilobular hypertrophy.

At 10,800 ppm, the heart was a target organ. Incr eased incidence of myocarditis was observed in the males, while increased incidences of myo cardial vacuolation were noted in both sexes.

At 10,800 ppm, the thyroid was a target organ. Increased absolute and/or relative thyroid weights in both sexes were noted, and increase d incidences of slight thyroid follicular cell hypertrophy were observed in both sexes.

Increased incidence of slight Leydig cell hyperplasia was noted in the testes at 10,800 ppm.

The LOAEL is 1800 ppm (equivalent to 117.4/143.9 mg/kg/day in males/females), based on decreased body weights and body weight gains in females, decreased adipose tissue in both sexes, increased blood urea nitrogen and urin ary ketones in females, and increased relative kidney weights in both sexes. The NOA EL is 300 ppm (equivalent to 20.1/24.7 mg/kg/day in males/females).

This study is classified as acceptable/guideline and satisfies the guideline requirements (OPPTS 870.3100; OECD 408) for a subchronic oral toxicity study in the rat.

870.3100 90-Day Oral Toxicity – Mouse

In a subchronic oral toxicity study (MRID 47638202), NF-149 (Cyflufenamid; 96.4% a.i., Lot # 31- 6415-AO) was administered to 10 CD-1

mice/sex/dose in the diet at dose levels of 0, 100, 400, 1600, or 7000 ppm (equivalent to 0/0, 14.0/17.6, 50.7/70.8, 218.1/295.1, and 808.0/940.2 mg/kg/day [M /F], respectively) for up to 91 days.

No compound-related effects were noted on ophthalmoscopic examinations at any dose in either

sex. The transient abdominal distension obser ved at 7000 ppm in 3/10 males on Days 21-27 and 3-5/10 females on Days 21-32 was c onsidered to be of equivocal toxicological importance. As cecal weights (including content) in this group were not significantly increased, the cause of this finding was unknown. Additional findings of equi vocal toxicological importance at 7000 ppm

included: an increase in the number of females w ith elevated levels (15 mg/dL) of ketone bodies (7/10 treated vs. 0/10 controls) in the urine, and increases in blood ur ea nitrogen (incr 29%) and total protein (incr 13%) in the females at this dose.

At 7000 ppm, transient decreases (decr 7-8%) in mean body weights were observed in both sexes on Day 1. Additionally at this dose, the males and females displayed significant body weight losses on Day 1 (-1.87 g treated vs. 0.56 g contro ls and -1.44 g treated vs. -0.01 g controls, respectively), and body weight gain in the males re mained reduced (not statistically significant [NS]) by 65% compared to controls on Day 7. All of these findings were transient, and noPage 62 of 94 statistically significant differen ces in mean body weight and cumu lative body weight gains were observed during Days 14-91 in any treatment gr oup. In the high-dose groups, decreases were observed in both absolute (decr 40-60%) and re lative (decr 21-57%) f ood consumption during Days 1-7 and 1-14, respectively. Additionally in the females on Day 63, decreases of 29-30%

were observed in absolute and relative food consumption. Both absolute and relative food consumption were generally decreased throughout the study at this dose in both sexes; however, no further statistically significant differences we re observed. Food efficiency (%) was decreased in the males and females on Day 1 (-183.8% treate d vs. 11.8% controls an d -72.4% treated vs. - 1.7% controls, respectively). F ood efficiency remained decreased in the males at this dose on Day 7 (-253% treated vs. 6.9% controls). Beginning on Day 14 in the males and Day 7 in the females, food efficiency was generally increase d compared to controls throughout most of the remainder of the study. The Sponsor stated that the findings in body weights, body weight gain, food consumption, and food efficiency indicated a poor palatability of the dietary formulations.

The primary target organ was the liver . Clinical chemistry findings at this dose included: (i) increased ALT levels in the males (incr 515%); (ii) increased AST leve ls in the males (incr 110%); (iii) increased total cholesterol in both sexes (incr 72-91%); and (iv) decreased albumin/globulin ratio in the males (decr 29%). Both absolute and relative (to body) liver weights were increased in the males (incr 89 % each) and females (incr 112-116%). Gross

lesions were limited to dark brown liver in 5 ma les. Microscopic findings in the liver observed in all animals at the high-dose included the following: (i) incr eased incidence of centrilobular hypertrophy (slight to moderate); (ii) focal necros is (slight to moderate); and (iii) yellow pigment in the Kupffer cells (slight). A dditional hepatic findings at this dose included: slight nucleolar hypertrophy (all males) and decrea sed fat droplets (6/10 males).

Additional microscopic findings at 7000 ppm included: (i) slight myocardial vacuolation in 8 males and 2 females; (ii) testicular Leydig cel l hyperplasia (slight) in all males; and (iii) decreased secretory granules in the submandibular glands (slight to marked) in 7 males (vs. 1/10 controls).

At 1600 ppm, findings were limited to the followi ng: (i) body weight gain was transiently decreased by 91% (NS) in the males on Day 1; (ii) both absolute and relative (to body) liver weights were increased by 16-18% in the males and 21-28% in the female s; and (iii) slight centrilobular hypertrophy was noted in 4/10 mice (vs. 0 controls) of each sex. In the absence of any clinical chemistry findings, the increased liv er weights and slight centrilobular hypertrophy were considered to be adaptive and not adverse at this dose.

At 400 ppm, the minor increase in absolute liver weight (incr 19%) noted in the females was not accompanied by a significant increase in relative (to body) liver weight, a ny significant changes in clinical chemistry, or corroborat ive histopathologi cal findings.

The LOAEL is 7000 ppm (equivalent to 808.0/ 940.2 mg/kg/day [M/F]) based on increased absolute and relative liver weights, clinical chemistry (increased AST and ALT in males and increased cholesterol in both sexes, decrea sed A/G ratio in males), microscopic hepaticPage 63 of 94 lesions (centrilobular hypertrophy, focal necros is, and yellow pigment in the Kupffer cells in both sexes, and slight nucleolar hypertrophy and decreased fat droplets in males), myocardial vacuolation in both sexes, test icular Leydig cell hyperplasia, and decreased secretory granules in the submandibular gl ands in males. The NOAEL is 1600 ppm (equivalent to 218.1/295.1 mg/kg/day [M/F]).

This study is classified as acceptable/guideline and satisfies the guideline requirements (OPPTS 870.3100a; OECD 408) for a subchronic oral toxicity study in the mouse.

870.3150 90-Day Oral Toxicity – Dog

In a subchronic oral toxicity study (MRI D 47620504), NF-149 (Cyflufenamid; 95.2% a.i.; Lot No. T3G-1020) was administered in the diet to four beagle do gs/sex/dose group at doses of 0, 150, 500, or 1500 ppm (equivalent to 0/ 0, 6.5/7.5, 23.2/24.4, and 76.2/70.5 mg/kg/day in males/females) for at least 90 days.

No adverse, treatment-related effects were obs erved on mortality, clini cal signs of toxicity, ophthalmoscopic examinations, ur inalysis, or gross pathology.

Systemic toxicity was observed at 1500 ppm. Body weights were decreased in male dogs by 3-13% during Weeks 2-13, resulti ng in decreased overall (Weeks 0-13) body weight gains of 56%. In the females during Weeks 1-13, body we ights were decrease d by 7-13% and food consumption was decreased by 5-25%, resulting in decreased overall body weight gains of 42% and decreased overall food consumption of 16%.

The liver was a target organ, with adverse, treatm ent-related findings observed at 1500 ppm. Absolute and relative (to body) liver weights were increased by 16-36% in the females, and relative liver weights were increased by 26% in the males. Activated partial thromboplastin time was increased in the females by 18-31% at Weeks 6 and 13. Alkaline phosphatase was increased by 144-331% in the males and females on Weeks 6 and 13. Cholesterol was increased by 25% in the males on Week 13, and by 31-34% in th e females on Weeks 6 and 13. The following lesions were observed in the liver, compared to 0/4 controls: (i) enlarg ed, finely vacuolated midzonal and periportal hepatocyte s in the males (1/4; moderate severity) and females (1/4; slight severity); (ii) slight occas ional finely vacuolated hepatocyte s in 1/4 males; and (iii) slight enlarged finely vacuolated subcapsular hepatocyte s in the males (1/4) and females (2/4). Oil Red O staining revealed minimal to s light periportal hepatocyte fat in the 500 ppm males and females and minimal to marked/severe periportal hepa tocyte fat in the 1500 ppm males and females (compared to 0/4 controls). Oil Red O staining al so revealed marked midzonal hepatocyte fat in 1/4 females compared to 0/4 controls.

The brain was a target organ. The following lesions were observed in th e brain at 1500 ppm, compared to 0/4 controls: (i) minimal to marked v acuolation of the neuropil in the vicinity of the thalamus-cerebrum in the males (3/4; minimal to m oderate severity) and females (4/4); (ii) slight to moderate vacuolation of the superficial layer of the cerebral cortex in the males (3/4; slightPage 64 of 94 severity) and females (4/4); (iii) minimal to marked vacuolation of the white matter of the cerebrum in the males (2/4; minimal to slight severity) and females (4 /4); and (iv) slight widespread focal vacuolation of the ne uropil of the midbrain in 1/4 females.

Slight involution/atrophy of the thymus was noted at 1500 ppm in 3/ 4 males and 2/4 females, both compared to 1/4 controls.

The reviewers disagree with the Sponsor on the a dversity of the findings in the liver at 500 ppm. Absolute and relative liver weights were increased by 22% in the females, and relative liver weights were increased by 16% in the males. Activated partial thromboplastin time was increased in the females by 27% at Week 13. Al kaline phosphatase was increased by 64% in the

females on Week 13. Cholesterol was increased by 29% in the males. Oil Red O staining revealed minimal to slight periportal hepatocy te fat in the males and females (3/4 each), compared to 0/4 controls. Due to the low magni tude and/or minimal severity of these findings, the reviewers consider these findings to be an ad aptive response of the liv er to administration of the test compound at this dose.

The LOAEL is 1500 ppm (equivalent to 76.2/70.5 mg /kg/day in males/females), based on decreased body weights, body weight gains, food consumption, and liver, brain, and thymus toxicity as described above, in both sexes. The NOAEL is 500 ppm (equivalent to 23.2/24.4 mg/kg/day in males/females).

This study is classified acceptable/guideline and satisfies the guideline requirements (OPPTS 870.3150; OECD 409) for a subchronic oral toxicity study in dogs.

870.3200 21/28-Day Dermal Toxicity – Rat

In a 28-day dermal toxicity study (MRID 47620428) , NF-149 (Cyflufenamid; 95% a.i., Batch # T3G-1020) suspended in 5% gum arabic and 0.01% Tween 80 in reverse osmosis treated water was applied (2 mL/kg) to the shaved intact skin of 5 CD rats/sex/dose at dose levels of 0, 100, 300, or 1000 mg/kg/day (limit dose) for 6- 7 hours/day, 7 days/week, for 4 weeks.

No adverse treatment-related effects were obser ved on mortality, clinical signs of toxicity, dermal irritation, body weight, body weight gain, food consumption, food efficiency, ophthalmoscopic exams, hematology, clinical chem istry, urinalysis, absolute or relative organ weights, and gross or microsc opic pathology in either sex.

The LOAEL was not observed. The NO AEL is 1000 mg/kg/day (limit dose).

This study is classified as acceptable/guideline and satisfies the guideline requirement for a 28- day dermal toxicity study (O PPTS 870.3200; OECD 410) in rats.Page 65 of 94 A.4.2 Prenatal Developmental Toxicity

870.3700a Prenatal Developmental Toxicity Study – Rat

In a developmental toxicity study (MRID 47620506), Cyflufenamid (95.2%; Lot # T3G-1020) in aqueous 5% gum arabic and 0.01% Tween 80 was administered via daily oral gavage in a dose volume of 10 mL/kg to 22 time-mated presumed pregnant Sprague-Dawley rats/dose group at doses of 0, 100, 300, or 1000 mg/kg/day from gesta tion days (GD) 6-19. On GD 20, all maternal rats were euthanized, a gross necropsy performed, and the liver was weighed. Each dam’s uterus

was removed via cesarean section and its contents examined. The fetuses were examined for external, visceral, and skeletal malformations and variations.

All maternal animals survived until scheduled te rmination. Maternal bo dy weights, body weight gains, and food consumption of the treated grou ps were comparable to controls throughout the study.

At 1000 mg/kg/day, brown st aining was observed on the head in 11 dams compared to 1 control. Additionally at 1000 mg/kg/day, brown staini ng was observed on the dorsal and ventral body surfaces, sacral region, forepaws, and pinna, in 1- 5 dams per finding compared to 0 controls. The following additional findings were observed at this dose (compared to 0 controls): ungroomed coat in 3 dams; piloerection in 2 dams ; and hair loss on the forelimb in one dam. These findings indicate a general unkempt app earance and are considered treatment-related.

The following findings were noted in the 1000 mg/kg/day group at gros s necropsy, confirming the clinical signs observed dur ing the treatment period (# a ffected vs. 0 controls, unless otherwise noted): (i) brown staining on the head (9 vs. 1), limbs (8), and body surface (7); (ii) ungroomed coat (1); and (iii) hair loss on the limbs (1). Aside from these findings, the only macroscopic finding that could be attributed to tr eatment was slightly enlarged liver lobes in a single dam at 1000 mg/kg/day.

Absolute and relative (to body weight) liver weig hts were increased (p<0.01) by 12-14% at 300 mg/kg/day and by 21-22% at 1000 mg/kg/day compared to controls. Relative liver weight was also increased (p<0.01) at 100 mg/kg/day; however, this increase was minor (5%) and not

considered adverse. The reviewers consid er the increased liver weights at 300 and 1000 mg/kg/day to be treatment-related. However, because clinical chemistry and histopathology parameters are not examined in a developmental toxicity study, it is no t possible to determine whether these increases are an adap tive response or if they are adve rse. Therefore, these findings are considered equivocal.

The maternal LOAEL is 1000 mg/kg/day based on increased incidences of clinical and macroscopic observations indicating an unk empt coat appearance (e.g., brown staining, piloerection, and hair loss). The maternal NOAEL is 300 mg/kg/day.Page 66 of 94 There were no abortions, premature deliveries, complete litter re sorptions, or dead fetuses.

There were no effects of treatment on the numbers of litters, live fetuses, early resorptions, or late resorptions. Furthermore, sex ratio and post-implantation losses in the treated groups were comparable to controls.

There was no evidence of altered growth or development. Fetal body weights and placental weights in the treated groups were comparable to controls. Ther e were no dose-related effects on

ossification of the skeleton.

There were no treatment-related external, viscer al, or skeletal variations or malformations.

The developmental LOAEL was not observ ed. The developmental NOAEL is 1000 mg/kg/day (limit dose).

This study is classified acceptable/guideline and satisfies the guideline requirement (OPPTS 870.3700a; OECD 414) for a developmen tal toxicity study in rats.

870.3700b Prenatal Developmenta l Toxicity Study - Rabbit

In a developmental toxicity study (MRID 47620508) , Cyflufenamid (95.2%; Lot # T3G-1020) in aqueous 5% gum arabic and 0.01% Tween 80 was administered via daily oral gavage in a dose volume of 5 mL/kg to 26 time-mated presumed pregnant New Zealand White rabbits/dose group at doses of 0, 10, 60, or 300 mg/kg/day from gest ation days (GD) 6-28. On GD 29, all maternal rabbits were euthanized; each dam’s uterus was removed via cesarean section and its contents

examined. The fetuses were examined for exte rnal, visceral, and skel etal malformations and

variations.

At 300 mg/kg/day, seven dams abor ted prior to GD 29. Clinical signs for these animals included weight loss, little diet eaten/f ew feces, and loose feces. At 60 mg/kg/day, complete litter resorption was noted in one dam. All other maternal animals survived until scheduled termination. Hair loss on the ventral body surface was dose dependently increased over controls at 10 mg/kg/day and above (24-56% dams) compared to controls (16% dams). There were no treatment-related gross findings among the animals that survived until scheduled termination.

Maternal body weight gains in the treated gr oups at 10 mg/kg/day and above for GD 6-18 were dose-dependently decreased (p<0.01) by 44-94% co mpared to controls, resulting in decreases (p<0.05) of 28-67% compared to controls for the overall (GD 6-29) treatment period. Gravid uterine weights were unaffected by treatment. Thus, overall body weight gains adjusted for gravid uterine weights were decreased by 107-171% in the treated groups compared to controls, reflecting the decreases in body wei ght gains noted during treatment.

Maternal food consumption was decreased (p<0 .05, unless otherwise noted): by 21-26% at 10 mg/kg/day from GD 13-16; by 8-34% at 60 mg /kg/day from GD 13-21 (Not statistically significant; NS) on GD 20; and by 24 -58% at 300 mg/kg/day from GD 6-9 and GD 13-25. TotalPage 67 of 94 food consumption was 10%, 13%, and 27% lower th an controls in the 10, 60, and 300 mg/kg/day dams, respectively.

Because general toxicity was obs erved at the lowest dose tested (10 mg/kg/day), a subsequent study (MRID 47620509) was conducted at doses of 0, 5, and 10 mg/kg/day to establish a NOAEL. This study was concurrently reviewed, and a summary is included in Appendix 3 of this DER. Although no effects of treatment we re observed in the follow-up study at 5 or 10 mg/kg/day, the reviewers consid er the significantly decreased body weights, body weight gains, and food consumption in the definitive study to be strong evidence of maternal toxicity. Thus, considering the studies in conjunction, the LOAEL is 10 mg/kg/day, and the NOAEL is 5 mg/kg/day.

The maternal LOAEL is 10 mg/kg/day based on decreased body weights, body weight gains, and food consumption. The maternal NOAEL is 5 mg/kg/day.

At 300 mg/kg/day, treatment-related abortions o ccurred in 7 females, resulting in a lower number of litters and live fetuses compared to controls. Furthermore, the number of late resorptions was higher at this dose, resulting in a higher number of total resorptions. However, on a per dam basis for the animals surviving until scheduled termination, ther e were no effects of treatment on the numbers of live fetuses, early re sorptions, or late resorptions. Therefore, postimplantation losses in the treated groups were similar to controls. Additionally at 300 mg/kg/day, 17 placentae from 4 litters were pale compared to 0 controls; 10 of these placentae were found in one litter, which was observed with green amniotic fluid.

Fetal body weights were decrease d by 8% (p<0.05) in the 60 mg/kg/day females and by 13-20% (p<0.01) in the 300 mg/kg/day males and females. Skeletal retardation was evident by the following findings: (i) unossified ep iphysis at 10 mg/kg/day (5 fe tuses; 5 litters), 60 mg/kg/day (8 fetuses; 6 litters), and 300 mg/ kg/day (22 fetuses; 9 li tters); (ii) incomplete ossification and/or unossification of the metacarpals and/or phalange s at 10 mg/kg/day (3 fetuses; 2 litters), 60 mg/kg/day (6 fetuses; 4 litters), and 300 mg/ kg/day (6 fetuses; 5 lit ters); (iii) enlarged anterior/posterior fontanelle at 60 (1 fetus) and 300 (4 fetuse s; 3 litters) mg/kg/day; (iv) unossified area on the cranium at 300 mg/kg/day (2 fe tuses; 2 litters); (v) incomplete ossification of caudal vertebral element at 300 mg/kg/day (1 fetus); (vi) unossified pubes at 300 mg/kg/day (1 fetus); and (vii) unossified astragalus at 300 mg /kg/day (1 fetus). It should be noted that the investigators listed the unossification and/or in complete ossification of epiphyses, metacarpals, and/or phalanges at 60 or 300 mg/kg/day to be due to treatment, but did not mention the increased incidences in these findings at 10 mg /kg/day compared to controls. The reviewers consider the findings at 10 mg/ kg/day to be due to treatment.

There were no treatment-related external, visceral, or skeletal malformations or variations. Thus, there was no evidence of teratogenicity.Page 68 of 94 The developmental LOAEL is 60 mg/kg/day based on incidences of unossified epiphysis, unossification/incomplete ossification of th e metacarpals and phalanges, and decreased fetal body weight. The NOAEL is 10 mg/kg/day.

This study is classified acceptable/guideline and, in conjunction with the subsequent developmental toxicity study conducted to es tablish the NOAEL, satisfies the guideline requirement (OPPTS 870.3700b; OECD 414) for a developmental toxicity study in rabbits.

A.4.3 Reproductive Toxicity

870.3800 Reproduction and Fertility Effects – Rat

In a two-generation reproduction toxicity study (MRID 47620514), NF-149 (Cyflufenamid; 95.2%; Batch #T3G-1020) was administered continuous ly in the diet at di etary levels of 0, 80, 250, or 800 ppm (equivalent to 6.6/7.2, 20.5/22.0, and 66.3/72.2 mg/ kg/day in males/females, respectively) for two consecutive generations. The P generation animals (32/sex/dose group) were fed the test diets for 10 weeks prior to mating to produce the F1 litters. F1 offspring selected to be parents of the next generation (2 8/sex/dose group) were fed the same test diet concentrations as their parents. F1 parents were fed the test diets for approximately 10-11 weeks prior to mating to produce the F2 generation. Th e F2 offspring were terminated after weaning.

At 800 ppm in the P generation, four animals were euthanized prior to scheduled termination following clinical signs indicating poor general condition. One female was killed for humane reasons on GD 15, with no remarkable findings at macroscopic examination. One female was

killed in extremis on GD 9; examination of this animal at necropsy revealed a perforation in the stomach wall and associated adhesions, marked edema, inflammation, and necrosis of the submucosa, and marked hyperkeratosis and acant hosis of the keratinized region. Two other 800 ppm females were killed in extremis on LD 23. Although the condition of one of these dams was due to a mammary mass/infecti on, the second female had numerous poorly defined pale areas on the heart, confirmed microscopicall y to indicate slight acute myocar ditis of the auricle and slight chronic myocarditis with moderate myocardial mineralization of th e ventricle. Additionally in this animal, a thickened stomach wall with puncta te dark foci on the glandular mucosa was found at necropsy, with slight submucosal edema and slight necrosis/hemorrhage of the glandular region confirmed microscopically.

In the F1 generation at 800 ppm, two females were killed in extremis on LDs 1 and 2 following clinical signs indicating poor c ondition. Macroscopic examination at necropsy revealed pale liver and pale areas on the keratinized mucosa of the stomach for one female and dark areas on the mucosal surface for the other female. Microscopic examination of the stomach revealed hyperkeratosis and acanthosis, ulcers and submuc osal edema of the keratinized mucosa for one female and hemorrhage of the mucosal surface for the second female. Although the Sponsor stated that these deaths could not, with any confid ence, be attributed to treatment, the reviewers consider the deaths at 800 ppm (with the excep tion of the P dam with the mammary mass) as possibly due to treatment. The macroscopic a nd microscopic findings (p erforations, adhesions,Page 69 of 94 edema, necrosis, hemorrhage, hyperkeratosis, aca nthosis, and ulcers) may be indicative of a

direct effect of the test material on the stomach.

In addition to the findings in the stomach, lesions in the heart in one of the 800 ppm P females

killed in extremis on LD 23 were likely due to treatmen t. Microscopic examination of the numerous poorly defined pale areas on the heart noted at ne cropsy revealed slight acute myocarditis of the auricle and slight chr onic myocarditis with moderate myocardial

mineralization of the ventricle. In concurren tly submitted chronic/oncogenicity studies in rats (MRID 47620511) and mice (47638206), heart lesi ons (including chronic myocardial vacuolation in rats and mice, m yocarditis in rats, and fat depos ition in mice), were noted at the highest doses administered (2000 and 4000 ppm) . Additionally, a concurrently submitted mechanistic study (MRID 47620608) demonstrated that the test material could inhibit rat and mouse CPT activity in vitro and may contribute to cardiotoxicity.

There were no effects of treatment on body wei ghts, body weight gains, food consumption, or food efficiency in either sex in either genera tion during pre-mating. Fu rthermore, body weights, body weight gains, and food consumption in the treated dams were comparable to controls during gestation and lacta tion in both generations.

In the P generation, absolute and relative liver weights were increased by 8% (p<0.05) over controls in the 800 ppm females. None of the 800 ppm males had increased glycogen in the liver (0/32 rats) compared to 7/32 contro ls; this difference was statistica lly significant (p<0.05). In the P females, the incidence of eosinophilic foci of hepatocellular alteration was increased (p<0.05) at 800 ppm (4/26 rats) compared to 0/32 controls , and congestion was noted in the liver in 6/26 rats at 800 ppm compared to 2/32 controls.

At 800 ppm in the P generation, absolute and rela tive thyroid/parathyroid weights were increased by 13-17% over controls in males and females. These increases were significant (p<0.05), with the exception of the absolute thyroi d/parathyroid weight in P males.

In the F1 generation, an increased incidence (not significant) of increas ed vacuolation of the

zona fasiculata of the adrenal cortex was obs erved in the 800 ppm males (4/25) compared to controls (0/26).

The LOAEL for parental toxicity is 800 ppm (equivalent to 66.3/72.2 mg/kg/day in males/females, respectively) based on: mortal ity and associated myocarditis in one P dam and liver effects in the P generation. Th e NOAEL is 250 ppm (equivalent to 20.5/22.0 mg/kg/day in males/females, respectively).

There were no treatment-related e ffects on: clinical signs; birth or live birth indices; or pup sex ratio for either generation. Furthermore, when dams with complete litter loss were excluded from the data (because the mortalities were consid ered unrelated to treatment, it was apparent that the viability and lactation indices in these groups were co mparable to controls, indicatingPage 70 of 94 that offspring survival was unaffected by tr eatment. There were no treatment-related microscopic findings in the F1 or F2 pups.

At 800 ppm, pup body weight gains were decrease d by 14-23% (p<0.01) in the: F1 males and females for PND 14-21; F2 males for PND 4-7, 14 -21, and 1-21; and in the F2 females for PND 4-7 and 14-21. Body weights on PND 21 were significantly decreased ( ↓ 13%; p<0.01) in the F2 males, with non-significant decreases ( ↓ 8-9%) noted on PND 21 in the F1 males and females and

in the F2 females.

At 800 ppm, relative liver weights were incr eased by 6-10% (p<0.05) in the F1 males and females and in the F2 females compared to cont rols. Because there were no corroborating gross or microscopic lesions in the live r, these increases were considered to be an adaptive response to

the test material and not adverse.

The LOAEL for offspring toxicity is 800 ppm (equivalent to 66.3/72.2 mg/kg/day in males/females, respectively) based on decrease d pup body weights and body weight gains in both generations. The NOAEL is 250 ppm (equivalent to 20.5/22.0 mg/kg/day in males/females, respectively).

There were no effects of treatment in either ge neration on: estrous cycle; sperm parameters; mating, conception, fertility, or gestation indices; pre-coital interval; or gestation duration in either generation.

The LOAEL for reproductive toxicity was not observed. The NOAEL is 800 ppm (equivalent to 66.3/72.2 mg/kg/day in males/females, respectively).

This study is classified as acceptable/guideline and satisfies the guideline requirements (OPPTS 870.3800; OECD 416) for a two-generati on reproduction study in the rat.

A.4.4 Chronic Toxicity

870.4100b Chronic Toxicity – Dog

In a chronic oral toxicity study (MRID 47620505), NF-149 (Cyflu fenamid; 95.2% a.i.; Lot No. T3G-1020) was administered to four beagle dogs /sex/dose group in the diet at doses of 0, 30, 120, or 480 ppm (equivalent to 0/0, 1.04/1.08, 4.14/4.41, and 17.29/17.32 mg/kg/day in males/females) for 52 weeks.

No adverse, treatment-related effects were obs erved on mortality, clinic al signs of toxicity, neurological or ophthalmoscopic examinati ons, body weights or body weight gains, food consumption, hematology, urinalysis, or gan weights, or gross pathology.

Alkaline phosphatase was increased in th e 480 ppm group at Weeks 13, 26, and 52. The following findings were observed in the males: slight to moderate focal vacuolation of the zonaPage 71 of 94 glomerulosa of the adrenals in two 480 ppm dogs co mpared to minimal to slight severity in two controls; epithelial vacuolar degeneration of the epididymides in one 120 ppm dog (minimal) and two 480 ppm dogs (minimal to slight) compared to 0 controls; and minimal to slight parenchymal inflammatory cell foci of the liv er in two 30 ppm dogs, three 120 ppm dogs, and three 480 ppm dogs (slight severity ) compared to one control dog (m inimal severity). In the females, the following findings were noted in th e adrenals: focal vacuolation of the zona fasciculata in three 30 ppm dogs (minimal to moderate), two 120 ppm dogs (moderate to marked), and four 480 ppm dogs (slight to modera te) compared to two co ntrols (minimal to slight); and slight diffuse cortical hypertrophy in one 120 ppm dog and two 480 ppm dogs compared to 0 controls. The reviewers consider these responses treatm ent-related, but not adverse. The increased alkaline phosphatase and slight parenchymal inflammatory cell foci of the liver at 480 ppm represent a non-adverse response of the liver to the test compound, and the microscopic findings in the adrenals represent an adaptive response.

The LOAEL was not observed. The NO AEL is 480 ppm (equivalent to 17.29/17.32 mg/kg/day in males/females).

This study is classified as unacceptable/guideline/not upgradable and does not satisfy the guideline requirements (OPPTS 870.4100b; OECD 452) for a chronic oral toxicity study in dogs. The dogs were not dosed to the limit dose (1000 mg /kg/day), and could have tolerated a higher dose of the test compound.

A.4.5 Carcinogenicity

870.4200a Carcinogenicity Study - Rat

In a combined chronic toxicity/carcinogeni city study (MRID 47620511), Cyflufenamid (NF-149; 95.2% a.i.; Lot No. T3G-1020) was admi nistered in the diet to Crl:CD ® rats (60/sex/dose) for up to 2 years at doses of 0, 100, 500, 2000 (females only ), or 5000 (males only) ppm (equivalent to 0, 4.4/5.5, 22/28, 115 (females), and 229 (males) mg/kg/day). Additionally, groups of 20

rats/sex/dose were treated si milarly and sacrificed following 1 year of treatment.

No adverse, treatment-related effect wa s observed on mortality, food consumption, food conversion efficiency, ophthalmoscopic ex amination, hematology, or urinalysis.

Generalized systemic toxicity was indicated by the following findings. At 5000/2000 (M/F) ppm, increased incidences of the following si gns were noted in bot h sexes (unless noted otherwise) during the 2 year treatment period: skin reddening, thin, hunched posture, yellow stain (perigenital), flaccid muscle reaction (m ales only), and brown stain (males only). Additionally, palpation for masses determined th at there was an increase in the number of animals with swellings and in the total number of swellings in both sexes. The mean times of onset of the swellings were similar in the treat ed groups to the contro ls. After 1 year of treatment, increased yellow staining was noted in both sexes and brown staining in males.Page 72 of 94 At 5000/2000 ppm (M/F), body weights were decreased (statistical analysis not performed) in both sexes throughout treatment in the carcinogenicity phase (2 y ear treatment) and in males throughout treatment in the toxicity phase (1 year treatment). When all groups were combined (n=80), weekly body weight gains were decreased occasionally in the males up to Week 42 (decr 14-50%) and in the females up to Week 18 (dec r 9-29%). Cumulative body weight gains at 0-26, 0-52, and 0-74 were decreased by 12-13% in ma les and 3-7% in females. Overall (Weeks 0-104) body weight gain was decreased by 9% in males and 15% in females.

At 500 ppm and above, an increased incide nce of minimal to moderate chronic myocarditis (87-93% treated vs. 80% controls) was noted in females following 2 years of treatment. In the 2000 ppm females, an increased incidence of mi nimal to moderate chronic myocarditis was observed following 1 year of treatment.

At 500 ppm and above followi ng 2 years of treatment, hepatotoxicity was indicated by the increased incidence of slight to severe peripo rtal hepatocytic vacuola tion in females (52-55% treated vs. 40% controls).

At 5000/2000 (M/F) ppm, the following findings indica ted hepatotoxicity: (i) increased plasma

γ -glutamyl transferase concentrations in males throughout treatment and in females during Weeks 13, 26, and 52; (ii) increased absolute and relative to body liver we ights in both sexes after both 1 and 2 years of treat ment; and (iii) increased swolle n liver in both sexes following 2 years of treatment. Increased incidences of the following findings were noted following 1 year of treatment (% treated vs. controls; severity in treated group): (i ) periacinar hepatocytic hypertrophy in both sexes; (ii) hepatic centriacinar/panacinar hype rtrophy in males; and (iii) panacinar hepatocytic vacuolation in both sexes. Increased incidences of the following findings were noted following 2 years of treatment: centrilobular hepatocellu lar hypertrophy in both sexes; and hepatic focal tela ngiectasis in females.

At 500 ppm and above following 2 years of treatme nt, an increased incidence of minimal to

moderate thyroid parafollicular cell hyperplasia in females (22-32% treated vs. 15% controls)

was noted.

At 5000/2000 (M/F) ppm, absolute and relative to body thyroid weights were increased in males after 1 and 2 years and in females after 2 years. Increased incidences of thyroid follicular epithelia cell hypertrophy and pa rathyroid hyperplasia was noted in males following 1 year of treatment. Increased incidences of thyroid pa rafollicular cell hyperplasi a and thyroid follicular cell hyperplasia were noted in male s following 2 years of treatment.

It was stated by the Sponsor that the changes in the thyroid gland were considered secondary to the treatment-induced increased hepatic metabol ism of thyroid hormones. The reduction in plasma thyroid hormone levels disturbs negative feedback to the pituitary resulting in greater thyroid stimulating hormone rel ease and increased thyroid activit y. It was stated that thisPage 73 of 94 mechanism has been confirmed by the Sponsor in a separate study (study number and details were not provided).

At 500 ppm and above following 1 year of treatm ent, increased incidences of the following findings were observed in the male kidney (% treated vs. controls): slight to moderate cortical tubular hyaline droplets (30-80% vs. 5%) and pigmented cortical tubules (50-75% vs. 20%). Following 2 years of treatment, increased inciden ces of minimal to marked progressive (senile) nephropathy was observed in females (73-77% tr eated vs. 57% controls) following 2 years of

treatment.

Following 2 years of treatment, increased incide nces of minimal to ma rked adrenal cortical hemorrhagic degeneration were not ed in males (47-58% vs. 30%).

At 5000/2000 (M/F) ppm, increased incidences of the following findings in the female kidney were noted following 1 year of treatment: renal ba sophilic cortical tubules, renal cortical tubular cast(s), interstitial nephritis, and renal medulla ry cast(s). Following 2 years of treatment, increased incidences of renal pigmented co rtical tubules in both sexes were noted.

In the 2000 ppm females, increased incidences of pancreas acinar atrophy with chronic inflammation were observed following 1 and 2 years of treatment.

In the 2000 ppm females, increased incidences of thymus thyroglossal duct hyperplasia were noted following 1 and 2 years of treatment.

After 2 years of treatment, increas ed incidences of skin/subcutis mass(es) were observed grossly in the 5000 ppm males. Although an increased frequency of axillary lymph node regional to mass was noted in the 5000 ppm males, an increa se was also noted at 100 ppm; thus, the dosedependency of this effect was equivocal. After 1 year of treatment, an increased incidence of mandibular lymph node regional to mass was noted in the 2000 ppm females. A treatmentrelated increase in neoplasia (excluding the thyroid and mammary gland) was not observed; therefore, these findings were considered of equivocal toxi cological importance.

The LOAEL is 500 ppm (equivalent to 22/28 mg/kg/day in males/ females), based on effects on the heart, liver, and kidney as detailed above. The NOAEL is 100 ppm (equivalent to 4.4/5.5 mg/kg/day in males/females).

No increase in neoplasia was noted in the animals of the toxicity phase. Increased incidences of the following neoplastic lesions were noted in the 5000 ppm males and/or 2000 ppm females during the carcinogenicity phase (# affected/60 in trea ted group vs. controls): thyroid follicular cell adenoma in males (5 vs. 0; p<0.05) and females (2 vs. 0); and thyroid follicular cell carcinoma in males (2 vs. 0). The thyroid foll icular cell neoplasia incidences exceeded the reported background incidences in male CD ra ts for both adenoma (4-6%) and carcinoma (0- 2%). Background incidence was not provided for females.Page 74 of 94 In the males, the test for tre nd including all dose groups was sta tistically significant for benign follicular cell adenoma (p=0.034) and for benign follicular cell adenoma and malignant follicular carcinoma combined (p=0.004). In both cases, the te st was not significant when the high dietary concentration group was excluded.

The Sponsor stated that the effect on the thyroi d was due to hepatic enzyme induction, resulting in the accelerated degradation of thyroid hormones. This results in chronic stimulation of the thyroid through the hypothalmic-p ituitary-thyroid axis. This mode of action has been documented and is not considered problematic in humans because of differing hormone binding profiles and metabolic clearance ra te of the thyroid hormones.

At the doses tested, there was a treatment-related increase in tumor incidence when compared to controls. Thyroid follicular cell neoplasia was increased in the rats, but not thought to be important in humans. Dosing was considered adequate base d on general systemic toxicity and effects observed on the heart, liv er, kidney, pancreas, and thymus.

This study is classified as acceptable/guideline and satisfies the guideline requirements (OPPTS 870.4300; OECD 453) for a combined chronic toxicity/carcinogenicity study in rats.

870.4200b Carcinogenicity (feeding) – Mouse

In a carcinogenicity study (MRID 47638206), Cyfl ufenamid (NF-149; 95.2% a.i.; Lot No. T3G1020) was administered in the diet to CD1 mice (50/sex/dose) for up to 18 months at doses of 0, 60, 500, or 4000/2000 ppm (equivalent to 0, 7.1/ 9.0, 62.8/75.5, and 324.8/403.6 mg/kg bw/day in males/females). The dose for the high group was reduced from 4000 ppm to 2000 ppm from Week 20, due to mortality and clinical signs of toxicity.

No treatment-related effects were observed on hematology.

In the ventricle of the heart, increased inciden ces of myocardial fibros is were noted in the 500 ppm and above males (40%, each treated group vs. 32% controls; slight to marked severity) and females (16-20% vs. 6%; slight to moderate). Osteoarthrosis was observed in the femur (with joint) of males at 500 (76% treated vs. 68% controls) and 4000/2000 ppm (90%; p<0.05).

It was considered that 4000 ppm exceeded th e maximum tolerated dose, and the high dose concentration was reduced to 2000 ppm from Week 20. At 4000 ppm, there was increased mortality (particularly in females) , and thin build and labored respir ation were noted in females. Clinical signs observed in th ese animals prior to death included unresponsive/underactive behavior (one female recorded as unconscious), reduced body temperature, piloerection, pallor, partially-closed eyes, hunched/prostrate postu re, and breathing problem s. Decreased (not statistically evaluated) body weights were observed in the males throughout treatment and females during Weeks 26-78. During the first week of treatment, body weight gains were decreased by 71% in males and 48% in females. Overall body weight gains (Weeks 0-78) were decreased (p<0.01) in both sexes by 23-25%. De creased food consumption was observed in bothPage 75 of 94 sexes during the first week but this may have been a palatability issue. Food efficiency (Weeks 1-14) was decreased in the males and equivocally in the females. Macroscopic and microscopic examination supported a treatment-related effect in these animals. Gross findings included pale livers, livers with areas of change , pale skin, and thin build.

In the 4000/2000 ppm females, increased (p<0.05) incidences of fine fat deposition and fine

vacuolation were observed within myocytes of the ventricle.

At 4000/2000 ppm, absolute and relative to body liver weights were increased in both sexes. In the liver, increased incidences of areas of ch ange were noted during necropsy in the 4000/2000 ppm males. Increased incidences of the follo wing microscopic findings were observed: (i) generalized fat deposition in the liver in the males; (ii) fat deposition in the periportal/ centrilobular hepatocytes in females; (iii) widesp read, finely vacuolated hepatocytes in males; (iv) focal, finely vacuolated hepatocytes in ma les; and (v) loss of he patocytes and pooling of

blood in males.

In the 4000/2000 ppm females, increased incidences of fine vacuolation and fat deposition were noted in the cortical tubular epithelium of the kidney

.

The LOAEL is 4000/2000 ppm (equivalent to 325/404 mg/kg bw/day in males/females), based on reduced body weight gain, increased liver weight and histopathological findings in liver, heart and lungs. The NOAEL is 500 ppm (equivalent to 63/76 mg/kg/day in males/females).

Increased incidences of liver masses were observe d at necropsy in the 500 ppm males and above (30-46% treated vs. 18% cont rols; p<0.01 at 4000/2000 ppm). Hepatocellular adenoma was increased in the 4000/2000 ppm males (34%; p<0.01) compared to the concurrent controls (12%) and the range of historical controls (8-31%). There wa s no increase in incidence of hepatocellular carcinoma in males, and there were no incidences of hepa tocellular adenomas or

carcinomas in females.

At the doses tested, there was a treatment-related increase in hepatocellular adenoma when compared to controls. Dosing was considered adequate based on d ecreased body weights, body weight gains, and toxicity observed in the heart, liver, and kidney.

This study is classified as acceptable/guideline and satisfies the guideline requirement for a carcinogenicity study [OPPTS 870.4 200b; OECD 451] in mice.Page 76 of 94 A.4.6 Mutagenicity

Gene Mutation

Guideline 870.51000, In vitro Bacterial Gene Mutation ( Salmonella typhimurium/Escherichia coli )/ mammalian activation gene mutation assay, NF-149

MRID 47620515 acceptable/guideline NF-149 was tested up to the limit of solubility (1500 μg/plate, +/-S9). Precipitation of the test ma terial was observed at 1500 (T rial 1, +/-S9) and 5000 μg/plate (both trials, +/-S9). Evidence of cytotoxicity (thinning of background lawn) was observed at 5000 μg/plate in bot h trials in the presence and absence of

S9. There were no marked increases in the mean number of revertants/plate in any strain in either trial (+/-S9). Th e positive controls induced the appropriate response in all strains in the presen ce and absence of S9-activation. There was no evidence of induced mutant colonies over background.

Guideline 870.51000, In vitro Bacterial Gene Mutation ( Salmonella typhimurium/Escherichia coli )/ mammalian activation gene mutation assay, 149-( E )-FB, stereoisomer of NF149

MRID 47620515 acceptable/guideline 149-( E )-FB was tested up to the limit dose ( 5000 μg/plate, +/-S9) . Precipitation

of the test material was observed at 5000 μg/plate (+/-S9) in both trials; however, no evidence of cytotoxicity (thinning of background lawn and/or reduced number of revertants) was observed at any concentration in either trial in the presence or

absence of S9. There were no marked increases in the mean number of revertants/plate in any strain in either tr ial (+/-S9). The positive controls induced the appropriate response in all strains in the presence and absence of S9. There was no evidence of induced mutant colonies over background.

Guideline 870.5300, In Vitro Gene Mutation assay in mouse lymphoma cells

MRID 47620516

acceptable/guideline NF-149 was tested up to the limit of solub ility, upon addition of the test material to the cultures and following treatment incubation, a precipitate was observed at 200 μg/mL (+/-S9) in both trials. No mark ed increases in cytotoxicity or mutant frequency were observed at any concentrat ion with or without S9-activation in either trial. The positive controls induc ed the appropriate response both in the presence and absence of S9 in both trials. There was no evidence of induced mutant colonies over background in the presence or absence of S9-activation.

Cytogenetics

Guideline OPPTS 870.5375, In vitro Mammalian Cytogenetics (Chromosomal Aberration Assay in Human Peripheral Blood Lymphocytes)

MRID 47620517 acceptable/guideline NF-149 was tested up to cytotoxic concen trations (125 μg/mL and above, -S9 and 500 μg/mL and above, +S9). Precipitation of the test material was noted at 62.5 μg/mL and above both in the presence and absence of S9. No statistically significant increases in aberration frequenc ies were observed at any concentration evaluated in the presence or absence of S9 in either trial. The positive controls induced the appropriate response in the pres ence and absence of S9 in both trials.

There was no evidence of chromosome aberrations induced over background in the presence or absence of S9-activation.

Guideline 870.5395, In Vivo Mammalian Cytogenetics - Erythrocyte Micronucleus Assay in Mice

MRID 47620518 acceptable/guideline NF-149 was tested at up to the limit dos e (2000 mg/kg). No effects on mortality or body weight were observed at any dose. Clinical signs of toxicity (primarily hunched posture and piloerection) were observed in both sexes at 1000 and 2000 mg/kg. No significant decrease in polyc hromatic erythrocyte to normochromatic erythrocyte ratios (PCE:NCE) was observed, indicating that the test material was not toxic to the bone marrow. The positive control induced the appropriate

response. There was no significant increase in the frequency of micronucleated polychromatic erythrocytes in bone marrow after either

treatment time.

Other Genotoxicity

Guideline 870.5395 Unscheduled DNA Synthesis in Primary Rat Hepatocytes/Mammalian Cell Cultures

MRID 47620519 acceptable/guideline NF-149 was tested up to the limit dose (2000 mg/kg). No mortal ities or clinical signs of toxicity were obser ved at either dose at either exposure time. No marked

increases in the mean NNG counts were obs erved in the treated groups at either exposure time. The positive controls i nduced the appropriate response at both exposure times. There was no evidence that unscheduled DNA synthesis, as determined by radioactive tracer procedures [nuclear silver grain counts],

was induced.Page 77 of 94 A.4.7 Neurotoxicity

870.6200 Subchronic Neurotoxicity Screening Battery

In a subchronic neurotox icity study (MRID 47638205), NF-149 (Cyflufenamid; 95.4% a.i., Batch # T3G-1020) was administered in the diet to 10 CD rats/sex/group at dose levels of 0, 200, 1000, or 5000 ppm (equivalent to 0/ 0, 18/21, 88/98, and 453/572 mg/ kg/day [M/F], respectively) for 13 weeks. Neurobehavioral assessment (f unctional observational ba ttery [FOB] and motor activity testing) was performed using all rats at pre-dosing and Weeks 1, 2, 4, 8, and 12. At study termination, all surv iving animals were anes thetized and perfused in situ for collection of tissues for neuropathological examination. The ti ssues from the 5 lowest numbered rats/sex in the control and 5000 ppm groups were subjected to histopathological evaluation of brain and peripheral nervous system tissues. Po sitive control data were not provided.

No compound-related effects were observed in mortality, clinical signs of toxicity, FOB, motor activity, ophthalmoscopic parameters, gross path ology, brain measurements , or neuropathology.

One 5000 ppm male (No. 29) was sacrificed for humane reasons during Week 13. Prior to sacrifice, this animal displayed the following clin ical signs: piloerection, thin build, decreased fecal pellets, and swollen area on the left ventral abdomen. In the week prior to sacrifice, this animal ate significantly less food and lost weight. Gross necropsy revealed pale kidneys, a pale, swollen and enlarged liver, and a swollen and enlarged spleen. Histopathological examination revealed that this animal had a malignant lymphom a. This finding was considered unrelated to treatment, and no evidence of treatment-related carcinogenicity was observe d in the concurrently reviewed chronic oncogenicity study (MRID 47620511) . Body weights (no st atistical analyses reported) were decreased by 3-8% throughout th e study in both sexes, and body weight gains were decreased (p<0.01) by 33-54% in both sexes during Week 0-1. Sporadic decreases of 16- 64% in body weight gain were noted at several intervals throughout the treatment period, and a body weight loss was noted in the males for Week s 11-12 (-1g treated vs. 7g controls). Overall (Weeks 0-13) body weight gains were decreased by 9% in the males (NS) and by 18% (p<0.05) in the females. In the females, food consum ption was transiently decreased (no statistical analyses reported) by 9% during Weeks 1 a nd 2. However, overall (Weeks 1-13) food consumption was similar to controls at all doses in both sexes. Food efficiency was decreased (no statistical analyses reporte d) during Week 1 (19.0 treated vs . 27.1 controls and 7.0 treated vs. 13.8 controls) in males and females, respectively, and overall food efficien cy was also slightly decreased (10.9 treated vs. 12.0 controls and 5.0 treat ed vs. 6.2 controls) in males and females, respectively.

No compound-related effects were observed at 200 or 1000 mg/kg in either sex.

No neurological effects were observed at any dose in either sex.Page 78 of 94 The LOAEL is 5000 ppm (equivalent to 453/572 mg/kg/day, M/F) based on decreased body weight gain and food efficiency in both sexes. The NOAEL is 1000 ppm (equivalent to 88/98 mg/kg/day, M/F).

This study is classified as acceptable/guideline and satisfies the guideline requirement (OPPTS 870.6200b) for a subchronic neurot oxicity study in rats.

870.6300 Developmental Neurotoxicity Study

A.4.8 Metabolism

870.7485 Metabolism – Rat

In a rat metabolism study (MRIDs 47620520-47620522), [fluorinated phenyl-U- 14 C] or [cyclopropyl-2,3- 14 C]NF-149 (>=95% radiochemical purity ) was administered to SpragueDawley rats (4-16/sex/dose) as a single oral (gavage) dose at 10 or 200 mg/kg or as 14 consecutive daily doses at 10 mg/ kg. In addition, four male and female bile-cannulated SpragueDawley rats were administered a sing le oral dose of [fluorinated phenyl-U- 14 C]NF-149 at 10 or 200 mg/kg. Excreta, blood, and ti ssues were collected from the treated animals; radioactivity was quantified; and metabolites were isol ated and identified when possible.

At 10 mg/kg, 70-85% of the administered dos e (AD) was absorbed, while 41-51% AD was absorbed at 200 mg/kg. In plasma and blood, T

max occurred within 1-4 hours in each 10 mg/kg group, indicating relatively rapid absorption. At 200 mg/kg, absorp tion was slower (T

max occurred at 6-12 hours). The C

max ranged from 0.31-1.35 μg equi v/mL at 10 mg/kg to 3.64-17.7 μg equiv/mL at 200 mg/kg. The increase of C

max was approximately proportional to the increase in dose in the males, but not the females. The half-life in plasma was 14-16 hours in the 10 mg/kg [fluorinated phenyl-U- 14 C]NF-149 rats, 6-8 hours in the 10 mg/kg [cyclopropyl-2,3- 14 C] NF-149 rats, and 19 (males) and 34 (females) hours in the 200 mg/kg rats. Half-lives in the blood cells were longer than in the plasma . Thus, differences were observed in the pharmacokinetic parameters based on sex and do se (apart from a proportional difference based on the change in dose), as well as radiolabel position.

The compound concentrated in the blood cells of whole blood, as demonstrated by residue levels that were approximately 1.8 to 2.8-fold higher in blood cells than plasma at 72 hours after dosing or 9.7-16.0-fold higher at 168 hours after multiple dos ing. The highest concentrations outside of the GIT at 4 hours after dosing were detected: in the liver at 10 and 200 mg/kg; and in the abdominal fat at 200 mg/kg. The kidney also ha d relatively high concentr ations at the 10 and 200 mg/kg dose levels, and abdominal fat was relati vely high in each of the 10 mg/kg groups. At 168 hours after dosing of th e [fluorinated phenyl-U- 14 C]NF-149 treated groups, the liver still had the highest measured levels (0.481-0.664 μg e quiv/g at 10 mg/kg and 2.87-5.92 μg equiv/g at 200 mg/kg). The highest concentr ations were found in females at both doses. At 72 hours (terminal sacrifice) after dosing the [cyclopropyl-2,3- 14 C]NF-149 groups, the abdominal fat had the highest measured levels (1.82-2.30 μg equiv/g).Page 79 of 94 Elimination half-lives in the [fluorinated phenyl-U- 14 C]NF-149 treated groups were typically very long for the blood cells in the single dos e groups (30-52 hours) and in the multiple dose group (133-231 hours), with the highest concentr ations found in the females in each group. Conversely in the single dose groups, the half-lif e in plasma was 15-20 hours. The elimination half-lives in the pancr eas were typically among the lowest measured at 11-14 hours. In the multiple dose groups the half-life in plasma was 38-45 hours, and (excluding the GIT) the

elimination half-life in the abdominal fat wa s the lowest measured at 31-32 hours. The

elimination half-lives in tissues corroborate a sex-related difference in the pharmacokinetics of the test compound.

In the groups that were treated with e ither a single low dose or high dose of [ 14 C]NF-149 (each label), the total recoveries were 92-101% AD. Most of the dose wa s excreted within the first 24 hours (62-88% AD), with excretion in males being higher than females in each case. Excretion occurred primarily through the feces (57-88% AD), but substantial amounts were also isolated in the urine (10-32%). Minor quantities of residue were isolated in the cage wash and carcass. In the preliminary excretion balan ce study, the proportion of the dose excreted as volatiles was

below the limit of detection.

Males excreted slightly more than the females during the first 24 hours after dosing; and less

residues were found in the feces and more were f ound in the urine of males compared to females. In the 10 mg/kg [cyclopropyl-2,3- 14 C]NF-149 group, less radioactive re sidues were isolated in the feces and more in the whole body compared to the [fluorinated phenyl-U- 14 C]NF-149 group.

High amounts of the absorbed dose were found in the bile: 61-77% AD at 10 mg/kg and 34-43% AD at 200 mg/kg with highest conc entrations found in females. The Sponsor stated that a substantial proportion of the biliary radioactivity was finally elimin ated in the feces at 10 mg/kg, and re-absorption of the radioa ctivity excreted in the bile had likely occurred (enterohepatic circulation).

Excretion during the 24-hour period after dosing occurred primarily through the feces, but

substantial amounts were also isolated in the urin e. The excretion profile was similar for Days 1, 5, 10, and 14. Less residues were found in the feces and more were found in the urine of males compared to females during the 168 hour period fo llowing the last dose. No major difference was evident in the excretion prof iles, based on 14 consecutive doses compared to a single dose.

In the [fluorinated phenyl-U- 14 C]NF-149 groups, 149-F1 was the pr imary urinary metabolite in all groups and comparable levels were noted in each group; levels were higher in males than the females in each group. In the feces, the parent wa s present at only 4-5% to tal radioactive residue (TRR) in the animals administered a single gavage dose at 10 mg/kg, but was isolated at 42-50% TRR at 200 mg/kg and 24-30% TRR in the multiple dose group. The major metabolites in the feces of all groups were 149-F-3-OH-4-OH-B in bot h sexes (with higher levels observed in the females in each group) and 149-F-4-OH-B in the fe males. In the bile, the major metabolite was a mixture of conjugates, including 149-F- 3-methoxy-4-OH-B, 149-F- 3-OH-4-methoxy-B, and 149-F-4-OH-B. In the plasma, parent and meta bolites 149-F and 149-F1 were found in all dosePage 80 of 94 groups. In the liver, parent was present in all dose groups, but only at minor levels in males compared to females. Metabolite 149-F1 was pres ent in all dose groups in the liver, and levels were always higher in males than females. Metabolites 149-F6, 149-F, and 149-F4B were also found in the liver. In the kidney, parent was pr esent in all dose groups, but large differences were observed between sex and dosing regime. The metabolites 149-F1 and 149-F6 were present in the kidney in all dose gr oups. In the fat, parent was present in all dose groups. 149-Fα -OH-B was isolated in the fat in the 10 a nd 200 mg/kg single-dose males, but only minor amounts were noted in the other dose groups.

In the [cyclopropyl-2,3- 14 C]NF-149 groups, the only identified me tabolite detected in the urine was CPCA-Gly. In the feces, 149-F-3-OH-4-OH-B was the major metabolite in both sexes; 149-

F-4-OH-B was also isolated in females. The parent, 149-F-4-OH-B (males) and 149-F-3-OH-B

were also found in the feces at trace to 5% TRR.

Thus, differences were observed in metabolic profiles based on sex, dose, and radiolabel.

The metabolism of [fl uorinated phenyl-U- 14 C]NF-149 in the rat pr oceeded via two main pathways. In one pathway, NF-149 was hydrolyzed to 149-F and then reduced to the amidine

149-F1. The 149-F1 was then deaminated to form 149-F6. In the other main pathway, 149-F-3- OH-4-OH-B was formed by di-hydroxylation of the non-fluorinated phenyl ring and was subsequently converted to the methoxy deri vative. The intermed iate mono-hydroxylated

metabolites 149-F-3-OH-B and 149- F-4-OH-B were also presen t. Hydroxylation at the α position occurred as a minor pathway. A minor pathway proceeded via cleavage of the cyclopropylmethoxy moiety to form 149-F4B. The 149-F4B was then converted to a glucuronide conjugate. 149-F4B wa s also cyclized to the oxad iazole 149-F12. Conversion of NF-149 to the isomer 149-( E )-FB may represent a minor pathway. In the [cyclopropyl-2,3-

14 C]NF-149 groups, one or two hydroxylations of NF-149 took place, and these reactions sometimes proceeded to the formation of a 2-(cyclopropylcarbonylamino)acetic acid.

This metabolism study in the rat is classified acceptable/guideline and satisfies the guideline requirement for a metabolism study [OPPTS 870.7485, OPP 85-1] in rats.

A.4.9 Immunotoxicity

870.7800 Immunotoxicity – Rat

In an immunotoxicity study (MRID 47620524), NF-149, cyflufenamid (95.6% a.i., batch no. T3G-1020) was administered to 10 Crl:CD(SD) rats/s ex/dose in their diet at dose levels of 0, 300, 1800, or 10800 ppm (0, 24/25, 150, 942/927 mg/kg bw /day, M/F) for 28 days. Four days prior to termination, each rat received an intravenous dose of sheep red blood cells (SRBC, 2 x

10 8 cells). A positive control group, consisting of five male and five female rats, received a single intraperitoneal dose (50 mg /kg) of cyclophosphamide two days prior to termination. All animals were observed daily for evidence of ill -health or treatment-related changes. Body weight was recorded on Day 1, then twice week ly throughout the study period and at necropsy.Page 81 of 94 Cage food consumption was measured weekly. Afte r four weeks of treatment, all animals were subjected to a detailed necropsy; spleen and thymus weights a nd micropathology were recorded. The T-cell dependent antibody response (TDAR) was assessed using the hemolytic plaque forming cell (PFC) assay against SBRC.

There were no effects of treatment on survival, clinical signs or requi site organ weights. Decreased body weight gain was observed in males ( ↓ 23%, p < 0.01) and females ( ↓ 36, p < 0.01) at 10,800 ppm and hepatomegaly was obs erved in 5/10 high-dose females.

There was a statistically significan t decrease in the num bers of spleen cells and PFC/spleen in females receiving 10800 ppm compared to the negativ e control group. It is pertinent to note that

there was substantial individual va riation within concurrent cont rol values and treatment groups. However, the distribution of indi vidual data clearly suggested a decreased activity pattern in the high-dose females compared with negative cont rol and other treated groups. There were no significant changes in the numbers of PFC/10 6 cells or PFC/spleen in fe males treated with 300 or 1800 ppm and in all male treatment groups compared with the negative control group. Positive control of cyclophosamide demonstrated a near complete suppression of the antibody response to the challenge with SRBC in both male and female rats.

Under the conditions of this study, the LOAE L for T-dependent antibody response in females was 10800 ppm (equivalent to 927 mg/kg/day) and the NOAEL is 1800 ppm (equivalent to 150 mg/kg/day). The LOAEL fo r males was not established and the NOAEL is 10800 ppm (equivalent to 942 mg/kg/day).

This study is classified as acceptable/guideline and satisfies the guideline requirement for an immunotoxicity study (OPPT S 870.7800) in the rat.

870.7800 Immunotoxicity - Mouse

In an immunotoxicity study (MRID 47620523), NF-149, cyflufenamid (95.6% a.i., Batch no. T3G-1020) was administered to 10 Crl:CD1(ICR) mice/sex/dose in thei r diet at dose levels of 0, 400, 1600, or 7000 ppm (0, 50/63, 298/388, 1916/2542 mg/ kg bw/day, M/F) for 28 days. Four days prior to termination, each mouse received a single intravenous dose of sheep red blood cells (SRBC, 4 x 10 8 cells)

. A positive control group, consisting of six male and six female mice, received 20 mg cyclophosphamid/kg/day via gavage fo r 5 consecutive days (7 to 3 days prior to termination). All animals were observed for evid ence of ill-health or treatment-related changes. Body weight was recorded on Day 1, then tw ice weekly throughout th e study period and at necropsy. Cage food consumption was measured w eekly. After four weeks of treatment, all animals were subjected to a detailed necropsy; spleen and thymus we ight and micropathology were recorded. The T-cell dependent anti body response (TDAR) was assessed using the hemolytic plaque forming cell (PFC) assay against SRBC.

One female receiving 7000 ppm was euthanized on Day 23 due to poor clinical condition; hypoactivity, reduced body temperatur e, pallor and tachypnea were all observed prior to death.Page 82 of 94 Necropsy revealed a pale, enlarged liver. There were no effects of treatment on food consumption, body weight or body we ight gain, clinical signs or requisite organ weights in surviving animals. Upon macroscopic examinat ion, enlarged livers were observed in all male and the majority of female (7/10) animals tr eated with 7000 ppm and one male and one female treated with 1600 ppm. Liver weights were not recorded. Macroscopic changes were also observed in the spleens of high-dose group animals. Enlarged (4/10 vs. 1/10 control) or pale (2/10 vs. 0 control) spleens were observed at low incidence, not accompanied by weight change and only present in males. While the toxicologi cal significance is unclear, these gross findings

are not an indicator of immunosuppression.

There were no statistically significant ch anges in either the number of PFC/10 6 cells or PFC/spleen for any of the treatment groups compared to the negative control. The data suggest

that under the conditions of th is study, cyflufenamid did not significantly a lter the T-cell dependent antibody response. The positive control demonstrated marked immunosuppression. Upon challenge with SRBC PFC/10 6 cells, PFC/spleen, and cells/spl een were all suppressed by cyclophosphamide treatment.

No natural killer (NK) cell activity was evalua ted because the available toxicity database revealed no abnormal findings in organ weights and histopathology in the immune organs. By the weight of evidence, it is suggested that the immune system is not a primary target organ for cyflufenamid exposure.

Under the conditions of this study, the LOAE L for the T-cell dependent antibody response

was not established and the NOAEL is 7000 ppm (equivalent to 1916/2542 mg/kg/day, M/F).

This immunotoxicity study is classified acceptable/guideline and satisfies the guideline requirement for an immunotoxicity study (OPPTS 870.7800) in the mouse.

A.4.9 Special/Other Studies

In Vitro Inhibition of Porcine Thyroid Peroxidase

This non-guideline study (MRID 47620606) was perfor med to determine if NF-149 could inhibit porcine thyroid peroxidase, using an in vitro assay to measure activity of this enzyme. A microsomal fraction of porcine thyroid glands wa s prepared, and the inhibitory potential of NF149 (Lot No. T3G-1022; 95.9% a.i.) was determined by monitoring the oxid ation of guaiacol, a substrate for thyroid peroxidase. Concentr ations of the test compound used were 1x10 -5 M,

1x10 -4 M, 5x10 -4 M, and 1x10 -3 M; a vehicle contro l (90% DMSO solution) was also included. 6-Propyl-2-thiouracil (PTU), a known inhibitor of thyroid per oxidase, was included as a positive control, and was tested at concentrations of 1x10 -5 M, 5x10 -5 M, 1x10 -4 M, and 1x10 -3 M.

Compared to the vehicle control, no suppression of the absorbance increase was observed at any

concentration of NF-149. The limit of solubility was exceeded at 5x10 -4 M and higher, as aPage 83 of 94 white turbid solution was observed. Therefore, the test compound did not inhibit porcine thyroid peroxidase at the concentrations tested.

The positive control (PTU) dilutions caused a con centration-dependent decrease in absorbance change, indicating PTU inhibited porcine thyroid peroxidase as expecte d. Inhibition was 100% at the highest concentration (1x10 -3 M); and the calculated IC 50 was 6.6x10 -5 M.

This study is classified as acceptable/non-guideline . It provides credible supplemental information regarding the ability of the te st compound to inhibit thyroid peroxidase in vitro

.

Induction of Hepatic Drug Metabolizing Enzymes in Male Mice

This non-guideline study (MRID 47620607) was perf ormed to determine if NF-149 could induce hepatic drug metabolizin g enzymes in male mice. Groups of five male CD-1 (Crj:CD-1 [ICR]) mice/dose group were administered NF-149 (Lot No. T3G-1022; 95.9% a.i.) in the diet at concentrations of 0, 60, or 2000 ppm (equivalent to 0, 9.08, and 292.8 mg/kg/day) for 14 days. A positive control group of five male CD-1 mi ce was concurrently administered the known hepatic enzyme inducer phenobarbital in the diet at a concentration of 500 ppm (equivalent to 62.8 mg/kg/day) for 14 days. Clinical obser vations, body weights, food consumption, liver weights, and macroscopic liver observations were recorded. Mi crosomal and cytosol fractions were prepared from the livers of all mice, and cytochrome P-450 leve ls and activities of sulfotransferase and UDP-glucur onyltransferase were determined.

There were no test compound-rela ted effects of treatment on mo rtality, clinical signs, body weights, food consumption, liver we ights, or gross liver pathology.

In the 2000 ppm NF-149 group, cytochro me P-450 levels were increased (p ≤ 0.05) by 16% compared to controls; UDP-glucuronyltransferase a nd sulfotransferase were similar to controls. While the reviewers agree with the Sponsor that this increase is sugge stive of induction of hepatic phase 1 enzymes, it must be noted that th e level of induction is slight compared to the known inducer, phenobarbital (PB).

Absolute and relative (to body) liv er weights were increased (p ≤ 0.01) by 27-29% in the 500 ppm PB mice. In the 500 ppm PB group, cyto chrome P-450 levels were increased (p ≤ 0.01) by 101%, and sulfotransferase activity wa s increased by 92%. UDP-glucur onyltransferase was not affected by PB administration.

This study is classified as acceptable/non-guideline . It provides supplemental information regarding the ability of the test compound to i nduce hepatic drug metabolizing enzymes in male

mice.

In Vitro Inhibition of Rat and Mouse Cardiac Carnitine Palmitoyltransferase

It was stated that in the chronic studies on NF-149 (assumed by the reviewers to be the concurrently submitted chronic toxicity/car cinogenicity study in rats [MRID 47620511] and 18-Page 84 of 94 month carcinogenicity study in mice [MRID 476 38206]), heart lesions, including myocardial vacuolation in rats and mice, m yocarditis in rats, and fat depos ition in mice were noted at the highest doses administered. The inhibition of carnitine palmitoyltransferase (CPT) by NF-149 was suspected as a mechanism, as CPT plays an im portant role in the heart in the transport of long chain fatty acids into the mitochondrial matr ix. Therefore, this non -guideline study (MRID 47620608) was performed to determine if NF-149 could inhibit rat and mouse CPT, using an in

vitro assay to measure activity of this enzyme. Microsomal fractions of rat and mouse hearts were prepared, and the inhib itory potential of NF-149 (Lot No. T3G-1022; 95.9% a.i.) was determined by measuring the rel ease of free CoASH from palmit oyl CoA using 5,5'-dithiobis (2- nitrobenzoic acid) (DTNB) as a substrate. C oncentrations of the test compound used were 0.3, 0.5, and 1.0 mM. Amiodarone, oxfenicine, and ad riamycin, substances known to inhibit CPT activity, were also assayed as positiv e controls to validate the assay.

NF-149 inhibited rat and mouse CPT by 52% and 51%, respectively, at a concentration of 1.0 mM. Slight inhibition was also observed at 0.5 mM (15% and 16% in rat and mouse, respectively), while no inhibition wa s noted at 0.3 mM in either sp ecies. Each positive control substance showed dose-dependent inhibition of CP T in both species. Additionally, it was stated that the positive controls inhibited CPT at si milar concentrations to those reported in the literature, validating the accuracy of the assay.

Fatty acids are the primary oxidative fuel for cardiac energy production, with lactate and glucose providing additional sources of energy. As CPT is involved in the transport of long chain fatty acids into the mitochondrial matrix, inhibition of CPT could result in depression of myocardial metabolism, including reductions in fatty acid oxidation, TCA cycle, and mitochondrial respiration. Therefore, it is possible to attribute the myocar dial lesions observed in the concurrently submitted chronic toxicity/carcinogenic ity studies to inhibition of CPT by the test compound.

This study is classified as acceptable/non-guideline . It provides credible supplemental information regarding the ability of the test compound to inhibit rat and mouse CPT in vitro

.Page 85 of 94 Appendix B. Metabolism Summary Table

Table B. Tabular Summary of Metabolites and Degradates.

Chemical Name (other names in parentheses)

and Structure Matrix Percent TRR (PPM) 1
Matrices - Major Residue (>10%TRR) Matrices - Minor Residue (<10%TRR) Cyflufenamid (NF-149)

[ N

( Z) ]- N -[[(cyclopropylmethoxy)amino][2,3-

difluoro-6- (trifluoromethyl)pheny l]methylene]benzeneacetamid

( Z

)]- N -[ α -(cyclopropylmethoxyimino)-2,3- difluoro-6-(trifluoromethyl)benzyl]-2- phenylacetamide

F

F N H N O O

CF 3

Apple fruit and leaves; Cucumber fruit and leaves; Wheat forage, straw, husks, heads, and grain

Goat milk, muscle and fat

Water Apple (91 DAT, mature) Fruit: 66.2 (0.012)

Leaves: 16.9 (0.131)

Cucumber Fruit (L): 55.0-96.3 (0.019-0.068) Fruit (H): 85.8 (0.179) Leaves (L): 42.3-97.9 (0.542-2.91) Leaves (H): 67.5-96.3 (4.50-6.69)

Wheat (Fluorinated Phenyl Label Study) Forage: 90.16-104.07 (0.621-2.084) Straw, immature:

29.33/38.02 (0.169/0.737)
Straw, mature:

33.40/40.78 (0.223/0.958 Husk, mature:

10.33/19.55 (0.010/0.106) Head, immature: 28.95 (0.031) Grain, mature: 1.95 (<0.001)

Wheat (Cyclopropyl Label Study) Forage: 88.77-101.39 (0.629-5.064) Straw, immature:

59.06/64.2 (0.179/1.271)
Straw, mature:

55.52/65.17 (0.171/1.272) Husk, mature,:

16.89/19.92 (0.057/0.197) Head, immature:

20.09/21.74 (0.024/0.109) Grain, mature: 6.59 (<0.004) Lactating

Goat Milk: 6.5 (0.003) Fat: 79.8 (0.120) Muscle: 21.5 (0.007) Rat (feces) male,30.2%; female

23.5% AD

Water Not available 2 Not available 2Page 86 of 94 149- (E) -FB

( E )- N -( α -cyclopropyl-methoxyimino-2,3-difluoro6-trifluoromethylbenzyl)-2-phenylacetamide

CF 3 F

F N H N O O

Apple fruit and leaves; Cucumber fruit and leaves; Wheat forage, straw, husks, heads, and grain

Goat fat Apple (91 DAT, mature) Fruit: 1.33 (<0.001)
Leaves: 0.670 (<0.005)

Cucumber Fruit (L): 1.03-1.32 (<0.001-0.001) Fruit (H): 1.36 (0.003) Leaves (L): 1.25-1.79 (0.16-0.51) Leaves (H): 1.25-1.48 (0.087-0.098)

Wheat (Fluorinated Phenyl Label Study) Forage: 2.60 (0.021)

Straw, immature: 2.04/ 0.89 (0.012/0.017)
Straw, mature: 0.72 (0.17) Husk, mature: 0.34/0.35 (<0.001/0.002) Head, immature: 1.34 (0.001) Grain, mature: 0.09 (<0.001)

Wheat (Cyclopropyl Label Study) Forage: 2.25-3.03 (0.020-0.114) Straw, immature: 0.54/3.88 (0.002/0.077)
Straw, mature: 4.28/4.08 (0.013/0.080) Husk, mature: 1.68/0.38 (0.006/0.004) Head, immature: 1.05/1.17 (0.001/0.006) Lactating

Goat Fat: 2.5 (0.004)

149-F

N

-(cyclopropyl-methoxy-2,3-difluoro-6- trifluoromethylbenzamidine Wheat (Fluorinated Phenyl Label Study) Straw immature: 0.28 (0.005)

Straw mature: 2.12/4.10 (0.014/0.096)

Lactating

Goat Fat: 1.4 (0.002)

Liver: 14.5/7.4 (0.016/0.069)

Rat (feces) male, 3.1%; female,

3.5% ADPage 87 of 94 CF 3 F

F NH 2 N O

Wheat straw

Goat fat and liver

Water Water Not available 2 Not available 2

149-F1

2,3-difluoro-6-trifluoromethylbenzamidine

CF 3 F

F NH 2 NH

Goat milk, muscle, kidney and liver

Water Lactating

Goat Muscle: 45.5 (0.014) Kidney: 14.9/55.5 (0..002/0.120)

Liver: 30.6/29.6 (0.034/0.276) Milk: 61.6 (0.025) Rat (urine) male, 14.4%; female,

6.3% AD

Water Not available 2 Not available 2

149-F6

2,3-difluoro-6-trifluoromethylbenzamide

CF 3 F

F NH 2 O

Rotated carrot foliage (120-day PBI)

Goat milk, muscle, kidney and liver

Water Carrot Foliage: 32.5 (0.013) Lactating

Goat Muscle: 29.9 (0.009) Kidney: 17.2/13.9 (0.003/0.030)

Liver: 3.3/8.6 (0.004/0.080) Milk: 4.3 (0.002)

Rat (urine) male, 3.0%; female,

0.6% AD

Water Not available 2 Not available 2

149-F- α -OH-B

( Z

)- N -( α -cyclopropyl-methoxyimino-2,3-difluoro-6- trifluoromethylbenzyl)-2-hydroxy-2- phenylacetamide Cucumber Fruit (L): 0.82-7.14 (0.001-0.002)

Leaves (L): 0.44-2.17 (0.013—0.041) Leaves (H): 0.55-1.27 (0.038—0.084)

Wheat (Fluorinated Phenyl Label Study) Straw, immature: 2.69/4.64 (0.015-0.090)
Straw, mature: 3.40/4.46 (0.030-0.080)Page 88 of 94 CF 3 F

F N H N O O

OH

Cucumber fruit and leaves; Wheat straw, husks, heads, and grain

Goat fat Husk, mature: 2.36/3.07 (0.003-0.013) Head, immature: 1.34 (0.001) Grain, mature: 0.25 (<0.001)

Wheat (Cyclopropyl Label Study) Straw, immature: 1.36 (0.004) Husk, mature: 1.32 (0.013)

Lactating

Goat Fat: 1.6 (0.002)

149-F- α -OH-B-Glu

Not Provided

CH 2 OH

CF 3 O

OH OH F

F N H N O O

O
O H

Cucumber leaves Cucumber Leaves (L): 0.61-3.36 (0.015-0.042) Leaves (H): 0.12-1.20 (0.008-0.080)

149-F-2-OH-B

N

-( α -cyclopropyl-methoxyimino-2,3-difluoro-6- trifluoromethylbenzyl)-2-(2- hydroxyphenyl)acetamide

F

F N H N O O

CF 3 OH

Wheat straw, husks, heads, and grain Wheat (Fluorinated Phenyl Label Study) Straw,

immature:1.10/0.93 (0.006/0.018) Straw, mature:2.53/1.85 (0.017/0.043) Husk, mature:1.54/1.47 (0.001/0.008) Head, immature: 1.65 (0.002) Grain, mature:0.23 (<0.001)

Wheat (Cyclopropyl Label Study) Straw, immature: 1.19/0.65 (0.004/0.013)
Straw, mature: 2.33/1.21 (0.007/0.024) Husk, mature: 0.91/0.94 (0.003/0.009) Head, immature:

0.11/0.32 (<0.001/0.002)Page 89 of 94 149-F-3-OH-4-OH-B

( Z

)- N -( α -cyclopropyl-methoxyimino-2,3-difluoro-6- trifluoromethylbenzyl)-2-(3,4- dihydroxyphenyl)acetamide Rat (feces) 11-27% AD

149-F-4-OH-B

( Z

)- N -( α -cyclopropyl-methoxyimino-2,3-difluoro-6- trifluoromethylbenzyl)-2-(4- hydroxyphenyl)acetamide

F

F N H N O O

CF 3 OH

Cucumber fruit and leaves; Wheat straw, husks, heads, and grain

Goat fat Cucumber Fruit (L): 1.93-8.57 (0.002-0.006) Fruit (H): 7.59 (0.016) Leaves (L): 0.18-0.57 (0.004-0.013) Leaves (H): 0.06-0.30 (0.004-0.020)

Wheat (Fluorinated Phenyl Label Study) Straw, immature: 2.0/2.88 (0.017/0.039)
Straw, mature: 1.96/2.24 (0.015/0.046) Husk, mature: 2.21/2.67 (0.003/0.12) Head, immature: 3.45 (0.004) Grain, mature: 0.40 (<0.001)

Wheat (Cyclopropyl Label Study) Straw, immature: 1.37 (0.027)
Straw, mature: 5.12/2.04 (0.016/0.04) Husk, mature: 4.71/2.02 (0.016/0.020) Head, immature:

0.22/0.39 (<0.001/0.002) Lactating

Goat Fat: 1.5 (0.002)

Rat (feces) male, 5.3%; female,

18.5% AD

149-F-4-OH-B-Glu

N

-( α -cyclopropylmethoxyimino-2,3-difluoro-6- trifluoromethylbenzyl)-2-(4- β

- D -glucopyranosyl)- phenyl)acetamide Apple (91 DAT,mature) Leaves: 8 (0.062)

Cucumber Fruit(H): 1.42 (0.003) Leaves(L): 0.53-8.54 (0.013-0.108) Leaves(H): 0.56-3.82 (0.039-0.254)Page 90 of 94 O

OH OH O H O

F

F N H N O O

CF 3 CH 2 OH

Apple leaves; Cucumber fruit and leaves;

149-F4B-Glu

N

-( β -glucopyranosylimino-2,3-difluoro-6- trifluoromethylbenzyl)-2-phenyl-acetamide

CH 2 OH

O

OH OH O H O

F

F N H N O

CF 3

Apple leaves

Wheat straw and husks Apple Leaves: 6.6 (0.051)

Wheat (Fluorinated Phenyl Label Study) Straw, immature: 3.67/2.50 (0.021/0.049)
Straw, mature: 0.70 (0.16) Husk, mature: 3.48 (0.019)Page 91 of 94 149-F11

( Z

)]- N -( α -cyclopropylmethoxyimino-2,3-difluoro-6- (trifluoromethylbenzyl)carbamoylacetic acid

N H N O O F

F

CF 3 O

OH

Water Water Not available 2 Not available 2

Apple: 47620805; 0.241 lb ai/A (Broadcast foliar spray); 0-91-day PHI (mature). Cucumber: 47620806; 0.045 and 0.178 lb ai/A (Broadcast foli ar spray); 0-31-day PHI/7/35-day PHI (0.178 lb ai/A). Wheat: 47620809; 0.044 and 0.0178 lb ai/A (Broadcast foliar spray); 0-77-day PHI (mature). Wheat: 47638210; 0.044 and 0.0178 lb ai/A (Broadcast foliar spray); 0-53-day PHI (mature). Rotational Crops (Carrot, Lettuce and Wheat): 47620811; 0.045 lb ai/A 1x, applied to bare sandy loam soil; 30,120,270-days PBI. no parent or metabolites identified. Goat; 47620810; 1.2 and 13.3 ppm; 1x; 5 days; 23-hour after last dose administration.. Rat: MRID No 47620520 through 47620522; study design of 8 rats/sex receiving 10 mg/kg for 14 consecutive days, [fluorinated phenyl-U- 14 C]NF-149. Water: DP#360795, R.F.H. Bohaty, 3/4/2010

1 When the study was conducted at both a nominal and high applicat ion or dosing rate, both results are listed in the table; (L)

- low rate; (H) - high rate.

2 Data are not available on individual components of the residue in water.Page 92 of 94 Appendix C. Review of Human Research

This risk assessment relies in part on data from studies in which adult human subjects were intentionally exposed to a pest icide to determine their dermal and inhalation exposure. Many such studies, involving exposure to many different pesticides, comprise generic pesticide exposure databases such as PHED and the Outdoo r Residential Exposure Task Force (ORETF). EPA has reviewed all the studies supporting th ese multi-pesticide generic exposure databases,

and has found no clear and convinc ing evidence that the conduct of any of them was either fundamentally unethical or signifi cantly deficient relativ e to the ethical sta ndards prevailing at the time the research was conducted. All appl icable requirements of EPA’s Rule for the Protection of Human Subject s of Research (40 CFR Part 26) ha ve been satisfied, and there is no regulatory barrier to conti nued reliance on these studies.Page 93 of 94 Appendix D. International Residue Limit Status Sheet



### INTERNATIONAL RESID UE LIMIT STATUS

Chemical Name: [ N ( Z

)]- N

- [[(cyclopropylmethoxy)- amino][2,3-difluoro-6- (trifluoromethyl)phenyl]- methylene]benzeneacetamid

e Common Name: Cyflufenamid □ Proposed tolerance

□ Reevaluated tolerance

X Other-Recommended Date: 11/10/09

Codex Status (Maximum Residue Limits) U. S. Tolerances

X No Codex proposal step 6 or above

□ No Codex proposal step 6 or above for the crops requested Petition Number: PP#8F7488

DP#: 364556

Other Identifier:

Residue definition (step 8/CXL): N/A Reviewer/Branch: M. Negussie/RAB3

Residue definition: cyflufenamid

Crop (s) MRL (mg/kg) Crops Proposed Tolerances (ppm)

Fruit, pome, group 11 0.06

Apple, wet pomace 0.10

Vegetable, cucurbit, group 9 0.07

Fruit, small vine climbing, subgroup 13-07F, except fuzzy

kiwifruit 0.15

Grape, raisin 0.30

Berry, low growing, subgroup 13-07G, except cranberry 0.20

Limits for Canada Limits for Mexico

X No Limits

□ No Limits for the crops requested X No Limits

□ No Limits for the crops requested

Residue definition: N/A Residue definition: N/A

Crop(s) MRL (mg/kg) Crop(s) MRL (mg/kg)

Notes/Special Instructions: S. Funk, 11/10/2009.Page 94 of 94 Appendix E. Data Required as a Condition of Registration

Table E. Description of Outstanding Studies

Guideline Number: 870.6200 Study Title : Neurotoxicity Battery (Acute Studies) Rationale for Requiring the Data

This is a new data requirement under 40 CFR Pa rt 158 as a part of the data requirements for registration of a pestic ide (food and non-food uses).

The Neurotoxicity Screening Battery (OPPT S 870.6200) is designed to evaluate the potential adverse effects on the nervous syst em from exposure to pe sticide chemicals. The Agency believes that the guideline studi es are inadequate in their assessment of

behavioral effects and do not us e optimal methods to evaluate the potential toxicity to the

nervous tissue structure and function. To de tect and characterize these potential effects more fully, a battery of more sensitive testing is required. Th e objective of this neurotoxicity battery testing is to evaluate the incidence a nd severity of the functional and/or behavioral effects, the level of motor activity, and the histopathology of the nervous system. The acute neurotoxicity st udy is required to detect possible effects resulting from a single exposure.

Practical Utility of the Data

How will the data be used?

The acute neurotoxicity study provides critic al scientific information needed to characterize potential hazar d to the human population on the nervous system from pesticide exposure. This study can provide da ta on a wide range of functional tests for evaluating neurotoxicity includ ing sensory effects, neuromus cular effects, learning and memory and histopathology of the nervous syst em For example, animal studies with organophosphorous chemicals have shown neur otoxicity to be the primary toxic endpoints (e.g., cholinesterase inhibition) of concerns in rodents and non-rodents. This animal study can be used to select endpoints and doses for use in risk assessment of all exposure scenarios and is considered a primar y data source for reliable acute reference dose calculation. The Agency has established an oral acute reference dose (RfD) for assessing dietary risks for a number of chemicals (e.g., organophosphates and carbamates) here neurotoxicity was the most sensitive endpoint of concern.

How could the data impact the Agency's future decision-making?

If the acute neurotoxicity study show that cy flufenamid poses a grea ter risk than that given in the interim decision’s conclusion, th e risk assessments for cyflufenamid may need to be revised to reflect the magnitude of potential risk derived from the new data.

If the Agency does not have this data, a 10X database uncertainty factor may be applied when conducting a risk assessment usi ng the currently available studies.