Page 1 of 87 UNITED STATES ENVIRONMENTAL PROTECTION AGENCY WASHINGTON, D. C. 20460 OFFICE OF PREVENTION, PESTICIDES AND TOXIC SUBSTANCES Date: 6/ 29/ 2006 MEMORANDUM:

SUBJECT: Benthiavalicarb Isopropyl: Human Health Risk Assessment for Proposed Uses on Imported Grapes and Tomatoes. PC Code: 098379, Petition No: 3E6545, DP Barcode: D330555.

Regulatory Action: Section 3 Registration Action Risk Assessment Type: Single Chemical Food Only ( Import Tolerances)

FROM: Douglas Dotson, Ph. D., Chemist Kim Harper, Toxicologist Registration Action Branch 2 Health Effects Division ( 7509C)

AND Dennis McNeilly, Chemist Karlyn Bailey, Toxicologist Michael Doherty, Ph. D., Chemist Registration Action Branch 2 Health Effects Division ( 7509C)

THROUGH: Richard Loranger, Ph. D., Branch Senior Scientist Christina Swartz, Branch Chief Registration Action Branch 2 Health Effects Division ( 7509C)

TO: Tamue Gibson/ Mary Waller, RM 21 Fungicide Branch Registration Division ( 7505C)

This document is a revised version of the 3/ 2/ 2006 risk assessment ( D322937). It has been prepared in response to comments from the Office of General Council regarding cumulative risk as well as the use of average field trial residues and percent crop imported data. There have been no changes in recommended tolerances or estimates of dietary risk. Page 2 of 87 Table of Contents 1.0 Executive Summary....................................................................................................... 4 2.0 Ingredient Profile........................................................................................................... 9 2.1 Summary of Registered/ Proposed Uses ................................................................ 9 2.2 Structure and Nomenclature................................................................................. 9 2.3 Physical and Chemical Properties ....................................................................... 10 3.0 Metabolism Assessment............................................................................................... 11 3.1 Comparative Metabolic Profile ........................................................................... 11 3.2 Nature of the Residue in Foods .......................................................................... 11 3.2.1. Description of Primary Crop Metabolism................................................ 11 3.2.2 Description of Livestock Metabolism...................................................... 12 3.2.3 Description of Rotational Crop Metabolism, including identification of major metabolites and specific routes of biotransformation...................... 12 3.3 Environmental Degradation................................................................................ 12 3.4 Toxicity Profile of Major Metabolites and Degradates ........................................ 12 3.5 Summary of Residues for Tolerance Expression and Risk Assessment ............... 13 3.5.1 Tabular Summary ................................................................................... 13 3.5.2 Rationale for Inclusion of Metabolites and Degradates............................ 13 4.0 Hazard Characterization/ Assessment.............................................................................. 14 4.1 Hazard Characterization..................................................................................... 14 4.2 FQPA Hazard Considerations............................................................................. 25 4.2.1 Adequacy of the Toxicity Data Base ....................................................... 25 4.2.2 Evidence of Neurotoxicity ...................................................................... 25 4.2.3 Developmental Toxicity Studies.............................................................. 25 4.2.4 Reproductive Toxicity Study .................................................................. 26 4.2.5 Additional Information from Literature Sources...................................... 26 4.2.6 Pre and/ or Postnatal Toxicity.................................................................. 26 <IP_ADDRESS> Determination of Susceptibility.................................................... 26 <IP_ADDRESS> Degree of Concern Analysis and Residual Uncertainties for Pre and/ or Post natal Susceptibility ................................................... 26 4.3 Recommendation for a Developmental Neurotoxicity Study ............................... 26 4.4 Hazard Identification and Toxicity Endpoint Selection........................................ 27 4.4.1 Acute Reference Dose ( aRfD)  All Populations...................................... 27 4.4.2 Chronic Reference Dose ( cRfD) ............................................................. 27 4.4.3 Non Dietary Exposure for Handlers........................................................ 27 4.4.4 Margins of Exposure .............................................................................. 27 4.4.5 Recommendation for Aggregate Exposure Risk Assessments.................. 27 4.4.6 Classification of Carcinogenic Potential .................................................. 27 4.5 Special FQPA Safety Factor ............................................................................... 27 4.6 Endocrine disruption .......................................................................................... 28 Page 3 of 87 5.0 Public Health Data ...................................................................................................... 29 6.0 Exposure Characterization/ Assessment...................................................................... 29 6.1 Dietary Exposure/ Risk Pathway ......................................................................... 29 6.1.1 Residue Profile ....................................................................................... 29 6.1.2 Acute and Chronic Dietary Exposure and Risk........................................ 31 6.2 Water Exposure/ Risk Pathway ........................................................................... 34 6.3 Residential ( Non Occupational) Exposure/ Risk Pathway.................................... 34 7.0 Aggregate Risk Assessments and Risk Characterization ........................................... 34 7.1 Acute Aggregate Risk ........................................................................................ 34 7.2 Short, Intermediate, and Long Term Aggregate Risk....................................... 34 7.3 Cancer Risk........................................................................................................ 34 8.0 Cumulative Risk Characterization/ Assessment.......................................................... 34 9.0 Occupational Exposure/ Risk Pathway........................................................................ 35 10.0 Data Needs and Label Requirements.......................................................................... 35 10.1 Toxicology......................................................................................................... 35 10.2 Residue Chemistry ............................................................................................. 35 10.3 Occupational and Residential Exposure .............................................................. 36 11.0 Summary of Proposed and Recommended Tolerances .............................................. 36 References: .............................................................................................................................. 37 Appendices .............................................................................................................................. 38 Page 4 of 87 1.0 Executive Summary Benthiavalicarb isopropyl ( hereafter in this document referred to solely as benthiavalicarb) is a new valinamide carbamate type of fungicide being proposed for use on grapes and tomatoes ( both field and greenhouse) in the EU for the control of downy mildew. Its mode of action is through the inhibition of phospholipid biosynthesis. The R L stereoisomer of benthiavalicarb ( KIF 230RL is the main and pesticidally active component; however, the S L stereoisomer ( KIF 230S L), which is not pesticidally active, is also present as a minor impurity. The benthiavalicarb products being proposed for use in the EU on grapes and tomatoes include a 15% water dispersible granule ( WDG), for use only on tomatoes, and a multiple active ingredient WDG formulation containing both benthiavalicarb ( 1.75%) and mancozeb ( 70%) for use on both grapes and tomatoes. These formulations are labeled for multiple foliar applications. There are currently no food/ feed uses or tolerances for benthiavalicarb in the U. S.

The toxicology database for benthiavalicarb is complete and adequate for the purposes of risk assessment. None of the toxicity study endpoints were the result of a single oral exposure to benthiavalicarb. As a result, no acute dietary endpoint was identified and an acute dietary exposure assessment is not needed. The main target organ of toxicity is the liver in all four species tested ( rat, mouse, rabbit, and dog). Chronic exposure to benthiavalicarb also resulted in kidney toxicity in rats and mice. Prolonged exposure to benthiavalicarb resulted in the formation of several tumor types: liver tumors in male and female mice, thyroid follicular cell adenomas in male mice, and uterine tumors in rats. This compound is classified as " likely to be carcinogenic to humans." HED's Cancer Assessment Review Committee ( CARC) concluded that the mechanistic studies submitted for this chemical were insufficient to support definitive modes of action for the individual tumor types. Therefore, the Committee recommended using a low dose extrapolation approach ( Q1*) for estimating the human cancer risk based on the most potent tumor type ( liver tumors in mice).

Benthiavalicarb is not mutagenic, nor does this chemical produce developmental, reproductive, or neurotoxic effects.

Benthiavalicarb was absorbed and excreted rapidly in rats, with nearly full recovery of radioactive label within 168 hours. Analysis of the metabolites in bile, urine, and feces of cannulated rats identified a total of 13 compounds, including the parent, which accounted for 43 60% of the radiolabeled dose. Based on the components identified in excreta, the metabolism of benthiavalicarb in rats primarily involves either ( 1) hydroxylation of the phenyl ring followed by glucuronic acid conjugation or glutathione conjugation, or ( 2) hydroxylation of the phenyl ring followed by glutathione conjugation with subsequent degradation of the glutathione moiety, resulting in a variety of metabolites present in small quantities. Cleavage and hydroxylation of the valyl side chain also occurs to a limited extent. Parent ( excreted in feces only) and metabolites M15 ( urine and feces) and M18 ( urine only) were also the principal components detected in plasma, liver, and kidneys.

The risk assessment team concluded that none of the observed effects were the result of a single exposure to benthiavalicarb. As a result, an appropriate endpoint for acute dietary risk assessment is not available and an acute dietary exposure assessment is not needed. For chronic Page 5 of 87 dietary risk assessment, the endpoint selected is based on liver effects seen in the chronic rat study. The liver effects are the primary effects of concern. The NOAEL of 9.9 mg/ kg/ day is protective of tumorigenic effects observed at doses at, or above, 300 mg/ kg/ day ( uterine tumors in rats at, or above, 318 mg/ kg/ day and liver and thyroid tumors in mice at, or above, 358 mg/ kg/ day). The standard 100x uncertainty factor for inter and intra species variabilities is being used, and the special FQPA safety factor was reduced to 1x because of the absence of neurotoxic and developmental effects. Therefore, the chronic RfD and cPAD are 0.099 mg/ kg/ day. The target MOE is 100. For the purpose of cancer risk assessment the Q1* is 0.063 ( mg/ kg/ day) 1. This value was based on increases in male mouse liver combined adenomas, carcinomas, and/ or blastomas.

For purposes of this risk assessment, the nature of the residue in plants has been adequately delineated based on grape, tomato, and potato metabolism studies. In grapes and tomatoes, the metabolism of benthiavalicarb was minimal with parent being the only major residue in fruit and foliage. Parent was also the principal 14C residue in potato foliage, but other minor metabolites were identified. In potatoes, the metabolism of benthiavalicarb occurs primarily via direct hydroxylation of the phenyl ring or defluorination and hydroxylation of the phenyl ring. Secondary metabolism occurs via conjugation of sugars with hydroxy groups. Although two major (> 10%) residues were observed in potato tubers, the absolute levels of these metabolites ( suggested to be sugar conjugates) were < 0.006 ppm. Chiral analysis of potato foliage extracts also indicates that there was no isomeric conversion of the active R L isomer to other possible isomers ( S L, R D, or S D). Based on the results of the metabolism studies, the residues of concern in grapes and tomatoes for both tolerance expression and risk assessment are the combined residues of the R L and S L isomers of benthiavalicarb. The nature of the residue in livestock was not determined, and is not required at this time, as there are no significant livestock feed items associated with grape and tomato commodities.

An adequate GC/ NPD Method ( RCC No. 665943) is available for collecting data on residues of both the R L and S L isomers of benthiavalicarb in/ on grape and tomato commodities. For this method, residues are extracted with acetone, partitioned into hexane, purified using an amino SPE cartridge, and determined by GC/ NPD. The lower limit of method validation ( LLMV) is 0.01 ppm for each analyte in grape and tomato matrices. The limit of detection ( LOD) in grapes is 0.001 ppm. The LOD in grape processed commodities, tomatoes, and tomato processed commodities is 0.002 ppm. The method was adequately validated by the petitioner, and is being proposed as the tolerance enforcement method. The method was forwarded to the Analytical Chemistry Branch ( ACB) of the Biological and Economic Assessment Division ( BEAD) for a petition method validation ( PMV) trial. ACB concluded that the method does not need to be laboratory validated by EPA, and appears to be suitable for tolerance enforcement. As the method is not highly specific, the petitioner should either develop a confirmatory method for the analytes of interest or conduct an interference study for any future tolerances.

Adequate storage stability data are available indicating that benthiavalicarb is stable under frozen (  18 ° C) conditions for at least 13 months in grapes and 8 months in tomatoes. These data support the maximum storage intervals ( 7 8 months) from the grape and tomato field trials.

As there are no significant animal feed items associated with the proposed use on grapes and tomatoes, livestock feeding studies are not required for this petition. Tolerances do not need to Page 6 of 87 be established on animal commodities.

The available field trial data are adequate and will support the proposed use patterns on grapes and tomatoes. For grapes, 34 field tests were conducted throughout the EU in which grapes were treated at 1x the maximum proposed rate and harvested at the minimum PHI of 28 days. For tomatoes, 12 tests were conducted at the 1x application rate throughout the EU on both greenhouse and outdoor field grown tomatoes. The recommended tolerances for grapes and tomatoes were generated using HED's statistical tolerance generator for NAFTA harmonized tolerances. The recommended tolerance for grapes is 0.25 ppm, even though the highest field trial value is 0.27 ppm. The recommended tolerance for tomatoes is 0.45 ppm.

The available processing data are adequate. A separate tolerance is required for raisins. HED recommends in favor of the proposed tolerance of 1.0 ppm for grape, raisin. As residues do not concentrate in wine, the 0.25 ppm grape tolerance is adequate to cover residues in wine. A separate tolerance for wine is not needed. The tomato processing studies demonstrate that separate tolerances are not required for processed tomato fractions. Storage stability data were not submitted for processed grape and tomato commodities. These data are not required for the current tolerance requests because acceptable storage stability data were submitted for grapes and tomatoes. Residues were stable in grapes for 13 months and in tomatoes for 8 months. In the tomato storage stability studies, tomatoes were homogenized prior to storage. Homogenized tomatoes are comparable in both composition and texture to processed tomato commodities. The matrix is an acidic one which would very likely maximize residue decay. As a result, the tomato data are considered adequate to demonstrate storage stability in processed grape commodities as well.

As the proposed uses are for grapes and tomatoes grown in the EU, confined and field rotational crop studies are not required for the current petition.

A chronic dietary risk assessment was conducted using the Dietary Exposure Evaluation Model ( DEEM FCID, Version 2.03), which uses food consumption data from the USDA's Continuing Surveys of Food Intakes by Individuals ( CSFII) from 1994 1996 and 1998. The result of the chronic dietary exposure assessment for the general U. S. population is being used along with the cancer Q1* value to estimate the cancer risk to the U. S. population. The chronic dietary exposure analysis is based on average field trial values and estimates of percent crop imported. Empirical processing factors were used for most processed commodities. When empirical factors were not available, DEEM ( Version 7.81) default processing factors were used. The general U. S. population and all population subgroups have risk estimates that are below HED's level of concern ( i. e., 100% of the chronic population adjusted dose ( cPAD)). The risk estimates for all population subgroups are < 1% cPAD.

In an unrefined cancer dietary exposure analysis, the cancer risk to the general U. S. population exceeds HED's level of concern ( i. e., 1.0 x 10 6 ( or one in one million)). As a result, refinements were made to the analysis. The refinements include the use of average field trial values and estimates of percent crop imported. With the use of these refinements, the lifetime cancer risk to the U. S. population is 1.6 x 10 6. Although refinements were made to the analysis, it is considered to be conservative. First, average field trial values from field trials almost always exceed the expected residue levels found on crops at the time of consumption. When field trials are Page 7 of 87 performed, the maximum allowable application rate is used and crops are harvested at the minimum PHI. Samples are stored frozen until analysis to ensure minimal degradation of residues. In actual practice, however, growers will not usually use the maximum application rates for economic reasons. In addition, most crops are not harvested and immediately stored frozen. From previous dietary exposure analyses, HED has noted that monitoring data are often one to two orders of magnitude lower than field trial data. The second reason this analysis is considered to be conservative is that the registrant is planning on marketing benthiavalicarb in Europe only. In the dietary analysis, percent crop imported estimates were used for all imports, not just those from Europe. The third reason this analysis is considered to be conservative is that HED assumed that 100% of the imported crop will be treated. The actual percent crop treated will very likely be considerably lower than that. For these reasons, HED is confident that this analysis does not underestimate risk to the general U. S. population. HED is also confident that average field trial values and percent crop imported values used in the assessment will not be exceeded in the future. For the reasons stated above, the field trial values considerably overestimate residues found on food at the time of consumption. It is unlikely that average residue levels in imported grape and tomato commodities at the time of consumption would ever exceed the average field trial values. As far as percent crop imported values being exceeded in the future, this situation is unlikely as well. As stated above, the conservative assumptions were made that 100% of the grape and tomato commodities imported from all over the world would be treated. It's very unlikely that the quantities of imports actually treated will ever increase to those levels. Data for grape commodities were provided by BEAD. The economist who provided the data stated that it was unlikely the estimates of import shares would be exceeded in the near future. The data for grape commodities provided to HED were five year averages, and were compared to the annual data points of each data source, when available. Import share data do vary from year to year, but crop production patterns and consumption and preferences are more stable and change more slowly. For tomatoes, the worldwide imports changed as follows from 1998 to 2004: 37%, 31%, 30%, 33%, 32%, 37%, and 36%. Clearly, no increase occurred over that time period. For processed tomato commodities, the worldwide imports changed as follows from 2000 to 2004: 2.9%, 5.6%, 7.1%, 5.4%, and 5.8%. Between 2001 and 2004 there was no significant increase in processed tomato commodity imports.

The tolerances being granted in conjunction with these requests are for imported commodities only. As a result, residues in water are not being considered in this assessment. In addition, there are no residential uses for benthiavalicarb ( including home uses, recreational uses, spray drift, etc.); therefore, short term and intermediate term aggregate risk assessments are not needed. Because of the proposed use pattern, the chronic aggregate risk assessment is equivalent to the chronic dietary exposure analysis. The general U. S. population and all population subgroups had risk estimates that were well below HED's level of concern.

The aggregate cancer risk assessment is equivalent to the cancer dietary exposure analysis. The lifetime cancer risk to the U. S. population is 1.6 x 10 6. As stated above, the analysis is considered to be a conservative one. HED believes that the cancer dietary exposure analysis almost certainly overestimates exposure. HED's rationale for this belief is given in the discussion of cancer dietary exposure. The cancer risk to the general U. S. population does not exceed HED's level of concern. Page 8 of 87 No major deficiencies were noted in the subject petition that would preclude establishing tolerances for benthiavalicarb on grape and tomato commodities. However, before any further tolerances are granted, the petitioner should either develop a confirmatory analytical method for the analytes of interest or conduct an interference study. The recommended tolerances are as follows: 0.25 ppm for grape, 0.45 ppm for tomato, and 1.0 ppm for grape, raisin. The recommended tolerances were generated using HED's statistical tolerance generator for NAFTAharmonized tolerances. The residues to be included in the tolerance expression are the combined R L and S L isomers of benthiavalicarb. The petitioner has recommended that only the R L isomer be included in the tolerance expression because the S L isomer is a minor impurity in the commercial formulations and the potato metabolism study indicates that the active R L isomer is not converted to the S L isomer. In addition, the residue definition proposed for the EU MRLs includes only the R L isomer. The registrant has proposed EU MRLs of 0.2 ppm for grapes and 0.3 ppm for tomatoes. It's possible that if EU MRLs are established, the residue definition might be benthiavalicarb isopropyl only, and no distinction will be made as to the isomer. HED concludes that the S L isomer should be included in the tolerance expression, however. The results of the field trials indicate that the residue levels of the S L isomer are often non negligible compared to the levels of the R L isomer. In the grape field trials several samples had S L isomer levels that were 10 20% of the R L isomer levels. A small number of samples had S L: R L ratios that exceeded 20%. In the tomato field trials, the S L residue levels were generally nondetectable In a few cases, however, the S L isomer was present at 6 7% of the level of the R L isomer. The technical material contains 93% R L isomer and 3.4% S L isomer. The analytical method determines both isomers separately.

HED concludes that there are no residue chemistry or risk issues that would preclude establishing tolerances for the combined residues of isopropyl[( S) 1[[[( 1R) 1( 6 fluoro 2 benzothiazolyl) ethyl] amino] carbonyl] 2 methylpropyl] carbamate and isopropyl[( S) 1 [[[( 1S) 1( 6 fluoro 2 benzothiazolyl) ethyl] amino] carbonyl] 2 methylpropyl] carbamate at 0.25 ppm in/ on grape, 0.45 ppm in/ on tomato, and 1.0 ppm in/ on grape, raisin. As residues do not concentrate in wine, a separate tolerance for wine is not needed.

EPA has not made a common mechanism of toxicity finding as to benthiavalicarb and any other substances, and benthiavalicarb does not appear to produce a toxic metabolite produced by other substances. As a result, for the purposes of this tolerance action, EPA has not assumed that benthiavalicarb has a common mechanism of toxicity with other substances. Although benthiavalicarb is a carbamate compound, it is not a member of the class of insecticides known as the N methyl carbamates for which the Agency is presently conducting a cumulative risk assessment. The substituents on the benthiavalicarb nitrogen atom are much larger than the methyl group in the insecticides. While the N methyl carbamates are neurotoxicants based on their ability to inhibit the enzyme cholinesterase, there is no evidence of neurotoxicity or neuropathology in the hazard database for benthiavalicarb. Benthiavalicarb is also not a member of the thiocarbamate class of herbicides or the dithiocarbamate class of fungicides.

As benthiavalicarb will be used outside the U. S. to treat grapes and tomatoes, there will be no occupational risk to U. S. workers. As a result, an occupational assessment is not relevant to this tolerance petition. Page 9 of 87 2.0 Ingredient Profile Benthiavalicarb is a new valinamide carbamate type of fungicide being proposed for use on grapes and tomatoes ( both field and greenhouse) in the EU for the control of downy mildew. Its mode of action is through the inhibition of phospholipid biosynthesis. The R L stereoisomer of benthiavalicarb ( KIF 230R L) is the main and pesticidally active component; however, the S L stereoisomer ( KIF 230S L), which is not pesticidally active, is also present as a minor impurity.

The benthiavalicarb products being proposed for use in the EU on grapes and tomatoes include a 15% water dispersible granule ( WDG), for use only on tomatoes, and a multiple active ingredient WDG formulation containing both benthiavalicarb ( 1.75%) and mancozeb ( 70%) for use on both grapes and tomatoes. These formulations are labeled for multiple foliar applications.

2.1 Summary of Registered/ Proposed Uses Table 2.1. Summary of Directions for Use of Benthiavalicarb Applic. Timing, Type, and Equip. Formulation Applic. Rate ( lb ai/ A) Max. No. Applic. per Season Max. Seasonal Applic. Rate ( lb ai/ A) PHI ( days) Use Directions and Limitations Grapes Broadcast foliar application Ground or aerial equipment 1.75% WDG 1 0.031 6 0.187 28 Minimum application volumes and RTIs were not specified.

Tomatoes Broadcast foliar application Ground or aerial equipment 1.75% WDG 1 15% WDG 0.067 ( 0.031) 2 6 0.40 ( 0.19) 2 greenhouse  3 field  7 Minimum application volumes and RTIs were not specified.

1 The 1.75% WDG is a MAI that also contains 70% mancozeb. 2 When used in conjunction with mancozeb, the maximum single and seasonal application rate is lower.

2.2 Structure and Nomenclature Table 2.2. Nomenclature of Benthiavalicarb Compound S N N H H CH 3 NH O C H 3 CH 3 O H CH 3 C H 3 F O Page 10 of 87 Table 2.2. Nomenclature of Benthiavalicarb Common name Benthiavalicarb ( R L isomer)

Company experimental names KIF 230R L IUPAC name Isopropyl[( S) 1{[( 1R) 1( 6 fluoro 1,3 benzothiazol 2 yl) ethyl] carbamoyl} 2 methylpropyl] carbamate CAS name Isopropyl[( S) 1[[[( 1R) 1( 6 fluoro 2 benzothiazolyl) ethyl] amino] carbonyl] 2 methylpropyl] carbamate CAS # 177406 68 7 End use products/ EP 15% WDG, 1.75 WDG ( MAI with 70% Mancozeb)

Compound S N N H CH 3 H NH O C H 3 CH 3 O H CH 3 C H 3 F O Common name Benthiavalicarb ( S L isomer)

Company experimental names KIF 230S L IUPAC name Isopropyl[( S) 1{[( 1S) 1( 6 fluoro 1,3 benzothiazol 2 yl) ethyl] carbamoyl} 2 methylpropyl] carbamate CAS name Isopropyl[( S) 1[[[( 1S) 1( 6 fluoro 2 benzothiazolyl) ethyl] amino] carbonyl] 2 methylpropyl] carbamate CAS # Not available 2.3 Physical and Chemical Properties Table 2.3. Physicochemical Properties of Benthiavalicarb Parameter Value Reference Melting point 167 ± 0.5 ° C pH 4.35 at 25 ° C ( 1% w/ v aqueous suspension)

Density 1.25 g/ cm3 at 20.5 ± 0.5 ° C Water solubility ( 20 ° C) 10.96 mg/ L at pH 5 13.14 mg/ L at unadjusted pH 12.76 mg/ L at pH 9 Solvent solubility ( g/ L at 20 ° C) Not reported Vapor pressure at 25 ° C < 3.0 x 10 4 Pa Dissociation constant ( pKa) No dissociation from pH 1.12 12.81 at 20 ° C Octanol/ water partition coefficient Log( KOW) 2.52 2.59 at pH 5 9 UV/ visible absorption spectrum (  max, nm) Not reported Section A of PP# 3E6545 Page 11 of 87 3.0 Metabolism Assessment 3.1 Comparative Metabolic Profile The metabolism of benthiavalicarb in rats primarily involves either ( 1) hydroxylation of the phenyl ring followed by glucuronic acid conjugation or glutathione conjugation, or ( 2) hydroxylation of the phenyl ring followed by glutathione conjugation with subsequent degradation of the glutathione moiety resulting in a variety of metabolites present in small proportions. Cleavage and hydroxylation of the valyl side chain also occurs to a limited extent. In grapes and tomatoes, the metabolism of benthiavalicarb was minimal with parent being the only major residue in fruit and foliage ( 55 95% TRR). Parent was also the principal 14C residue in potato foliage ( 88 90% TRR), but other minor metabolites were identified. In potatoes, the metabolism of benthiavalicarb occurs primarily via direct hydroxylation of the phenyl ring or defluorination and hydroxylation of the phenyl ring. Secondary metabolism occurs via conjugation of sugars. Chiral analysis of potato foliage extracts also indicates that there was no isomeric conversion of the active R L isomer to other possible isomers ( S L, R D, or S D). Based on the results of the metabolism studies, the residues of concern in grapes and tomatoes for both tolerance expression and risk assessment are the combined residues of the R L and S L isomers of benthiavalicarb. The nature of the residue in livestock was not determined, and is not required, as there are no significant livestock feed items associated with grapes and tomatoes.

The metabolism of benthiavalicarb is more extensive in rats than it is in grapes, tomatoes, and potatoes. In all plant species tested, metabolism of parent benthiavalicarb occurred to a very limited extent. In both plants and animals, the primary metabolic pathways are hydroxylation of the phenyl ring with subsequent conjugation with sugars or glutathione. The only metabolic pathway occurring in plants that did not occur in rats was the defluorination that occurred in potatoes.

3.2 Nature of the Residue in Foods 3.2.1. Description of Primary Crop Metabolism In grapes and tomatoes, the metabolism of benthiavalicarb was minimal with parent being the only major residue in fruit and foliage ( 55 95% TRR). Parent was also the principal 14C residue in potato foliage ( 88 90% TRR), but other minor metabolites were identified. In potatoes, the metabolism of benthiavalicarb occurs primarily via direct hydroxylation of the phenyl ring or defluorination and hydroxylation of the phenyl ring. Secondary metabolism occurs via conjugation of sugars. Chiral analysis of potato foliage extracts also indicates that there was no isomeric conversion of the active R L isomer to other possible isomers ( S L, R D, or S D). Based on the results of the metabolism studies, the residues of concern in grapes and tomatoes for both tolerance expression and risk assessment are the combined residues of the R L and S L isomers of benthiavalicarb.

In grapes, the total radioactive residues ( TRR) were 0.241 ppm in/ on grapes and 21.66 ppm in/ on grapevine foliage from the [ Val 14C] test and 0.327 ppm in grapes and 14.01 ppm in foliage from the [ Bz 14C] test. HPLC analyses identified parent ( 94.0 96.5% TRR) as the main component in both fruit and foliage. No other metabolites were identified, and isolated unknowns accounted for Page 12 of 87   2.0% of the TRR in each matrix. Enzymatic hydrolysis of foliage extracts with   glucosidase did not show evidence of glucoside conjugation. In tomatoes, HPLC analyses identified parent as comprising 54.8 95.1% of the TRR in tomatoes and tomato foliage. Parent was the only compound detected in either fruit or foliage. In the 56 DAT fruit, unknowns accounted for a total of 40.9% of the TRR, but each unknown accounted for   9.4% of the TRR (< 0.001 ppm).

In potatoes, after a single foliar application to immature plants, parent ( 65 75% TRR) was the only component detected in foliage 14 DAT. Following a single postemergence soil application, 14C residues in tubers were too low for analysis, and the major component detected in foliage ( 90 DAT) was the sugar conjugate Metabolite 1 ( 28 30% TRR). Parent also accounted for 10 11% of the TRR in foliage and Metabolites 2, 3, and 6 each accounted for 3 17% of the TRR ( 0.001 0.011 ppm). Following the last of six broadcast foliar applications, the primary component identified in foliage at 14 DAT was parent ( 88 90% TRR), along with minor amounts of Metabolites 1, 2, and 6 ( 0.8 3.2% TRR). Treatment with   glucosidase and LC/ MS analysis identified Metabolites 1, 2, and 6, and sugar conjugates of ring hydroxylated metabolites. In tubers, the primary metabolites detected were Unknown Metabolites 4 ( 40% TRR) and 5 ( 23% TRR), along with trace amounts of parent ( 4.7% TRR). Treatment with   glucosidase suggests that Metabolites 4 and 5 are also sugar conjugates; however, concentrations of these metabolites were too low (< 0.006 ppm) for additional analysis.

3.2.2 Description of Livestock Metabolism The raw agricultural commodities ( RAC) and processed commodities associated with the proposed uses on imported grapes and tomatoes will not be used as significant animal feed items. Therefore, a determination of the nature of the residue in livestock is not required for this petition.

3.2.3 Description of Rotational Crop Metabolism, including identification of major metabolites and specific routes of biotransformation The proposed uses are for crops grown in Europe for importation to the U. S.; therefore, rotational crop studies are not required for this petition.

3.3 Environmental Degradation The tolerances being granted in conjunction with these requests are for imported commodities only. As a result, residues in water are not being considered in this risk assessment.

3.4 Toxicity Profile of Major Metabolites and Degradates In grapes, tomatoes, and potato foliage, parent benthiavalicarb was the principal 14C residue. In potato foliage, minor metabolites were identified. In rats, there were no major metabolites. In plants and rats, the minor metabolic products that formed include hydroxylated compounds in which the OH group appears on the phenyl ring and to a small extent on the valyl side chain. Other metabolites include sugar and glutathione conjugates ( of the phenyl ring). It is unlikely that these hydroxy metabolites, sugar conjugates, and glutathione conjugates will be more toxic than the parent compound itself. Page 13 of 87 3.5 Summary of Residues for Tolerance Expression and Risk Assessment 3.5.1 Tabular Summary Table 3.5. Summary of Metabolites and Degradates to be Included in the Risk Assessment and Tolerance Expression Matrix Residues included in Risk Assessment Residues included in Tolerance Expression Primary Crop Parent Benthiavalicarb Isopropyl, both R L and SL isomers Parent Benthiavalicarb Isopropyl, both R L and SL isomers Plants Rotational Crop N/ A N/ A Ruminant N/ A N/ A Livestock Poultry N/ A N/ A Drinking Water N/ A N/ A 3.5.2. Rationale for Inclusion of Metabolites and Degradates Based on the results of the metabolism studies, the residues of concern in grapes and tomatoes for both tolerance expression and risk assessment are the combined residues of the R L and S L isomers of benthiavalicarb. In grapes and tomatoes, the metabolism of benthiavalicarb was minimal with parent being the only major residue in fruit and foliage ( 55 95% TRR). Parent was also the principal 14C residue in potato foliage ( 88 90% TRR), but other minor metabolites were identified. Chiral analysis of potato foliage extracts indicates that there was no isomeric conversion of the active R L isomer to other possible isomers ( S L, R D, or S D). Although two metabolites in potato tubers comprised 23% and 40% of the TRR, these residues were present at very low levels (< 0.006 ppm), and appear to be sugar conjugates. In plants and rats, the metabolic products include hydroxylated compounds in which the OH group appears on the phenyl ring and to a small extent on the valyl side chain. Other minor metabolites include sugar and glutathione conjugates ( of the phenyl ring). It is unlikely that these hydroxy metabolites, sugar conjugates, and glutathione conjugates will be more toxic than the parent compound itself. As a result, taking into account the low % TRR or low ppm levels of the metabolites, a risk assessment of the parent compound will be protective of the metabolites as well.

Both the R L and S L isomers should be included in the tolerance expression as well. The results of the field trials indicate that the residue levels of the S L isomer are often non negligible compared to the levels of the R L isomer. In the grape field trials several samples had S L isomer levels that were 10 20% of the R L isomer levels. A small number of samples had S L: R L ratios that exceeded 20%. In the tomato field trials, the S L residue levels were generally non detectable. In a few cases, however, the S L isomer was present at 6 7% of the Page 14 of 87 level of the R L isomer. The technical material contains 93% R L isomer and 3.4% S L isomer. The analytical method determines both isomers separately.

4.0 Hazard Characterization/ Assessment 4.1 Hazard Characterization The toxicology database for benthiavalicarb is complete, including the studies needed for assessment of the FQPA Safety Factor. The liver is the target organ of toxicity for benthiavalicarb. Increased absolute and relative liver weights were seen in four species ( rat, mouse, rabbit, dog), accompanied by hepatocyte hypertrophy and lesions. Chronic exposure resulted in kidney toxicity in rats and mice. During the carcinogenicity studies in mice, prolonged exposure to benthiavalicarb was linked to increased incidences of hepatocellular adenomas, carcinomas, and blastomas. Benthiavalicarb was also linked to thyroid tumors in male mice and uterine tumors in female rats. Benthiavalicarb is not mutagenic, nor does it produce adverse developmental, reproductive, or neurotoxic effects.

In acute oral toxicity studies, no mortalities were observed up to the limit dose of 2000 mg/ kg. This chemical is classified as Toxicity Category III. No other acute studies were available for this chemical.

Benthiavalicarb was administered to mice ( MRID 45835007), rats ( MRID 45835004), and dogs ( MRID 45835006) for 90 days via their diet. All three species had increased absolute and relative liver weights and hepatocyte hypertrophy at their respective LOAELs. The adverse effects observed included decreases in hematocrit, hemoglobin, serum albumin, and albumin/ globulin ratios in females and hemosiderin pigment deposits in the spleen of males at 200 mg/ kg/ day. At the LOAEL of 352 mg/ kg/ day in rats, increases in total cholesterol and GGT were evident in both sexes; free cholesterol and phospholipids were increased in the females; and total protein was increased in males. Body weight gains and food consumption were decreased in male and female mice dosed above 1293 mg/ kg/ day. In mice, liver lesions that occurred at or above a dose of 1293 mg/ kg/ day included black colored livers in females and necrosis, hypertrophy, and anisonucleosis in males.

In the chronic rat ( MRID 45835017) and mouse ( MRID 45835016) studies, the effects were found primarily in the liver. Both species had increased absolute and relative liver weights, hepatocyte hypertrophy, and gross liver lesions. The chronic study in dogs ( MRID 45835015) demonstrated minimal increases in absolute and relative liver weights. In mice, liver lesions included brown patches and nodules in both sexes, enlarged livers in females, intermediate fatty change, and hepatic foci of cellular alteration in both sexes. In male mice, anisonucleosis, necrosis, single cell necrosis, multinucleated cells, accumulation of macrophages, cellular infiltration, lymphocyte infiltration, bile duct proliferation, and extramedullary hematopoiesis were also present at the LOAEL of 358 mg/ kg/ day. Similar liver lesions were present in rats at doses greater than 250 mg/ kg/ day. In both sexes of mice, thyroid cell hyperplasia was present at the LOAEL. In males, dilatated thyroid follicles were present at the LOAEL. In the chronic rat study, the kidney was also a target organ of toxicity. The following kidney lesions were observed at the LOAEL of 250 mg/ kg/ day: chronic nephropathy and tubular dilatation in the males, Page 15 of 87 glomerulosclerosis; calculus and hyaline droplets in both sexes; lymphocytic cellular infiltration, basophilic tubules, hyaline casts, and brown pigment deposits in females.

Several tumor types were seen in the carcinogenicity studies in both rats ( MRID 45835017) and mice ( MRID 45835016). In the rat carcinogenicity study, animals were exposed to 0, 50, 200, 5,000, and 10,000 ppm of benthiavalicarb via dietary admix. Male rats in the high dose group had a statistically higher incidence of hepatocellular adenomas than did the control group. The incidence rate was within the range of historical control data; therefore, the CARC concluded that this tumor type in rats may not be directly related to exposure to benthiavalicarb. However, a similar increase in hepatocellular tumors occurred in mice, and the CARC concluded that the liver tumor data in rats may provide some supporting evidence of carcinogenic potential for liver tumors. In the mouse carcinogenicity study, animals were administered 0, 20, 100, 2500, or 5,000 ppm of benthiavalicarb in their diets. Male mice in the 2,500 and 5,000 ppm dose groups had statistically higher incidences of hepatocellular adenomas, carcinomas, blastomas, and combined tumors than controls. Female mice in the top two dose groups had increased incidences of hepatocellular adenomas. The increases in hepatocellular tumors in mice were considered to be treatment related, and this effect was used to calculate the Q1* for risk assessment. All doses in the rat and mouse studies were considered adequate, but not excessive, for determination of carcinogenic potential, except for the 5,000 ppm dose in mice, which was considered to be excessive because it resulted in severe systemic toxicity.

Female rats in the top two dose groups had a significantly increased incidence of uterine adenocarcinomas compared to concurrent controls. The incidence rate also exceeded the rates in historical control data. Therefore, uterine tumors in rats were considered to be treatment related. Similarly the increased incidence of follicular cell adenomas in male mice in the 2,500 and 5,000 ppm dose groups were considered to be treatment related. Follicular cell hyperplasia was also present in these dose groups.

Benthiavalicarb is classified as " likely to be carcinogenic to humans."

Eleven mechanistic studies were submitted in order to determine a mode of action for benthiavalicarb. After thorough consideration by the CARC, it was concluded that the data were insufficient to support definitive modes of action for each tumor type. Therefore, the CARC recommended using a linear low dose extrapolation approach ( Q1*) for estimating human cancer risk based upon the most potent tumor type. After quantitative analysis of the tumor types, the Q1* was based on combined hepatocellular tumors in male mice. The Q1* is 6.2795x10 2.

More than 20 mutagenicity studies were submitted for benthiavalicarb, testing the mutagenic potential of various lots of the parent and a number of congeners. All in vitro and in vivo studies, except two, were negative for the induction of mutagenicity up to the limit dose or precipitation of the compound. The two exceptions of positive results were with two lots of the parent compound in the TA98 strain of S. typhimurium in the presence of S9 activation. However, later studies demonstrated that these positive results were not due to the parent, but to a mutagenic impurity ordinarily at low, ineffective levels in all but these two batches.

During a metabolism study in rats ( MRID 45835108), both the low and high dose radiolabeled forms were absorbed rapidly, with nearly full recovery of radioactive label ( 89 100%) within 168 Page 16 of 87 hours. Radioactivity was recovered primarily in the feces ( 62 83%) and urine ( 7 24%), with up to 85% recovery in 48 hours. Urinary excretion was slightly higher in females than males. Absorption was greater in the low dose group than in the high dose group, indicating incomplete absorption in the high dose group. Radiolabeled residues preferentially partitioned into plasma. In both dose groups ( at all time points), the concentration of radioactivity was higher in the kidneys and liver than it was in plasma.

Analysis of the metabolites in bile, urine, and feces of cannulated rats identified a total of 13 compounds, including the parent, which accounted for 43 60% of the radiolabeled dose. The metabolite profile was similar between the sexes. A total of 11 metabolites were identified in the urine, most present in concentrations < 2% of the administered dose. A total of 8 metabolites were found in the bile, with the principal metabolites being isomers of hydroxylated glutathione conjugates and a glucuronide conjugate. Parent compound was not present in the urine or bile.

Based upon the components identified in excreta, the metabolism of benthiavalicarb in rats primarily involves either hydroxylation of the phenyl ring followed by glucuronic acid conjugation or glutathione conjugation and hydroxylation of the phenyl ring with subsequent degradation of the glutathione moiety, resulting in a variety of metabolites present in small quantities. Cleavage and hydroxylation of the valyl side chain also occurs to a limited extent. Parent ( excreted in feces only) and metabolites, M15 ( urine and feces) and M18 ( urine only), were also the principal components detected in plasma, liver, and kidneys.

Table 4.1. Acute Toxicity Profile  KIF 230 Guideline No. Study Type MRID( s) Results Toxicity Category 870.1100 Acute oral rat KIF 230 Tech: 45834909 KIF 230 S L: 45834910 KIF 230 1 KR: 45835011 KIF 230 1 1( S): 45835002 KIF 230 1 13: 45835003 LD50 > 2000 mg/ kg III 870.1200 Acute dermal [ species] NA 870.1300 Acute inhalation [ species] NA 870.2400 Acute eye irritation [ species] NA 870.2500 Acute dermal irritation [ species] NA 870.2600 Skin sensitization [ species] NA NA = not available Page 17 of 87 Guideline No./ Study Type MRID No. ( year)/ Classification / Doses Results Subchronic Studies 870.3100 90 Day oral toxicity rodents ( Fischer rats) 45835004 ( 1998) Acceptable/ Guideline 0, 50, 200, 7000, or 20,000 ppm M: 0, 3.5, 14.1, 353, or 1444 mg/ kg/ day F: 0, 3.9, 15.3, 379, or 1552 mg/ kg/ day NOAEL = 14.1/ 15.3 mg/ kg/ day LOAEL = 353/ 379 mg/ kg/ day based on hepatocyte hypertrophy in both sexes and increases in: absolute and relative to body liver weights, total cholesterol, and GGT in both sexes; free cholesterol and phospholipids in the females; and total protein in the males.

870.3100 90 Day oral toxicity rodents ( B6C3F1 mice) 45835007 ( 1998) Acceptable/ guideline 0, 50, 200, 7000, or 20,000 ppm M: 0, 8.4, 33.0, 1293, or 4031 mg/ kg/ day F: 0, 11.3, 45.2, 1620, or 4946 mg/ kg/ day NOAEL = 33.0/ 45.2 mg/ kg/ day LOAEL = 1293/ 1620 mg/ kg/ day based on decreased body weight, body weight gain, and food efficiency in males, increased absolute and relative liver weight, enlarged liver, black colored liver ( females only), and histopathological liver effects ( necrosis, hypertrophy, and anisonucleosis [ males only]).

870.3150 90 Day oral toxicity in dogs ( beagle) 45835005 ( 1999) Acceptable/ Guideline 0, 40, 200, or 1000 mg/ kg/ day NOAEL = 40 mg/ kg/ day LOAEL = 200 mg/ kg/ day based on decreases in hematocrit, hemoglobin, serum albumin and albumin/ globulin ratio in females and increases in liver to body weight ratios, hepatocyte hypertrophy ( both sexes) and hemosiderin pigment deposits in the spleen of males.

Developmental and Reproduction Studies 870.3700a Prenatal developmental in rodents ( Sprague Dawley) 45835012 ( 2000) Acceptable/ Guideline 0, 10, 100, or 1000 mg/ kg/ day Maternal NOAEL = 100 mg/ kg/ day LOAEL = 1000 mg/ kg/ day based on increased absolute and relative liver weights and on increased incidence of enlarged liver. Developmental NOAEL = 1000 mg/ kg/ day LOAEL = not observed.

870.3700b Prenatal developmental in rabbits ( New Zealand White) 45835010 ( 2000) Acceptable/ Guideline 0, 10, 20, or 40 mg/ kg/ day Maternal NOAEL = 20 mg/ kg/ day LOAEL = 40 mg/ kg/ day based on increased absolute and relative liver weights. Developmental NOAEL = 40 mg/ kg/ day LOAEL = not observed.

870.3800 Reproduction and fertility effects ( Sprague Dawley) 45835014 ( 1999) Acceptable/ Guideline 0, 100, 1000, or 10,000 ppm M: 0, 8.4, 84.1, or 880 mg/ kg/ day F: 0, 8.8, 91.0, or 943 mg/ kg/ day Parental/ Systemic NOAEL = 880/ 943 mg/ kg/ day LOAEL = not observed. Reproductive NOAEL = 880/ 943 mg/ kg/ day LOAEL = not observed. Offspring NOAEL = 880/ 943 mg/ kg/ day LOAEL = not observed.

Chronic and Carcinogenicity Studies Page 18 of 87 Guideline No./ Study Type MRID No. ( year)/ Classification / Doses Results 870.4100a Chronic toxicity rats ( Fischer) 45835017 ( 2001) Acceptable/ Guideline 0, 50, 200, 5000, or 10,000 ppm M: 0, 2.5, 9.9, 249.6, or 518.3 mg/ kg/ day F: 0, 3.2, 12.5, 318.2, or 649.4 mg/ kg/ day NOAEL = 9.9/ 12.5 mg/ kg/ day LOAEL = 249.6/ 318.2 mg/ kg/ day based on nephrotoxicity and hepatotoxicity in both sexes. In the kidney, absolute and relative weights were increased in both sexes. Kidney lesions included chronic nephropathy and tubular dilatation in males; glomerulosclerosis, calculus, and hyaline droplets in both sexes; lymphocytic cellular infiltration, basophilic tubules, hyaline casts, and brown pigment deposits in females. Liver findings included transient increased _ glutamyltranspep tidase in both sexes; increased levels of total and free cholesterol and phospholipids in females; increased absolute and relative liver weights in both sexes; increased incidences of brown and/ or white patches on the liver in males, and red patches on the liver in females; increased incidences of hepatocellular hypertrophy; and increased incidences of fatty liver in both sexes. At week 104, increased incidences of foci/ area of cellular alteration in the liver and spongiosis hepatis were observed in the males.

Increased incidences of hepatocellular tumors in males and uterine tumors in females were observed.

870.4100b Chronic toxicity dogs ( beagle) 45835015 ( 2001) Acceptable/ Guideline 0, 4, 40, or 400 mg/ kg/ day NOAEL = 40 mg/ kg/ day LOAEL = 400 mg/ kg/ day based on minimal increases in absolute and relative liver weights ( not significant) at the highest dose level.

870.4300 Carcinogenicity mice ( B6C3F1) 45835016 ( 2001) Acceptable/ Guideline 0, 20, 100, 2500, or 5000 ppm M: 2.7, 13.7, 358.4, or 731.3 mg/ kg/ day F: 0, 3.7, 18.6, 459.3, or 927.8 mg/ kg/ day NOAEL = 13.7/ 18.6 mg/ kg/ day LOAEL = 358.4/ 459.3 mg/ kg/ day based on clinical signs ( abdominal masses and distention, and pallor) in males, decreased body weight gain and food efficiency in males; gross liver lesions ( brown patches and nodules in both sexes, and enlarged livers in females); increased absolute and relative to body liver weight in both sexes; non neoplastic liver lesions ( hepatocyte hypertrophy, hepatic intermediate fatty change, hepatic foci of cellular alteration in both sexes, anisonucleosis, necrosis, single cell necrosis, multinucleated cells, accumulation of macrophages, cellular infiltration, lymphocyte infiltration, bile duct proliferation and extramedullary hematopoiesis in males); thyroid follicular cell hyperplasia in both sexes; and dilated thyroid follicles in males. no evidence of carcinogenicity Neurotoxicity Studies 870.6200a Acute neurotoxicity screening battery in rats ( Sprague Dawley) 45869735 ( 2002) Acceptable/ Guideline 0 or 2000 mg/ kg NOAEL = 2000 mg/ kg LOAEL = not observed Subchronic neurotoxicity study in rats ( CD rats) 45869736 ( 2002) Acceptable/ Guideline 0, 200, 2000, or 20,000 NOAEL = 174.1/ 185.7 mg/ kg/ day LOAEL = 1853.7/ 1845.8 mg/ kg/ day based on decreased body weight and body weight gains in the males. Page 19 of 87 Guideline No./ Study Type MRID No. ( year)/ Classification / Doses Results ppm M: 0, 17.7, 174.1, or 1853.7 mg/ kg/ day F: 0, 19.3, 185.7, or 1845.8 mg/ kg/ day Gene Mutation Studies Gene Mutation 870.5300 Mutation at the Thymidine Kinase Locus of Mouse Lymphoma Cells 45869728 ( 1999) Acceptable/ Guideline KIF 230 technical Negative for increases in mutant frequency up to the highest soluble concentration in the presence or absence of activation.

Gene Mutation Nonguideline Gene Mutation in Transgenic Mice 45869730 ( 2000) Acceptable/ Non guideline KIF 230 technical Negative for the induction of in vivo gene mutation in mice treated up to the limit dose.

Reverse Gene Mutation 870.5100 45869722 ( 1999) Acceptable/ Guideline KIF 230 technical Positive for gene mutation in the presence of rat liver activation when assayed up to 5000 _ g/ plate ( because of impurity).

Reverse Gene Mutation 870.5100 45835106 ( 2001) Acceptable/ Guideline KIF 230S L Negative for reverse gene mutation up to the limit of solubility.

Reverse Gene Mutation 870.5100 45869717 ( 2002) Acceptable/ Guideline KIF 230 Lot G51 47 190 Negative for inducing gene mutations in the presence and/ or absence of activation up to the limit dose of 5000 _ g/ plate.

Reverse Gene Mutation 870.5100 45869718 ( 2002) Acceptable/ Guideline KIF 230 Lot G51 48 190 Negative for inducing gene mutations in the presence and/ or absence of activation up to the limit dose of 5000 _ g/ plate.

Reverse Gene Mutation 870.5100 45869719 ( 2002) Acceptable/ Guideline KIF 230 Lot G51 49 190 Negative for inducing gene mutations in the presence and/ or absence of activation up to the limit dose of 5000 _ g/ plate.

Reverse Gene Mutation 870.5100 45869720 ( 2002) Acceptable/ Guideline KIF 230 Lot G51 50 190 Negative for inducing gene mutations in the presence and/ or absence of activation up to the limit dose of 5000 _ g/ plate.

Reverse Gene Mutation 870.5100 45869721 ( 2002) Acceptable/ Guideline KIF 230 Lot G51 51 190 Negative for inducing gene mutations in the presence and/ or absence of activation up to the limit dose of 5000 _ g/ plate.

Reverse Gene Mutation 870.5100 45869723 ( 2001) Acceptable/ Guideline KIF 230 Lot G51 35 184 Negative for inducing gene mutations in the presence and/ or absence of activation up to the limit dose of 5000 _ g/ plate.

Reverse Gene 45869724 ( 2001) Negative for inducing gene mutations in the presence and/ or Page 20 of 87 Guideline No./ Study Type MRID No. ( year)/ Classification / Doses Results Mutation 870.5100 Acceptable/ Guideline KIF 230 Lot G51 37 184 absence of activation up to the limit dose of 5000 _ g/ plate.

Reverse Gene Mutation 870.5100 45869725 ( 2001) Acceptable/ Guideline KIF 230 Lot G51 36 184 Negative for inducing gene mutations in the presence and/ or absence of activation up to the limit dose of 5000 _ g/ plate.

Reverse Gene Mutation 870.5100 45869726 ( 2001) Acceptable/ Guideline KIF 230 Lot G51 08 158 This lot had innate mutagenic activity in one bacterial strain in the presence of S9 activation when exposed to concentrations < 320 mg/ plate. In a repeat study, the positive response was demonstrated to be due to a mutagenic impurity ( 45835107)

Reverse Gene Mutation 870.5100 45869732 ( 2001) Acceptable/ Guideline KIF 230 I 1 ( R) Negative for inducing gene mutations in the presence and/ or absence of activation up to the limit dose of 5000 _ g/ plate.

Reverse Gene Mutation 870.5100 45839733 ( 2001) Acceptable/ Guideline KIF 230 I 1 ( S) Negative for inducing gene mutations in the presence and/ or absence of activation up to the limit dose of 5000 _ g/ plate.

Reverse Gene Mutation 870.5100 45869734 ( 2001) Acceptable/ Guideline KIF 230 I 13 Negative for inducing gene mutations in the presence and/ or absence of activation up to the limit dose of 5000 _ g/ plate.

Structural Chromosomal Aberration In vitro Chromosomal Aberration Test 870 5375 45869727 ( 1998) KIF 230 technical Unacceptable/ Not upgradeable There were no increased incidences in chromosomal aberrations at any dose compared to concurrent or historical controls. However this study is classified as unacceptable/ not upgradeable due to the presence of a " white powdery test substance" at all concentrations in this assay.

Other Genotoxic Effects In vivo/ In vitro Unscheduled DNA Synthesis 870.5550 45869731 ( 2001) Acceptable/ Guideline Negative for UDS induction in hepatocytes isolated from rats treated up to the limit dose, 2000 mg/ kg.

Measurement of Unscheduled DNA Synthesis in vitro 870.5550 45869729 ( 1999) Acceptable/ Guideline KIF 230 Negative for UDS induction in hepatocytes isolated from rats treated up to the limit dose, 2000 mg/ kg.

Induction of Micronuclei in the Bone Marrow of Treated mice 870.5385 45835105 ( 2000) Acceptable/ Guideline KIF 230 Negative for the induction of micronucleated polychromatic erythrocytes in bone marrow of mice treated up to the limit dose, 2000 mg/ kg.

Identification of a Mutagenic Substance Isolated from KIF 45835107 ( 2002) Summary The positive results with Lot Nos. G 51 15 162 and G51 08 158 in TA98 in the presence of S9 activation previously reported appears to be due to a mutagenic component identified Page 21 of 87 Guideline No./ Study Type MRID No. ( year)/ Classification / Doses Results 230 Nonguideline as 6,6' difluoro 2,2' bibenzothiazole.

Non Guideline Mechanistic Studies Hepatocarcinogenicity Study in Rats 45835021 ( 2000) Acceptable/ Non guideline Five groups of 12 male Fischer rats were used for this study. Groups 1, 3, and 5 were initiated by i. p. injection of diethylnitrosamine ( DEN) in 0.9% saline, while groups 2 and 4 were dosed with 0.9% saline only. Group 1 rats were fed a normal basal diet, while groups 2 and 3 received a diet with 10,000 ppm ( 586.5 634.1 mg/ kg/ day) KIF 230, and groups 4 and 5 received diets including 0.05% phenobarbital sodium. At week 3 a partial hepatectomy was performed to stimulate hepatic cell division No treatment related effects were observed on mortality or clinical signs. Minor decreases in body weights, cumulative body weight gains, and food efficiency were observed, but were not considered treatment related.

Rats in groups 2 and 3, which were dosed with KIF 230, had increased absolute and relative liver weights compared to controls. Enlarged liver, hepatocyte hypertrophy, and increased number of GST positive foci were seen in all treatment groups ( saline/ KIF 230, DEN/ KIF 230, saline/ PB, and DEN/ PB). Groups 4 and 5, which received PB, had increased incidence of fatty change in the liver. Increased mitosis was observed in 10/ 12 animals dosed with DEN/ KIF 230 and 12/ 12 animals dosed with DEN/ PB compared to 4/ 12 animals treated with DEN alone. Increased incidence of acidophilic cell focus was noted in 10/ 12 animals dosed with DEN/ KIF 230 and 9/ 12 animals dosed with DEN/ PB, compared to 4/ 12 animals treated with DEN alone.

Data indicate that administration of KIF 230 in the diet at 10,000 ppm to male rats caused a promoter effect in the liver similar to that caused by phenobarbital, a known tumor promoter in rat liver in a two stage experimental model of hepatocarcinogenesis.

Hepatocarcinogenicity Study Examining the Initiator Effect of KIF 230 in Male Rats 45869715 ( 2000) Acceptable/ Non guideline Male Fischer rats were placed into groups of 12 each. A partial hepatectomy was performed on all groups to stimulate cell division. Twelve hours later, the animals were initiated with a single injection of 20 mg/ kg DEN, a single gavage dose of 2000 mg/ kg KIF 230, or a single gavage dose of carboxymethyl cellulose sodium salt ( CMC Na). All rats were fed a basal diet containing phenobarbital sodium salt ( PB) from weeks 2 10 of the study. At week 3, all rats were given a single injection of 300 No treatment related signs were observed on mortality, clinical signs of toxicity, body weights, body weight gains, or gross pathology.

Absolute and relative liver weights were increased in the KIF 230 group compared to the DEN group. Fatty change was observed in all animals in all groups. Hepatocyte hypertrophy was seen in the DEN and KIF 230 groups. Mitosis was observed in 6/ 12 animals in the DEN group, compared to 0/ 12 and 1/ 12 animals in the KIF 230 and CMC Na groups, respectively. Clear cell foci ( 3/ 12) and eosinophilic cell foci ( 8/ 12) were observed in the DEN group compared to 0/ 12 animals in the KIF 230 or CMC Na groups. The number of GST P positive foci and the area of the GST P positive foci were greater in the DEN groups compared to the KIF 230 and CMC Na groups.

The data indicated that administration of KIF 230 as a single dose of 2000 mg/ kg to male rats did not demonstrate an initiator effect in the liver when compared to DEN, a known initiating agent in the two stage hepatocarcinogenic model. However, an additional group of saline/ PB treated rats should have been included in this study to allow for full comparison of treated Page 22 of 87 Guideline No./ Study Type MRID No. ( year)/ Classification / Doses Results mg/ kg D galactosamine. animals with untreated controls.

Two stage Transformation Assay 45835103 ( 2000) Acceptable/ Non guideline Initiation: BALB/ c 3T3 A31 1 1 cells were exposed to KIF 230 at concentrations of 0, 10.4, 17.3, 28.8, 48.0, or 80.0 g/ mL for three days. Promoter: cells were exposed to KIF 230 at concentrations of 0, 3.0, 6.0, 9.0, 12.0, or 15.0 g/ mL for three days. The test compound did not act as an initiating or promoting agent under conditions of this study.

Mechanism Study of Thyroid Tumors in Rats 45835018 ( 2002) Acceptable/ Non guideline 12 male Fisher rats/ sex/ dose were fed KIF 230 in their diets at concentrations of 0, 13.3, or 661.4 mg/ kg/ day for 14 days. The data indicated that the administration of KIF 230 in the diet at a dose of 661.4 mg/ kg/ day resulted in decreased serum T4 levels ( 11 18%) in male rats. Absolute and relative to body liver weights were increased ( 22 24%) compared to controls, and macroscopic examination revealed enlarged liver in all rats in this dose group, which is consistent with enzyme induction. TSH levels were non significantly increased compared to controls, and T3 levels remained similar to controls. The observed increase in UDP GT activity ( 16%) on T4 could cause a decrease in circulating T4 levels, resulting in increased serum TSH levels via a feedback mechanism. This mechanism could ultimately cause hyperthyroidism following a sufficient duration of exposure in rats.

Mechanism Study of Thyroid Tumors in Mice 45835019 ( 2002) Acceptable/ Non guideline 12 Scl: B6C3F1 male mice/ dose were exposed to KIF 230 in their diet at concentrations of 0, 17.0, or 855.0 mg/ kg/ day for 14 days. No effects were observed on mortality, clinical observations, body weights, cumulative body weight gains, or food consumption.

Increased absolute and relative liver weights were observed at 855 mg/ kg/ day, which is consistent with enzyme induction and the increased UDP GT activity on T4 seen in this study. While serum T4 levels were decreased 25 29%, T3 and TSH remained similar to controls throughout the study.

Typical UDP GT inducers are known to cause hyperthyroidism by inducing TSH through a decrease in serum thyroid hormone levels; and thyroid hyperplasia is a potential mechanism for the induction of thyroid tumors.

The lack of an appropriate antibody ( anti mouse TSH) could cause inaccurate measurements of TSH in the radioimmunoassay used to determine serum levels of TSH.

While the mechanism is plausible, no definitive conclusions can Page 23 of 87 Guideline No./ Study Type MRID No. ( year)/ Classification / Doses Results be reached concerning this mechanistic study.

Drug  Metabolizing Enzymes and Hepatocyte Proliferation in Mice 45835102 ( 2001) Acceptable/ Non guideline Eight B6C3F1 mice / sex/ dose were exposed to KIF 230 by gavage at concentrations of 0, 10, or 1000 mg/ kg/ day for seven days. Five mice/ sex/ dose were then checked for hepatocyte P 450 and specific P 450 isoenzyme levels. Three mice/ sex/ dose were evaluated for hepatocyte proliferation. No effects were observed on mortality, clinical observations, body weights, cumulative body weight gains, or on the BrdU labeling index.

At 1000 mg/ kg/ day, absolute and relative liver weights were increased 13 31%. Microscopic examination revealed hypertrophy of the hepatocytes in the liver of both males and females compared to controls. Total microsomal P 450 levels were increased ( 71 92%) in both sexes, CYP1A2 was increased 63 149%, CYP2B1 ( 2B2) increased 180 553%, and CYP3A2 increased 104 172%. CYP1A1, CYP2E1, and CYP4A1 levels were similar to control.

The increased liver weights and increased liver enzymes are consistent with enzyme induction and hepatocyte hypertrophy. The level of enzyme induction observed was low compared to the induction seen with classical induction agents like phenobarbital.

Drug  Metabolizing Enzymes and Hepatocyte Proliferation in Rats 45835101 ( 2001) Acceptable/ Non guideline Eight Fischer rats/ sex/ dose were exposed to KIF 230 by gavage at concentrations of 0, 10, or 1000 mg/ kg/ day for seven days. Five rats/ sex/ dose were then checked for hepatocyte P 450 and specific P 450 isoenzyme levels. Three mice/ sex/ dose were evaluated for hepatocyte proliferation. No effects of treatment were observed on mortality, clinical observations, body weights, cumulative gains, gross or microscopic pathological examinations, or on the hepatic BrdU labeling index.

At 1000 mg/ kg/ day, absolute and relative liver weights were increased 8 22%. In males, total hepatic microsomal P 450 levels were increased 18%, and several specific CYP enzymes were increased101 160%. In females, specific CYP enzymes were increased 76 130%. Levels of CYP2E1 and CYP4A1 were similar to controls in both sexes.

KIF 230 at 1000 mg/ kg/ day for seven days resulted in increased liver weights and enzyme induction. Unchanged levels of BrdU indicate that there was no treatment related effect on hepatocyte proliferation. The level of P 450 induction in males was greater than in females, but the magnitude of difference does not clearly indicate a sex difference.

Oxidative DNA in Mice 45835022 ( 2001) Acceptable/ Non guideline Five Scl: B6C3F1 mice/ sex/ dose were exposed to KIF 230 in their diets at concentrations of 0, 100, or 5000 ppm ( 0, 19.4/ 26.1, or 1031.2/ 1203.7 mg/ kg/ day male/ female) for up to 2 weeks. No effects of treatment were observed on mortality, clinical observations, body weights, cumulative body weight gains, or food consumption.

At 5000 ppm, absolute and relative body weights were increased ( 75 78%). No effects were observed on 8 OHdG levels measured in the liver tissue DNA.

Liver enlargement was consistent with enzyme induction. There was no indication of hepatic oxidative DNA damage, as measured by 8 OHdG production. Page 24 of 87 Guideline No./ Study Type MRID No. ( year)/ Classification / Doses Results Oxidative DNA Damage in Rat Liver 45869716 ( 2001) Acceptable/ Non guideline Five Fischer rats/ sex/ dose were exposed to KIF 230 in their diets at concentrations of 0, 200, or 10,000 ppm ( 0, 17.4/ 17.1, or 797.5/ 914.5 mg/ kg/ day male/ female) for up to 2 weeks. No treatment related effects were observed on mortality or clinical signs.

At 10,000 ppm, absolute and relative liver weights were increased ( 73 113%) in both sexes. At 200 ppm, relative liver weights were increased slightly in females ( 9%). 8 OHdG levels were decreased in the 200 and 10,000 ppm males ( 20% each)

Body weights and cumulative body weight gains were decreased ( 12 50%) in the high dose males. Decreased food consumption was also observed in these animals.

Liver enlargement was consistent with enzyme induction. There was no indication of hepatic oxidative DNA damage, as measured by 8 OHdG production.

Mechanism Study of Uterine Cancer in Rats 45835020 ( 2002) Acceptable/ Non guideline 12 Fischer female rats/ dose were exposed to KIF 230 in the diet at concentrations of 0, 200, or 10,000 ppm ( 0, 11.6, or 576.4 mg/ kg/ day) for up to 8 weeks. Microsomal aromatase activity was measured at week 8, while serum estradiol ( E2), progesterone ( P4), and luteinizing hormone ( LH) levels were measured at weeks 2, 4, 6, and 8. No treatment related effects were observed on mortality, clinical signs of toxicity, body weight, cumulative body weight gains, or food consumption.

At 576.4 mg/ kg/ day, microsomal aromatase activity was increased in the liver 36% and slightly increased in the uterus ( 15%), but was decreased in the ovary (  10%). No treatmentrelated effects were observed on serum sex hormone levels. Ovary and uterine weights were unaffected by treatment, and no effects were noted during necropsy. Absolute and relative liver weights were increased ( 28 31%), and macroscopic examination revealed dark liver in 10/ 11 rats and enlarged liver in 11/ 11 rats compared to 0/ 10 controls.

The data indicate that KIF 230 in the diets of female rats at 10,000 ppm does cause increased absolute and relative liver weights and increased aromatase activity in the liver, but it does not appreciably alter aromatase levels in organs other than the liver. Results were consistent with enzyme induction, which could result in upregulation of hepatic aromatase, but this increase was insufficient to increase circulating E2 levels in this study. The lack of increased aromatase activity in the ovary and uterus suggested little direct participation of E2 in the development of uterine cancer by KIF 230.

Uterotrophic Effects in Ovariectomized Rats 45869714 ( 2001) Unacceptable/ Nonguideline Six ovariectomized rats per dose were exposed to KIF 230 daily by gavage for 14 days at doses of 0, 10, 100, or 1000 mg/ kg/ day. A positive control group was dosed with 0.001 No treatment related effects were observed on mortality, clinical signs, body weight, overall body weight gains, absolute or relative uterine weights, microscopic uterine pathology, or uterine cellular proliferation as determined by BrdU incorporation.

The lack of a uterotropic response in the positive controls makes it impossible to determine if the lack of response in the test compound treated animals is a true response or is due to an error in experimental design. Page 25 of 87 Guideline No./ Study Type MRID No. ( year)/ Classification / Doses Results mg/ kg/ day E2.

4.2 FQPA Hazard Considerations 4.2.1 Adequacy of the Toxicity Database The toxicology database for benthiavalicarb is complete and adequate for FQPA assessment. Acceptable/ guideline studies for developmental toxicity in rats and rabbits and a 2 generation reproduction study in rats were available for consideration during endpoint selection.

4.2.2 Evidence of Neurotoxicity Neurotoxicity was not observed in either the acute or subchronic neurotoxicity studies up to the limit dose of 2000 mg/ kg/ day. Furthermore, no evidence of neurotoxicity was seen throughout the entire toxicology database.

4.2.3 Developmental Toxicity Studies Developmental effects were not seen in either the developmental rat or rabbit studies. There were no effects on survival, body weights, incidences of abortions, premature deliveries, or litter resorptions. There were no effects on external, visceral, or skeletal malformations, variations, or retardations.

The maternal NOAEL ( 100 mg/ kg/ day) in rats was based upon liver effects ( increased absolute and relative liver weights and increased incidence of enlarged liver) observed at the LOAEL ( 1000 mg/ kg/ day). Microscopic examination of the livers was not performed in the developmental study; however, when compared to the liver lesions observed in the subchronic rat study, the reviewer noted that the effects on the liver were similar to the liver changes in other studies and were considered adverse.

The maternal NOAEL ( 20 mg/ kg/ day) in the rabbit developmental study was based upon increased absolute ( 10%) and relative ( 11%) liver weights at the LOAEL of 40 mg/ kg/ day.

4.2.4 Reproductive Toxicity Study No adverse effects ( maternal, reproductive, or developmental) were seen in the 2 generation reproduction study in rats. Increased liver weights, slight hepatocyte hypertrophy, and two incidences of enlarged liver were observed in the high dose group. However, in the absence of other corroborating pathological indications of toxicity, the effect on liver size was regarded as an adaptive response.

4.2.5 Additional Information from Literature Sources Page 26 of 87 An open literature review for benthiavalicarb has not been conducted. However, benthiavalicarb is registered for use in Europe, and the European monograph is available. The monograph was consulted and used as supplementary information.

4.2.6 Pre and/ or Post natal Toxicity Developmental studies in two species ( both rat and rabbit) and a 2 generation reproduction study in rats did not show any evidence of developmental or reproductive toxicity.

<IP_ADDRESS> Determination of Susceptibility The toxicology database is complete and adequate to assess questions of increased qualitative and/ or quantitative susceptibility in infants and children. Evidence suggests that there is no concern for fetuses exposed to benthiavalicarb in utero or post natally. Maternal toxicity is observed in both the rat and rabbit developmental studies in the absence of developmental effects to the fetuses. No adverse effects were observed in the 2 generation reproduction study.

<IP_ADDRESS> Degree of Concern Analysis and Residual Uncertainties for Pre and/ or Post natal Susceptibility There is no concern for increased risk to developing fetuses resulting from exposure in utero and/ or post natally. Therefore, the FQPA factor has been reduced to 1x.

4.3 Recommendation for a Developmental Neurotoxicity Study The registrant submitted acceptable acute and subchronic neurotoxicity studies. There was no evidence of neurotoxicity up to the limit dose of 2000 mg/ kg/ day. Because of the absence of neurotoxicity throughout the entire toxicology database and the absence of adverse developmental and reproductive effects, a developmental neurotoxicity study is not necessary at this time. Page 27 of 87 4.4 Hazard Identification and Toxicity Endpoint Selection 4.4.1 Acute Reference Dose ( aRfD)  All Populations Because of the absence of toxic effects attributable to a single dose, no acute endpoint has been selected for risk assessment.

4.4.2 Chronic Reference Dose ( cRfD)

The chronic reference dose was selected from the chronic rat study based upon nephrotoxicity and hepatotoxicity. In particular, hepatotoxicity is prevalent throughout the entire toxicology database and is the main effect of concern. Furthermore, the NOAEL of 9.9 mg/ kg/ day is protective of tumor effects seen in both rats and mice at doses greater than 250 mg/ kg/ day. The standard 10x intraspecies variability and 10x interspecies extrapolation safety factors apply. Therefore, the cRfD = 0.099 mg/ kg/ day.

4.4.3 Non Dietary Exposure for Handlers The registrant is only seeking import tolerances for benthiavalicarb. As a result, the only relevant route of exposure is through food. Occupational and residential exposures do not need to be assessed at this time.

4.4.4 Margins of Exposure The level of concern for risk assessment is margins of exposure that are less than 100, based on the standard 10x interspecies and 10x intraspecies extrapolation factors.

4.4.5 Recommendation for Aggregate Exposure Risk Assessments The registrant is seeking import tolerances on grapes and tomatoes. Therefore, the risk assessment only includes dietary exposure to benthiavalicarb via imported grapes and tomatoes.

4.4.6 Classification of Carcinogenic Potential In accordance with EPA's Final Guidelines for Carcinogen Risk Assessment ( March, 2005), the CARC classified benthiavalicarb as " likely to be carcinogenic to humans," based on the presence of malignant uterine tumors in rats, liver tumors in both sexes of mice ( with some supporting evidence found in liver tumors in male rats), and thyroid follicular cell tumors in male mice, at doses adequate to assess carcinogenicity. Benthiavalicarb is not mutagenic.

4.5 Special FQPA Safety Factor The toxicology database is complete and adequate for FQPA analysis. There is no concern for developmental and/ or reproductive effects, increased susceptibility, or neurotoxicity. Therefore, HED has reduced the FQPA safety factor to 1x. The endpoints selected for risk assessment are based on the effects of concern ( hepatotoxicity), and are protective of tumor types observed in the chronic/ carcinogenicity studies. There is little residual uncertainty with regard to the toxicology Page 28 of 87 database.

Table 4.5. Summary of Toxicological Doses and Endpoints for Use in Human Health Risk Assessments Exposure Scenario Dose Used in Risk Assessment, UF Special FQPA SF* and Level of Concern for Risk Assessment Study and Toxicological Effects Acute Dietary ALL populations including infants and children when applicable NOAEL= N/ S UF = N/ A Acute RfD = N/ A FQPA SF = N/ A None of the study endpoints were the result of one or two exposures to benthiavalicarb; therefore an acute endpoint was not selected for dietary exposure.

Chronic Dietary all populations NOAEL= 9.9 mg/ kg/ day UF = 100 Chronic RfD = 0.099 mg/ kg/ day FQPA SF = 1X cPAD = 0.099 mg/ kg/ day Chronic Oral Toxicity in Rats LOAEL = 249.6 mg/ kg/ day based on nephrotoxicity and hepatotoxicity.

Cancer ( oral, dermal, inhalation) Likely to be carcinogenic to humans Q1* = 6.2795x10 2 Based on increases in male mouse liver combined adenomas and/ or carcinomas and/ or blastomas.

UF = uncertainty factor, FQPA SF = Special FQPA safety factor, NOAEL = no observed adverse effect level, LOAEL = lowest observed adverse effect level, PAD = population adjusted dose ( a = acute, c = chronic) RfD = reference dose, MOE = margin of exposure, LOC = level of concern, NA = Not Applicable * Refer to Section 4.5 4.6 Endocrine disruption EPA is required under the FFDCA, as amended by FQPA, to develop a screening program to determine whether certain substances ( including all pesticide active and other ingredients) " may have an effect in humans that is similar to an effect produced by a naturally occurring estrogen, or other such endocrine effects as the Administrator may designate." Following recommendations of its Endocrine Disruptor and Testing Advisory Committee ( EDSTAC), EPA determined that there was a scientific basis for including, as part of the program, the androgen and thyroid hormone systems, in addition to the estrogen hormone system. EPA also adopted EDSTAC's recommendation that the Program include evaluations of potential effects in wildlife. For pesticide chemicals, EPA will use FIFRA and, to the extent that effects in wildlife may help determine whether a substance may have an effect in humans, FFDCA authority, to require the wildlife evaluations. As the science develops and resources allow, screening of additional hormone Page 29 of 87 systems may be added to the Endocrine Disruptor Screening Program ( EDSP). In the available toxicity studies on benthiavalicarb, there was no estrogen, androgen, and/ or thyroid mediated toxicity. When additional appropriate screening and/ or testing protocols being considered under the Agency's EDSP have been developed, benthiavalicarb may be subjected to further screening and/ or testing to better characterize effects related to endocrine disruption.

5.0 Public Health Data As benthiavalicarb will not be used in the U. S., public health data are not relevant to this tolerance petition.

6.0 Exposure Characterization/ Assessment 6.1 Dietary Exposure/ Risk Pathway 6.1.1 Residue Profile A discussion of benthiavalicarb residue chemistry is given in a memorandum prepared for these tolerance requests ( D322934, D. Dotson, 3/ 2/ 2006). The nature of the residue has been adequately delineated for the proposed uses based on the acceptable grape and tomato metabolism studies. The available potato metabolism study is also acceptable pending submission of supporting storage stability data ( 21 months). In grapes and tomatoes, the metabolism of benthiavalicarb was minimal with parent being the only major residue in fruit and foliage ( 55 95% TRR). Parent was also the principal 14C residue in potato foliage ( 88 90% TRR), but other minor metabolites were identified. In potatoes, the metabolism of benthiavalicarb occurs primarily via direct hydroxylation of the phenyl ring or defluorination and hydroxylation of the phenyl ring. Secondary metabolism occurs via conjugation of sugars with hydroxy groups. Chiral analysis of potato foliage extracts also indicates that there was no isomeric conversion of the active R L isomer to other possible isomers ( S L, R D, or S D). Based on the results of the metabolism studies, the residues of concern in grapes and tomatoes for both tolerance expression and risk assessment are the combined residues of the R L and S L isomers of benthiavalicarb. The nature of the residue in livestock was not determined, and is not required, as there are no livestock feed items associated with grapes and tomatoes.

An adequate GC/ NPD Method ( RCC No. 665943) is available for collecting data on residues of both the R L and S L isomers of benthiavalicarb in/ on grape and tomato matrices. For this method, residues are extracted with acetone, partitioned into hexane, purified using an amino SPE cartridge, and determined by GC/ NPD. The lower limit of method validation ( LLMV) is 0.01 ppm for each analyte in grape and tomato matrices. The reported limit of detection ( LOD) in grapes is 0.001 ppm. The reported LOD in grape processed commodities, tomatoes, and tomato processed commodities is 0.002 ppm. Although the submitted reports state that the LOQ was 0.01 ppm, an inspection of the chromatograms indicates that the LOQ would probably be in the range of 0.002 0.003 ppm for the various grape and tomato commodities. In many cases residue values were reported as being between 0.001 and 0.01 ppm ( i. e., actual values were reported as 0.002 ppm, 0.003 ppm, etc.). The reporting of such values is further evidence that the LOQ is Page 30 of 87 indeed below 0.01 ppm. The method was adequately validated by the petitioner, and is being proposed as the tolerance enforcement method. The method was successfully validated in an independent laboratory method validation ( ILV) trial, and radiovalidation data are not required, given the similarity between the extraction procedures in the method and the metabolism studies. However, because the method is not highly specific, the petitioner should either develop a confirmatory method for the analytes of interest or conduct an interference study. The method was forwarded to BEAD/ ACB for a petition method validation ( PMV) trial. The method was forwarded to the Analytical Chemistry Branch ( ACB) of the Biological and Economic Assessment Division ( BEAD) for a petition method validation ( PMV) trial. ACB concluded that the method does not need to be laboratory validated by EPA, and appears to be suitable for tolerance enforcement.

Adequate storage stability data are available indicating that benthiavalicarb is stable under frozen (  18 ° C) conditions for at least 13 months in grapes and 8 months in tomatoes. These data support the maximum storage intervals ( 7 8 months) from the grape and tomato field trials.

As there are no significant animal feed items associated with the proposed use on grapes and tomatoes, livestock feeding studies are not required for this petition. Tolerances do not need to be established on animal commodities.

The available field trial data are adequate and will support the proposed use patterns on grapes and tomatoes. The number and geographic distribution of the field trials are adequate. For grapes, a total of 34 field tests were conducted throughout the EU, in which grapes were treated at 1x the maximum proposed rate and harvested at the minimum PHI of approximately 28 days. Maximum combined residues were < 0.27 ppm in/ on grapes, and the HAFT residue value was 0.22 ppm. For tomatoes, 12 tests were conducted at 1x the maximum rate throughout the EU on both greenhouse and outdoor field grown tomatoes. Nine of these 12 field trials were performed on greenhouse grown tomatoes and three were performed on outdoor grown tomatoes. Maximum combined residues were 0.27 ppm in/ on greenhouse tomatoes 3 days after treatment ( DAT) and < 0.015 ppm in/ on field grown tomatoes ( 7 DAT). The recommended tolerances for grapes and tomatoes were generated using HED's statistical tolerance generator for NAFTAharmonized tolerances. The recommended tolerance for grapes is 0.25 ppm, even though the highest field trial value is 0.27 ppm. The recommended tolerance for tomatoes is 0.45 ppm.

Overall, 22 grape field trials were performed in which samples were taken at 1 week intervals. The results of these decline studies can be found in Table A. 6. The samples were taken at 0, 7, 14, 21, and 28 DAT. In four of those trials, samples were not taken at 28 days. Average residues declined as follows: 0.088 ppm, 0.070 ppm, 0.055 ppm, 0.049 ppm, and 0.046 ppm at 0, 7, 14, 21, and 28 days, respectively. Residues declined by 0.018 ppm, 0.015 ppm, 0.06 ppm, and 0.03 ppm over the four intervals. In effect, residues declined with the greatest decreases occurring in the early stages of the field trials.

The available processing data are adequate, pending submission of supporting storage stability data for processed matrices. The average processing factors for the combined residues were 3.7x in raisins, 0.47x in juice, and 0.97x in wine. Based on the 3.7x processing factor for raisins and the HAFT residue value of 0.22 ppm for grapes, the maximum expected residues in raisins would be 0.81 ppm, which is above the recommended grape tolerance. Therefore, a separate tolerance is Page 31 of 87 required for raisins. HED recommends in favor of the proposed tolerance of 1.0 ppm for grape, raisin. Storage stability data were not submitted for processed grape and tomato commodities. These data are not required for the current tolerance requests because acceptable storage stability data were submitted for grapes and tomatoes. Residues were stable in grapes for 13 months and in tomatoes for 8 months. In the tomato storage stability studies, tomatoes were homogenized prior to storage. Homogenized tomatoes are comparable in both composition and texture to processed tomato commodities. The matrix is an acidic one which would very likely maximize residue decay. As a result, the tomato data are considered adequate to demonstrate storage stability in processed grape commodities as well.

In the tomato processing studies, the average processing factors from tests having quantifiable residues were 0.49x in juice, 1.2x in puree, and 1.4x in ketchup. As residue data were not available for paste, the 5.5x theoretical concentration factor based on loss of water was used for paste. The HAFT residue value of 0.015 ppm from the tests on field grown tomatoes was used to assess the need for tolerances on processed tomato fractions as greenhouse tomatoes are not utilized for processing. Based on the above processing factors and the highest field trial value of 0.015 ppm, the maximum expected residues in puree, ketchup, juice, and paste would be 0.018, 0.021, 0.0074 ppm, and 0.083 ppm, respectively, which are all below the recommended tomato tolerance of 0.45 ppm. Therefore, separate tolerances are not required for processed tomato fractions.

As the proposed uses are for grape and tomatoes grown in the EU, confined and field rotational crops studies are not required for the current petition.

6.1.2 Acute and Chronic Dietary Exposure and Risk A chronic dietary risk assessment was conducted using the Dietary Exposure Evaluation Model ( DEEM FCID, Version 2.03). The dietary exposure analysis and its results are discussed in Memo, D322935, D. Dotson, 3/ 2/ 2006. The DEEM FCID Model uses food consumption data from the USDA's Continuing Surveys of Food Intakes by Individuals ( CSFII) from 1994 1996 and a supplemental children's survey conducted in 1998. The result of the chronic dietary exposure assessment for the general U. S. population was used along with the cancer Q1* value to determine the cancer risk to the U. S. population.

In order to determine the percent crop imported for individual grape and tomato commodities, data are needed for the following three quantities: ( 1) domestic production, ( 2) the quantity exported, and ( 3) the quantity imported. These data were not available for all commodities in the analysis. For grapes, these data were available for grape juice and raisins only. Percent imported data for grapes and wine are directly available from the USDA Economic Research Service ( ERS), however. For unprocessed tomatoes, the three quantities listed above are available. Data were not used for the individual processed tomato commodities because a significant amount of processing occurs after importation. Data are available for domestic tomato production utilized for processing and processed tomato imports. The import data include canned tomatoes, a significant fraction of which is utilized for processing in the U. S. Dividing the imported by the total yields a value of 5.84% imported. The USDA ERS, Vegetable and Melons Outlook ( VGS 308) of April 21, 2005 states the following: " According to ERS estimates, nearly 6 percent of the Page 32 of 87 tomato products consumed by Americans in 2004 were imported." The value of 5.84% was used for paste, puree, and juice. A value of 100% was used for dried tomatoes.

The data for grape commodities were provided by BEAD ( electronic communication, D. Donaldson, 2/ 1/ 2006). USDA ERS data for 2002 and 2003 were used. USDA ERS data from 2004 were used for tomatoes. The following percent crop imported values were used in the analysis: tomatoes ( 36.3%), processed tomato commodities ( 5.84%), grapes ( 45%), grape juice ( 43.9%), wine ( 23%), and raisins ( 10.7%).

Chronic Dietary Exposure Results and Characterization The chronic dietary exposure analysis is based on average field trial values and estimates of percent crop imported. Empirical processing factors were used for most processed commodities. When empirical factors were not available, DEEM ( Version 7.81) default processing factors were used. The general U. S. population and all population subgroups had risk estimates that were below HED's level of concern ( i. e., 100% of the cPAD). In fact, the U. S. population and all population subgroups had risk estimates that were less than 1% of the cPAD. For the chronic assessment, all population subgroups would have had risk estimates that were below HED's level of concern if no refinements had been made to the analysis, i. e., if tolerance level residues and 100% crop imported assumptions had been used. However, it was necessary to refine the inputs for the cancer assessment. For simplicity, the same refinements were used for the chronic analysis.

Cancer Dietary Exposure Results and Characterization As stated above, the CARC determined that benthiavalicarb is likely to be carcinogenic to humans and assigned it a Q1* value of 0.063 ( mg/ kg/ day) 1. In an unrefined cancer dietary exposure analysis, the estimated cancer risk to the general U. S. population exceeds HED's level of concern ( i. e., 1.0 x 10 6 or one in one million). As a result, refinements were made to the cancer dietary exposure analysis. The refinements made are the same as those made for the chronic analysis discussed above: the use of average field trial values and estimates of percent crop imported. With the use of these refinements, the estimated lifetime cancer risk to the U. S. population is 1.6 x 10 6. Refinements were made to the analysis; however, it is still considered to be conservative. First, average field trial values from field trials almost always exceed the expected residue levels found on crops at the time of consumption. When field trials are performed, the maximum allowable application rate is used and crops are harvested at the minimum PHI. Samples are stored frozen until analysis to ensure minimal degradation of residues. In actual practice, however, growers will not usually use the maximum application rates for economic reasons. In addition, most crops are not harvested and immediately stored frozen. From previous dietary exposure analyses, HED has noted that monitoring data are often one to two orders of magnitude lower than field trial data. The second reason this analysis is considered to be conservative is that the registrant is planning on marketing benthiavalicarb in Europe only. In the dietary analysis, percent crop imported estimates were used for all imports, not just those from Europe. The third reason this analysis is considered to be conservative is that HED assumed that 100% of the imported crop will be treated. The actual percent crop treated will very likely be considerably lower than that. For these reasons, HED is confident that this analysis overestimates risk to the general U. S. population. The cancer risk to the general U. S. population does not exceed HED's Page 33 of 87 level of concern.

Table 6.1. Summary of Dietary Exposure and Risk for Benthiavalicarb DEEM: Chronic Analysis Population Subgroup Acute Analysis Dietary Exposure ( mg/ kg/ day) % cPAD Cancer Analysis General U. S. Population 0.000026 < 0.1 1.6 x 10 6 All Infants (< 1 year old) Not Applicable: No Acute Dietary Endpoint 0.000017 < 0.1 Children 1 2 years old 0.000071 0.1 Children 3 5 years old 0.000053 0.1 N/ A: Cancer risk is determined for the general U. S. population only Children 6 12 years old 0.000030 < 0.1 Youth 13 19 years old 0.000019 < 0.1 Adults 20 49 years old 0.000022 < 0.1 Adults 50+ years old 0.000023 < 0.1 Females 13 49 years old 0.000021 < 0.1 The commodities that made the greatest contribution to the risk estimates are tomatoes, grape juice, grapes, wine, and raisins. For the general U. S. population, these commodities contribute a total of 98% of the entire cancer risk. The individual contributions are: tomatoes ( 61%), grape juice ( 14%), grapes ( 12%), wine ( 8%), and raisins ( 3%).

HED is confident that this analysis does not underestimate risk to the general U. S. population. HED is also confident that average field trial values and percent crop imported values used in the assessment will not be exceeded in the future. For the reasons stated above, the field trial values considerably overestimate residues found on food at the time of consumption. It is unlikely that residue levels in imported grape and tomato commodities at the time of consumption would ever exceed the average field trial values. As far as percent crop imported values being exceeded in the future, this situation is unlikely as well. As stated above, the conservative assumptions were made that 100% of the grape and tomato commodities imported from all over the world would be treated. It's very unlikely that the quantities of imports actually treated will ever increase to those levels. Data for grape commodities were provided by BEAD. The economist who provided the data stated that it was unlikely the estimates of import shares would be exceeded in the near future. The data for grape commodities provided to HED were five year averages, and were compared to the annual data points of each data source, when available. Import share data do vary from year to year, but crop production patterns and consumption and preferences are more stable and change more slowly. For tomatoes, the worldwide imports changed as follows from 1998 to 2004: 37%, 31%, 30%, 33%, 32%, 37%, and 36%. Clearly, no increase occurred over that time period. For processed tomato commodities, the worldwide imports changed as follows from 2000 to 2004: 2.9%, 5.6%, 7.1%, 5.4%, and 5.8%. Between 2001 and 2004 there was no significant increase in processed tomato commodity imports. Page 34 of 87 6.2 Water Exposure/ Risk Pathway The tolerances being granted in conjunction with these requests are for imported commodities only. As a result, residues in water are not being considered in this assessment.

6.3 Residential ( Non Occupational) Exposure/ Risk Pathway There are no residential uses for benthiavalicarb and the food uses are for imported commodities. As a result, residential uses ( including home uses, recreational uses, spray drift, etc.) are not relevant to this assessment.

7.0 Aggregate Risk Assessments and Risk Characterization 7.1 Acute Aggregate Risk None of the toxicity study endpoints were the result of a single oral exposure to benthiavalicarb. As a result, no acute dietary endpoint was identified, and an acute risk assessment is not needed.

7.2 Short, Intermediate, and Long Term Aggregate Risk As the current petition is for import tolerances on grapes and tomatoes only, there are no residential uses, and residues in drinking water are not of concern. As a result, short and intermediate term aggregate risk assessments are not needed. The chronic aggregate risk assessment is equivalent to the chronic dietary exposure analysis. See Section 6.1.2. for a discussion of the dietary exposure analysis. The general U. S. population and all population subgroups had risk estimates that were well below HED's level of concern.

7.3 Cancer Risk The cancer aggregate risk assessment is equivalent to the cancer dietary exposure analysis. See Section 6.1.2., above for a discussion of this analysis. With the use of some refinements, the lifetime cancer risk to the U. S. population is 1.6 x 10 6. HED believes this risk estimate is still an overestimate, however, and that the cancer risk is not of concern.

8.0 Cumulative Risk Characterization/ Assessment Unlike other pesticides for which EPA has followed a cumulative risk approach based on a common mechanism of toxicity, EPA has not made a common mechanism of toxicity finding as to benthiavalicarb and any other substances, and benthiavalicarb 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 benthiavalicarb has a common mechanism of toxicity with other substances. Although benthiavalicarb is a carbamate compound, it is not a member of the class of insecticides known as the N methyl carbamates for which the Agency is presently conducting a cumulative risk assessment. The substituents on the benthiavalicarb nitrogen atom are much larger than the methyl group in the insecticides. While the N methyl carbamates are Page 35 of 87 neurotoxicants based on their ability to inhibit the enzyme cholinesterase, there is no evidence of neurotoxicity or neuropathology in the hazard database for benthiavalicarb. Benthiavalicarb is also not a member of the thiocarbamate herbicides or dithiocarbamate fungicides.

For information regarding EPA's efforts to determine which chemicals have a common mechanism of toxicity and to evaluate the cumulative effects of such chemicals, see the policy statements released by EPA's Office of Pesticide Programs concerning common mechanism determinations and procedures for cumulating effects from substances found to have a common mechanism on EPA's website at http:// www. epa. gov/ pesticides/ cumulative/.

9.0 Occupational Exposure/ Risk Pathway As benthiavalicarb will be used outside the U. S. to treat grapes and tomatoes, there will be no occupational risk to U. S. workers. As a result, an occupational assessment is not relevant to this tolerance petition.

10.0 Data Needs and Label Requirements 10.1 Toxicology The toxicology database is complete and adequate for risk assessment. There are no further toxicology requirements at this time.

10.2 Residue Chemistry No major deficiencies were noted in the subject petition that would preclude establishing tolerances for benthiavalicarb on grape and tomato commodities. However, before any further tolerances are granted, the petitioner should either develop a confirmatory analytical method for the analytes of interest or conduct an interference study.

There are no residue chemistry issues that would preclude establishing tolerances of 0.25 ppm for grape, 0.45 ppm for tomato, and 1.0 ppm for grape, raisin. The recommended tolerances for grape and tomato were generated using HED's statistical tolerance generator for NAFTAharmonized tolerances. The residues to be included in the tolerance expression are the combined R L and S L isomers of benthiavalicarb. The petitioner has recommended that only the R L isomer be included in the tolerance expression because the S L isomer is a minor impurity in the commercial formulations and the potato metabolism study indicates that the active R L isomer is not converted to the S L isomer. In addition, the residue definition proposed for the EU MRLs includes only the R L isomer. The registrant has proposed EU MRLs of 0.2 ppm for grapes and 0.3 ppm for tomatoes. It's possible that if EU MRLs are established, the residue definition might be benthiavalicarb isopropyl only, and no distinction will be made as to the isomer. HED concludes that the S L isomer should be included in the tolerance expression, however. The results of the field trials indicate that the residue levels of the S L isomer are often non negligible compared to the levels of the R L isomer. In the grape field trials several samples had S L isomer levels that were 10 20% of the R L isomer levels. A small number of samples had S L: R L ratios that exceeded 20%. In the tomato field trials, the S L residue levels were generally non Page 36 of 87 detectable. In a few cases, however, the S L isomer was present at 6 7% of the level of the R L isomer. The technical material contains 93% R L isomer and 3.4% S L isomer. The analytical method determines both isomers separately.

10.3 Occupational and Residential Exposure There are no data needs or label requirements.

11.0 Summary of Proposed and Recommended Tolerances Table 11.1 gives the proposed tolerances, the recommended tolerances, and the correct commodity definitions for the commodities associated with this risk assessment. The registrant did not propose a tolerance for wine and none is needed. Residues do not concentrate in wine; therefore, the 0.25 ppm tolerance recommended for grape is adequate to cover wine.

Table 11.1 Tolerance Summary for Benthiavalicarb Isopropyl ( PP# 3E6545)

Commodity Proposed Tolerance ( ppm) Recommended Tolerance ( ppm) Correct commodity definition and comments Grapes 0.5 0.25 Grape The available data are adequate. Maximum residues in/ on grapes were 0.27 ppm.

Raisins 1.0 1.0 Grape, Raisin The available data are adequate. Based on an average processing factor of 3.7x for raisins and the HAFT residue value in grapes of 0.22 ppm, maximum residues in raisins would be 0.81 ppm.

Tomatoes 0.5 0.45 Tomato The available data are adequate. Maximum residues were 0.26 ppm in/ on greenhouse grown tomatoes and < 0.015 ppm in/ on field grown tomatoes.

Tomato Paste 1.5 None Tomato, Paste The available data are adequate. Average processing factors were 1.2x and 1.4x for tomato puree and ketchup, respectively, and were estimated to be 5.5x for paste ( the maximum theoretical concentration factor). Based on the highest field trial residue value of 0.015 ppm for field grown tomatoes, the maximum expected residues in tomato paste would be 0.083 ppm. Page 37 of 87 References:

Benthiavalicarb Isopropyl: Report of the Cancer Assessment Review Committee, TXR No. 0053803, J. Kidwell, 10/ 18/ 2005 Benthiavalicarb: Quantitative Risk Assessment ( Q1*) Based on Fischer F 344/ DuCrj Rat and SLC: B6C3F1( C57BL/ 6xC3H) SPF Mouse Dietary Studies With ¾ ' s Interspecies Scaling Factor, TXR No. 0053755, Lori Brunsman, 9/ 26/ 2005 Benthiavalicarb Isopropyl. PP# 3E6545. Request for Tolerances on Imported Grapes and Tomatoes. Summary of Analytical Chemistry and Residue Data, D322934, D. Dotson, 3/ 2/ 2006.

Benthiavalicarb Isopropyl: Chronic and Cancer Dietary Exposure Assessments for the Section 3 Registration Action on Imported Grapes and Tomatoes, D322935, D. Dotson, 3/ 2/ 2006. Page 38 of 87 Appendices 1.0 TOXICOLOGY DATA REQUIREMENTS The requirements ( 40 CFR 158.340) for import tolerance on food crops for benthiavalicarb are in Table 1. Use of the new guideline numbers does not imply that the new ( 1998) guideline protocols were used.

Table A. 1. Toxicology Data Requirements Technical Test Required Satisfied 870.1100 Acute Oral Toxicity 870.1200 Acute Dermal Toxicity 870.1300 Acute Inhalation Toxicity 870.2400 Primary Eye Irritation 870.2500 Primary Dermal Irritation 870.2600 Dermal Sensitization yes no no no no no yes

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

870.3700a Developmental Toxicity ( rodent) 870.3700b Developmental Toxicity ( nonrodent) 870.3800 Reproduction yes yes yes yes yes yes 870.4100a Chronic Toxicity ( rodent) 870.4100b Chronic Toxicity ( nonrodent) 870.4200a Oncogenicity ( rat) 870.4200b Oncogenicity ( mouse) 870.4300 Chronic/ Oncogenicity yes yes yes yes yes yes yes yes yes yes 870.5100 Mutagenicity_ Gene Mutation  bacterial 870.5300 Mutagenicity_ Gene Mutation  mammalian 870.5xxx Mutagenicity_ Structural Chromosomal Aberrations 870.5xxx Mutagenicity_ Other Genotoxic Effects yes yes yes yes yes yes yes yes 870.6100a Acute Delayed Neurotoxicity. ( hen) 870.6100b 90 Day Neurotoxicity ( hen) 870.6200a Acute Neurotoxicity. Screening Battery ( rat) 870.6200b 90 Day Neurotoxicity. Screening Battery ( rat) 870.6300 Develop. Neurotoxicity no no no no no   yes yes

870.7485 General Metabolism 870.7600 Dermal Penetration yes no yes

Special Studies for Ocular Effects Acute Oral ( rat) Subchronic Oral ( rat) Six month Oral ( dog) no no no no     Page 39 of 87 2.0 TOXICOLOGY STUDIES 2.1. Subchronic Toxicity Studies 2.1.1. Inoue, H. ( 1998) KIF 230 Technical: subchronic toxicity study in rats by dietary administration for 13 weeks followed by a 4 week recovery study. Biosafety Research Center, Foods, Drugs and Pesticides ( An Pyo Center) Shizuoka, Japan. Laboratory Study No.: 3386, December 18, 1998. MRID 45835004. Unpublished.

EXECUTIVE SUMMARY  In a subchronic oral toxicity study ( MRID 45835004), Benthiavalicarb ( KIF 230; 87.7% a. i., Lot #: G51 02 152) was administered to 10 Fischer rats/ sex/ group in the diet at dose levels of 0, 50, 200, 5000, or 20,000 ppm ( equivalent to 0/ 0, 3.5/ 3.9, 14.1/ 15.3, 353/ 379, and 1444/ 1552 mg/ kg/ day [ M/ F]) for 13 weeks. An additional 10 rats/ sex/ group were concurrently administered the 0, 5000, and 20,000 ppm diets for 13 weeks and were retained for a recovery period of 4 weeks.

There were no adverse effects of treatment on survival, body weights, body weight gains, food consumption, food efficiency, ophthalmoscopy, or hematology.

Absolute and relative ( to body) liver weights were increased ( p  0.01) in both sexes at   5000 ppm at Week 13 and remained increased ( p  0.01) in the 20,000 ppm females at Week 17. Hepatocyte hypertrophy was observed in both sexes at 5000 ppm ( 3/ 10 treated vs 0/ 10 controls) and 20,000 ppm ( 10/ 10 treated). Additionally in both sexes at 20,000 ppm, black and enlarged liver was observed macroscopically. These findings were not observed at the end of the recovery period.

At Week 13, the following clinical chemistry parameters were dose dependently increased ( p  0.05) over controls and corroborated the toxicity to the liver: ( i) total cholesterol and GGT at   5000 ppm in both sexes; ( ii) free cholesterol and phospholipids in the 20,000 ppm males and   5000 ppm females; ( iii) total protein in the   5000 ppm males and 20,000 ppm females; and ( iv) increased albumin in both sexes at 20,000 ppm. At the end of the recovery period, increased ( p  0.05) GGT and albumin were still observed in both sexes at   5000 ppm, and a minor increase in total protein ( p  0.05) was noted in the 20,000 ppm females, although to a lesser extent than in the main study animals at Week 13. In general, serum protein electrophoresis demonstrated increases in globulin that exceeded increases in albumin, resulting in a decreased A/ G ratio that may support an effect on the liver at   5000 ppm.

The LOAEL for this study is 5000 ppm ( equivalent to 352.6/ 378.5 mg/ kg/ day [ M/ F]) based on hepatocyte hypertrophy in both sexes and increases in: absolute and relative to body liver weights, total cholesterol, and GGT in both sexes; free cholesterol and phospholipids in the females; and total protein in the males. The NOAEL is 200 ppm ( equivalent to 14.1/ 15.3 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 rat.

2.1.2. Inoue, H. ( 1998) KIF 230 Technical: Preliminary oncogenicity study in mice by dietary administration for 13 weeks. Biosafety Research Center, Shizuoka, Japan. Laboratory Report Page 40 of 87 No.: 3385, November 27, 1997. MRID 45835007. Unpublished.

EXECUTIVE SUMMARY  In a subchronic oral toxicity study ( MRID 45835007), KIF 230 ( benthiavalicarb; 87.7% a. i., Lot #: G51 02 152) was administered to 10 B6C3F1 mice/ sex/ dose in the diet at doses of 0, 50, 200, 7000, or 20,000 ppm ( equivalent to 0/ 0, 8.4/ 11.3, 33.0/ 45.2, 1293/ 1620, or 4031/ 4946 mg/ kg/ day [ M/ F]) for up to 90 days.

Urinalysis, clinical chemistry, neurological evaluations, and ophthalmoscopic examinations were not performed. No compound related effects on mortality, clinical signs, or hematology were observed at any dose in either sex.

Body weights were decreased ( p  0.05) in the 7000 ppm males at Weeks 6 13, and at 20,000 ppm in the males at Weeks 3 13 and in the females at Weeks 4 10. Overall ( Weeks 0 13) body weight gains were decreased ( p  0.01) in the   7000 ppm males and in the 20,000 ppm females ( not statistically significant). Food efficiency was decreased ( p  0.05) in the 7000 ppm males at Weeks 4, 7, 9 10, and 13. At 20,000 ppm, food consumption was sporadically increased ( p  0.05) in the males and females, and overall food consumption was slightly increased ( p  0.01) in the males. Food efficiency was decreased throughout the study in the males, with statistical significance ( p  0.05) achieved at Weeks 3 5, 9, and 13, and during Weeks 1 5 in the females, with statistical significance ( p  0.01) being achieved at Week 5. Overall food efficiency was decreased ( p  0.05) in the   7000 ppm males and in the 20,000 ppm females.

The target organ was the liver. At   7000 ppm, absolute liver weights were increased ( p  0.01) in both sexes, and relative ( to body) liver weights were also increased ( p  0.01) in these animals. Gross lesions observed at   7000 ppm included enlarged liver in all animals and black colored liver in 7 10 females. Additionally at 20,000 ppm, black colored liver was noted in all males and a white patch/ zone in the liver was noted in 6/ 10 females. Histopathological effects noted in the liver included: ( i) necrosis in the   7000 ppm males and females; ( ii) slight bile duct hyperplasia in the 20,000 ppm males and females; ( iii) fatty change in the 20,000 ppm males; ( iv) slight to moderate anisonucleosis in the   7000 ppm males; ( v) multinucleated giant cell formation in the 20,000 ppm males; and ( vi) slight to marked hepatocellular hypertrophy in the   7000 ppm males and females. Electron microscopy of the liver samples from the 7000 and 20,000 ppm groups revealed increases in rough endoplasmic reticulum associated with the hepatocellular hypertrophy.

Additional findings included brown colored thyroid in all 20,000 ppm males.

No treatment related effects were observed at   200 ppm in either sex.

The LOAEL is 7000 ppm ( equivalent to 1293/ 1620 mg/ kg/ day [ M/ F]; greater than the limit dose) based on decreased body weight, body weight gain, and food efficiency in males, increased absolute and relative liver weight, enlarged liver, black colored liver ( females only), and histopathological liver effects ( necrosis, hypertrophy, and anisonucleosis [ males only]). The NOAEL is 200 ppm ( equivalent to 33.0/ 45.2 mg/ kg/ day [ M/ F]). This study is classified acceptable/ guideline and satisfies the guideline requirement ( OPPTS 870.3100; OECD 408) for a 90 day oral toxicity study in the mouse. Page 41 of 87 2.1.3. Subchronic Toxicity Study in Dogs EXECUTIVE SUMMARY: In a subchronic oral toxicity study ( MRID 45835006), Benthiavalicarb ( KIF 230); ( 88.8% a. i., Lot/ Batch #: G51 08 158) was administered daily to 4 beagle dogs/ sex/ group in gelatin capsules at dose levels of 0, 40, 200, or 1000 mg/ kg/ day for 13 weeks.

At 1000 mg/ kg/ day, liquid stool was observed in three males and one female at 1000 mg/ kg/ day. Hematocrit, hemoglobin, and red blood cell counts were decreased ( p  0.05), while the percentage of reticulocytes and the concentration of serum total bilirubin were increased in both sexes. These changes were indicative of a hemolytic anemia. Moderate deposits of hemosiderin pigment were also observed in the spleen and Kupffer cells of the liver in all animals at this dose, indicative of secondary effects of a hemolytic anemia. Serum gamma glutamyl transferase, alkaline phosphatase, and platelet counts were increased ( p  0.05) in both sexes. Serum albumin in both sexes and creatine kinase and albumin/ globulin ratio in the females were decreased ( p  0.05). In females, absolute and relative ( to body) thymus weights were decreased ( p  0.01). Absolute and relative ( to body) liver weights were increased ( p  0.01) and enlarged liver was grossly observed in 4/ 4 males and 3/ 4 females at this dose compared to 0/ 4 in controls. Slight diffuse lobular hepatocyte hypertrophy was also observed in all animals at this dose.

At 200 mg/ kg/ day, hematocrit, hemoglobin, serum albumin, and albumin/ globulin ratio were decreased ( p< 0.05) in females. Additionally, relative ( to body) liver weights were increased and slight diffuse lobular hepatocyte hypertrophy was observed in two males and two females at this dose compared to 0/ 0 in the controls. Absolute and relative ( to body) thymus weights were decreased. A greater incidence of hemosiderin pigment deposits were observed in the spleen of males and the lesion was less severe than at 1000 mg/ kg/ day.

The only effects observed at 40 mg/ kg were decreased ( p  0.05) hematocrit, hemoglobin, serum albumin, and relative ( to body) thymus weight in females ( p  0.05).

The LOAEL is 200 mg/ kg/ day based on decreases in hematocrit, hemoglobin, serum albumin and albumin/ globulin ratio in females and increases in liver to body weight ratios, hepatocyte hypertrophy ( both sexes) and hemosiderin pigment deposits in the spleen of males. The NOAEL is 40 mg/ kg/ day.

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

2.2. Chronic/ Carcinogenicity Studies 2.2.1. Murata, K. ( 2001) KIF 230 Technical: chronic toxicity study and oncogenicity study in rats. Biosafety Research Center, Shizuoka, Japan. Laboratory Experimental No.: 3822 ( 001 208), September 14, 2001. MRID 45835017. Unpublished. EXECUTIVE SUMMARY  In this combined chronic toxicity/ carcinogenicity study ( MRID 45835017), KIF 230 technical ( benthiavalicarb; 88.8 89.1% a. i.; dose levels adjusted for purity; Lot No.: G51 08 158) was administered in the diet for up to 2 years to Fischer F344/ DuCrj rats Page 42 of 87 ( 80/ sex/ dose) at nominal concentrations of 0, 50, 200, 5000, or 10,000 ppm ( equivalent to 0/ 0, 2.5/ 3.2, 9.9/ 12.5, 249.6/ 318.2, and 518.3/ 649.4 mg/ kg/ day in males/ females). Ten rats/ sex/ dose were sacrificed at Weeks 26, 52, and 78, and the remaining survivors were sacrificed at Week 104.

No treatment related effects were observed on mortality, clinical signs, food consumption, food efficiency, ophthalmoscopic examination, or on hematology parameters.

At   5000 ppm, nephrotoxicity and hepatotoxicity were observed. In the kidney, absolute and relative ( to body) weights were increased throughout the study in both sexes. At Week 52, increased incidences of basophilic tubules were noted in the males. At Week 104, increased incidences of the following lesions were observed in the kidney: ( i) chronic nephropathy and tubular dilatation in the males; ( ii) glomerulosclerosis, calculus, and hyaline droplets in both sexes; and ( iii) lymphocytic cellular infiltration, basophilic tubules, hyaline casts, and brown pigment deposits in the females. Urinary protein levels were increased at Week 52; however, this transient effect may have been incidental. Liver findings included increased gamma glutamyltranspeptidase in the males at Weeks 26 and 104 and in the females throughout the study. Increased levels of total and free cholesterol and phospholipids were observed in the females throughout the study. Absolute and relative ( to body) liver weights were increased throughout the study in both sexes. Grossly, hepatic enlargement was noted at Week 52 ( in males only) and at Week 78 ( in both sexes), and was also observed in the 10,000 ppm group at Weeks 26 and 52. Additionally, at Week 104, increased incidences were observed of brown and/ or white patches on the liver in the males, and red patches on the liver in the females. An increased incidence in hepatocellular hypertrophy was observed in both sexes at Weeks 26, 52, and 78; and in the females at Week 104 ( and in the 10,000 ppm males at Week 104). A fatty change was noted in the males at Week 52, in both sexes at Week 78, and in females at Week 104. Additionally at Week 104, increased incidences of foci/ area of cellular alteration in the liver and spongiosis hepatis were observed in the males.

At 10,000 ppm, the following additional effects were observed in the kidney and liver. In the kidney, an increased incidence of granular kidney was noted grossly in males at Week 104. Glomerusclerosis, hyaline casts, and hyaline droplets were observed in the males at Week 52. Additionally, at Week 104, increased incidences of transitional cell hyperplasia, fibrosis, and lymphocytic cellular infiltration were noted in males. In both sexes, an increase in the amount of smooth endoplasmic reticulum was noted in the liver at Weeks 26 and 52. Only minor decreases in body weight and body weight gain were observed in this group.

The LOAEL is 5000 ppm ( equivalent to 249.6/ 318.2 mg/ kg/ day in males/ females), based on nephrotoxicity and hepatotoxicity in both sexes. The NOAEL is 200 ppm ( equivalent to 9.9/ 12.5 mg/ kg/ day in males/ females).

There was a treatment related increase in tumor incidence when compared to controls. Increased incidences of hepatocellular adenoma were observed (% treated vs % controls) in the 10,000 ppm males at Weeks 78 ( 10 vs 0; NS) and 104 ( 14 vs 2; p  0.05), but fell within the range of historical controls ( 0 18%). Mutagenicity tests of this compound and concurrently submitted mechanistic studies ( MRID 45835021 and 45869715) suggest that KIF 230 is acting as a promoter, but not an initiator, in the development of rat liver neoplasia. Additionally, uterine adenocarcinoma was Page 43 of 87 increased at   5000 ppm at Weeks 78 ( 20% vs 0%; NS) and 104 ( 20 22% vs 6%; p  0.05) and exceeded the historical control range ( 0 8%). Increased ( p  0.05) estradiol levels were observed in the   5000 ppm females at Weeks 26 and 78 (  27 71%). However, the Sponsor indicated that a more excessive increase in estradiol levels with corroborating histological effects would have been expected if elevated hormone levels were the causative factor in the uterine neoplasia. The Sponsor stated that the uterine adenocarcinoma was most likely due to a direct effect on the organ. Dosing was considered to be adequate based on increased lesions in the kidneys and liver.

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.

2.2.2. Murata, K ( 2001) KIF 230 Technical: Oncogenicity study in mice. Biosafety Research Center, Foods, Drugs and Pesticides ( An Pyo Center) Arahama Shioshinden, Fukude Cho Iwata Gun, Shizuoka, Japan. Laboratory Project ID.: 3823 ( 001 209), September 7, 2001. MRID 45835016. Unpublished.

EXECUTIVE SUMMARY  In a carcinogenicity study ( MRID 45835016), benthiavalicarb ( KIF 230); 88.8 89.1% ( w/ w), dose levels adjusted for purity, Lot/ Batch # G51 08 158) was administered to 70 SLC: B6C3F1 ( C57BL/ 6xC3H)[ SPF] mice/ sex/ group in the diet at dose levels of 0, 20, 100, 2500, or 5000 ppm ( equivalent to 0/ 0, 2.7/ 3.7, 13.7/ 18.6, 358.4/ 459.3, 731.3/ 927.8 mg/ kg/ day [ M/ F], respectively) for 104 weeks, with interim sacrifices of 10 mice/ sex/ dose at 52 and 78 weeks. No adverse treatment related effects were noted on food consumption.

At   2500 ppm, greater incidences of clinical signs of toxicity ( abdominal masses and distention, and pallor) were observed in males during Weeks 79 104. Body weight, overall ( Week 0 104) body weight gain, and overall ( Weeks 0 52) food efficiency were also decreased ( p  0.05) in males. At 5000 ppm, survival at Week 104 was significantly ( p  0.01) decreased in males. At 5000 ppm, wasting, piloerection, and tachypnea were clinically observed in males during Weeks 79 104. At   2500 ppm, hepatotoxicity and thyroid gland toxicity were observed. Absolute and relative to body liver weights were increased ( p  0.05) in both sexes. The incidences of brown patches and nodules of the liver in both sexes, and enlarged liver in females were increased ( p  0.05) at   2500 ppm. At 5000 ppm, red patches in males and white patches in females were grossly observed in the liver. Slight intermediate fatty change, slight hepatocyte hypertrophy, slight to marked foci of cellular alteration were observed in both sexes. Necrosis, single cell necrosis, anisonucleosis, cellular infiltration, lymphocyte infiltration, multinucleated cells, bile duct proliferation, extramedullary hematopoiesis, and microgranuloma were observed in males at   2500 ppm and were slight in severity. Additionally, slight to moderate accumulation of macrophages were observed in males at   2500 ppm. At 5000 ppm, slight necrosis, single cell necrosis, anisonucleosis, and cellular infiltration were observed in females and slight angiectasis was observed in males.

In the thyroid gland, increased ( p  0.01) incidences of follicular cell hyperplasia in both sexes and dilated thyroid follicles in males at   2500 ppm and in females at 5000 ppm were observed. In the ovary, absolute and relative to body weight were decreased ( p  0.05) and ovarian atrophy was observed in females. Page 44 of 87 Platelet count was increased ( p  0.05) at   2500 ppm in both sexes. Clotting time parameters were not measured; therefore, the functional impact of increased platelet count was not demonstrated. However, the platelet count difference was corroborated by an increase in the number of megakaryocytes in the 5000 ppm males. The toxicological relevance of this finding was considered equivocal.

The only effects observed at 100 ppm were abdominal distention during Weeks 79 104 and slight microgranuloma in the liver in males.

The LOAEL is 2500 ppm ( equivalent to 358.4/ 459.3 mg/ kg/ day [ M/ F]) based on clinical signs ( abdominal masses and distention, and pallor) in males, decreased body weight gain and food efficiency in males, gross liver lesions ( brown patches and nodules in both sexes, and enlarged liver in females), increased absolute and relative ( to body) liver weight in both sexes, nonneoplastic liver lesions ( hepatocyte hypertrophy, hepatic intermediate fatty change, and hepatic foci of cellular alteration in both sexes, and anisonucleosis, necrosis, single cell necrosis, multinucleated cells, accumulation of macrophages, cellular infiltration, lymphocyte infiltration, bile duct proliferation, and extramedullary hematopoiesis in males), thyroid follicular cell hyperplasia in both sexes, and dilated thyroid follicles in males. The NOAEL is 100 ppm ( equivalent to 13.7/ 18.6 mg/ kg/ day in M/ F).