UNITED STATES ENVIRONMENTAL PROTECTION AGENCY WASHINGTON, D. C. 20460 OFFICE OF PREVENTION, PESTICIDES AND TOXIC SUBSTANCES November 30, 2005 MEMORANDUM SUBJECT: Bitertanol. Revised HED Chapter of the Tolerance Reassessment Eligibility Decision Document ( TRED). PC Code: 117801, Decision #: 343558, DP Barcode: D323894.

Regulatory Action: Tolerance Reassessment Eligibility Decision Risk Assessment Type: Single Chemical, No Aggregate FROM: Christina Jarvis, Risk Assessor Reregistration Branch 2 Health Effects Division ( 7509C)

AND Dave Anderson, Ph. D., Toxicologist Yvonne Barnes, Product Chemist Samuel Ary, Residue Chemist/ Dietary Exposure Reregistration Branch 2 Health Effects Division ( 7509C)

THROUGH: Alan Nielsen, Branch Senior Scientist Reregistration Branch 2 Health Effects Division ( 7509C)

TO: Rosanna Louie, Chemical Review Manager Special Review and Reregistration Division ( 7508C) ii Table of Contents 1.0 Executive Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 1 of 57 2.0 Ingredient Profile . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 3 of 57 2.1 Summary of Registered Uses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 4 of 57 2.2 Structure and Nomenclature . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 4 of 57 2.3 Physical and Chemical Properties . . . . . . . . . . . . . . . . . . . . . . . . . . Page 5 of 57 3.0 Metabolism Assessment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 6 of 57 3.1 Comparative Metabolic Profile . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 6 of 57 3.2 Nature of the Residue in Foods . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 6 of 57 3.2.1. Description of Primary Crop Metabolism . . . . . . . . . . . . . . Page 6 of 57 3.2.2 Description of Livestock Metabolism . . . . . . . . . . . . . . . . . Page 6 of 57 3.2.3 Description of Rotational Crop Metabolism . . . . . . . . . . . . Page 7 of 57 3.3 Environmental Degradation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 7 of 57 3.4 Tabular Summary of Metabolites and Degradates . . . . . . . . . . . . . . Page 7 of 57 3.5 Summary of Residues for Tolerance Expression and Risk Assessment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 7 of 57 3.5.1 Tabular Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 7 of 57 4.0 Hazard Characterization/ Assessment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 8 of 57 4.1 Hazard Characterization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 8 of 57 4.2 FQPA Hazard Considerations . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 17 of 57 4.2.1 Adequacy of the Toxicity Data Base . . . . . . . . . . . . . . . . . Page 17 of 57 4.2.2 Evidence of Neurotoxicity . . . . . . . . . . . . . . . . . . . . . . . . . Page 18 of 57 4.2.3 Developmental Toxicity Studies . . . . . . . . . . . . . . . . . . . . . Page 18 of 57 <IP_ADDRESS> Rat Developmental Toxicity with Bitertanol [ Long Evans rats] . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 18 of 57 <IP_ADDRESS> Rat Developmental Toxicity with Bitertanol [ Sprague Dawley rats] . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 20 of 57 <IP_ADDRESS> Rabbit Developmental Toxicity with Bitertanol [ Chinchilla rabbits] . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 21 of 57 <IP_ADDRESS> Rabbit Developmental Toxicity with bitertanol [ Himalayan rabbits] . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 22 of 57 <IP_ADDRESS> Rabbit Developmental Toxicity with bitertanol [ Himalayan rabbits] . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 23 of 57 4.2.4 Reproductive Toxicity Study Bitertanol . . . . . . . . . . . . . . Page 25 of 57 4.2.5 Additional Information from Literature Sources . . . . . . . . Page 27 of 57 4.2.6 Pre and/ or Postnatal Toxicity . . . . . . . . . . . . . . . . . . . . . . . Page 29 of 57 <IP_ADDRESS> Determination of Susceptibility . . . . . . . . . Page 29 of 57 <IP_ADDRESS> Degree of Concern Analysis and Residual Uncertainties for Pre and/ or Post natal Susceptibility . . . . . . Page 29 of 57 4.3 Recommendation for a Developmental Neurotoxicity Study . . . . . Page 30 of 57 4.3.1 Evidence that supports requiring a Developmental Neurotoxicity study iii . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 30 of 57 4.3.2 Evidence that supports not requiring a Developmental Neurotoxicity study . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 30 of 57 <IP_ADDRESS> Rationale for the UF DB . . . . . . . . . . . . . . . . Page 30 of 57 4.4 Hazard Identification and Toxicity Endpoint Selection . . . . . . . . . Page 30 of 57 4.4.1 Acute Reference Dose ( aRfD)  Females age 13 49 . . . . . . Page 30 of 57 4.4.2 Acute Reference Dose ( aRfD)  General Population . . . . . Page 31 of 57 4.4.3 Chronic Reference Dose ( cRfD) . . . . . . . . . . . . . . . . . . . . Page 31 of 57 4.4.4 Incidental Oral Exposure ( Short Term, 1 30 days), and Intermediate Term, 1 month 6 months) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 32 of 57 4.4.5 Dermal Absorption . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 32 of 57 4.4.6 Dermal Exposure ( Short, Intermediate and Long Term) . . Page 33 of 57 4.4.7 Inhalation Exposure ( Short, Intermediate and Long Term) Page 33 of 57 4.4.8 Margins of Exposure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 33 of 57 4.4.9 Recommendation for Aggregate Exposure Risk Assessments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 33 of 57 4.4.10 Classification of Carcinogenic Potential . . . . . . . . . . . . . . . Page 33 of 57 <IP_ADDRESS> Combined Chronic Toxicity/ Carcinogenicity Study in Rats . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 33 of 57 <IP_ADDRESS> Carcinogenicity Study in Mice . . . . . . . . . . . . . . . Page 35 of 57 4.5 Special FQPA Safety Factor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 37 of 57 4.6 Endocrine disruption . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 37 of 57 5.0 Public Health Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 38 of 57 5.1 Incident Reports . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 38 of 57 6.0 Exposure Characterization/ Assessment . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 38 of 57 6.1 Dietary Exposure/ Risk Pathway . . . . . . . . . . . . . . . . . . . . . . . . . . Page 38 of 57 6.1.1 Residue Profile . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 38 of 57 6.1.2 Acute and Chronic Dietary Exposure and Risk . . . . . . . . . Page 40 of 57 6.2 Water Exposure/ Risk Pathway . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 41 of 57 6.3 Residential ( Non Occupational) Exposure/ Risk Pathway . . . . . . . Page 41 of 57 7.0 Aggregate Risk Assessments and Risk Characterization . . . . . . . . . . . Page 41 of 57 8.0 Cumulative Risk Characterization/ Assessment . . . . . . . . . . . . . . . . . . Page 41 of 57 9.0 Occupational Exposure/ Risk Pathway . . . . . . . . . . . . . . . . . . . . . . . . . Page 42 of 57 10.0 Data Needs and Label Requirements . . . . . . . . . . . . . . . . . . . . . . . . . . Page 42 of 57 10.1 Toxicology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 42 of 57 10.2 Residue Chemistry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 42 of 57 APPENDICES A C . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 48 of 57 iv Page 1 of 58 1.0 Executive Summary Bitertanol [ beta([ 1,1' biphenyl] 4 yloxy) alpha( 1,1 dimethylethyl) 1H 1,2,4 triazole 1 ethanol] is a triazole fungicide used to control black sigatoka on banana and plantain plants. Bayer CropScience is the sole registrant of bitertanol. Bitertanol is marketed under the trade name Baycor ® and is formulated as an emulsifiable concentrate ( EC) and a suspension concentrate ( SC). The percentage of active ingredient in the end use formulations ranges from 30 percent ( EC) to 50 percent ( SC). Applications of bitertanol may be made aerially, by backpack sprayer, and by other types of equipment. The maximum single application rate is 0.13 pounds active ingredient per acre ( lbs ai/ A), and the maximum seasonal application rate is 1.06 lbs ai/ A.

There are no existing registrations for the use of bitertanol in the United States; therefore, this assessment represents the tolerance reassessment decision for bitertanol use on imported bananas only. Major countries that intend to export bananas or plantains that may contain bitertanol residues include Ecuador, Guatemala, Costa Rica, Columbia, and Honduras; minor countries include Mexico, Nicaragua, Peru, Venezuela, Dominican Republic, and Panama. The toxicological database for bitertanol is adequate when considered as a whole; however, when reviewed individually, many of the studies are marginally adequate to inadequate, with some studies dating back to the mid 1970s. Most of the studies were conducted prior to the publications of GLPs, and have other deficiencies. However, the Health Effects Division's ( HED's) Risk Assessment Review Committee ( RARC) has determined that the toxicological database for bitertanol is adequate for risk assessment purposes.

Bitertanol has low acute toxicity via the oral and dermal routes of exposure ( toxicity categories IV and III, respectively). An acceptable acute inhalation study is not available; however, since bitertanol is an import tolerance, none is needed for registration in the United States as an import tolerance. Bitertanol is a mild eye irritant ( toxicity category IV). It is neither a dermal irritant nor a dermal sensitizer.

Multi dose studies necessary for reregistration, when reviewed in combination with other studies, show sufficient consistency to classify studies as acceptable, although the studies are inadequate when reviewed in isolation. For example, a combination of two rat developmental studies and three rabbit developmental studies are acceptable for developmental toxicity in the rat and rabbit. Doses used and effects seen in a three generation reproduction study are similar to an acceptable 90 day study in rats; therefore, when combined with the 90 day study in rats, the three generation reproduction study is considered acceptable. Carcinogenicity studies in the rat and mouse are acceptable, and show no treatment related carcinogenicity at marginally toxic dose levels. Bitertanol is not mutagenic.

The toxicological database is adequate for FQPA hazard assessment. In submitted developmental toxicity studies, qualitative susceptibility was seen in the rat and rabbit at the same dose levels as maternal toxicity. There is no evidence of neurotoxicity among the guideline studies in the toxicity database; however, literature studies with bitertanol show limited neurotoxic effects. There was no indication of increased susceptibility in the rat reproduction study.

An acute reference dose ( aRfD) for the general population has not been selected for bitertanol. Page 2 of 58 An appropriate endpoint attributable to a single dose was not available in the toxicity database, including the developmental toxicity studies. The aRfD of 0.05 mg/ kg/ day for females 13 49 years of age is based on a No Observed Adverse Effect Level ( NOAEL) of 50 mg/ kg/ day and an uncertainty factor of 1000X ( 10X for interspecies extrapolation, 10X for intraspecies variation, and 10X database uncertainty factor due to lack of acute and subchronic neurotoxicity studies). There is no residual uncertainty with respect to pre and post natal toxicity based on the submitted developmental and reproduction studies. Thus, the special FQPA safety factor has been reduced to 1X. The aRfD is equivalent to the acute population adjusted dose ( aPAD).

The chronic RfD ( cRfD) of 0.00211 mg/ kg/ day is based on a NOAEL of 2.11 mg/ kg/ day and the combined 1000X uncertainty factor. Since the special FQPA safety factor has been reduced to 1X, the cRfD is equivalent to the chronic population adjusted dose ( cPAD).

No endpoints were selected for short, intermediate, and long term incidental oral, dermal, or inhalation exposure; therefore, these risk assessments were not conducted. Incidental oral and inhalation exposure endpoints are not necessary as there are no such exposures in the United States. A dermal exposure assessment is not necessary for an import tolerance. A cancer assessment is not required, as no dose related tumors were seen in the rat or mouse. An aggregate risk assessment is not required, as there is no exposure to residues of bitertanol through drinking water or residential uses.

A rat metabolism study indicates that bitertanol, at single low and high doses, is rapidly absorbed, excreted, and extensively metabolized. Both parent compound and numerous metabolites were detected. No banana metabolism studies are available; however, acceptable plant metabolism studies in apples and peanuts are available, as well as recently submitted metabolism studies in apples, cotton, and tomatoes. HED has determined that the nature of residues in bananas is likely similar to that in apples, peanuts, cotton, and tomatoes. Residues identified in these studies consist primarily of the parent compound, with traces of the metabolites bitertanol ketone and 4 hydroxybiphenyl. For risk assessment purposes, HED has determined that the bitertanol residues of concern in/ on bananas for tolerance expression and for risk assessment purposes are bitertanol ( parent compound only).

A single tolerance is established at 0.20 ppm for residues of bitertanol ( parent only) in/ on bananas ( whole), which also includes residues on plantains ( 40 CFR § 180.457). The Codex Alimentarius Commission ( Codex) has established a maximum residue limit for bitertanol per se in/ on bananas at 0.50 ppm. It is the Agency's policy to harmonize its tolerances with the levels established by the Codex, provided that the Agency has sufficient information to make a determination that the Codex maximum residue limits will be protective of the health of the U. S. public, and meet FFDCA standards. In addition, the available field trial studies suggest that a tolerance increase is advisable. Therefore, the established tolerance for bitertanol should be increased to 0.50 ppm to harmonize with Codex.

Since there are no existing U. S. registrations for bitertanol, the only exposure pathway considered in this risk assessment is for acute and chronic dietary exposure to bitertanol residues resulting from the consumption of whole bananas. Conservative acute and chronic dietary risk estimates were calculated using DEEM FCID   , incorporating the proposed tolerance increase of 0.50 ppm Page 3 of 58 ( mg/ kg/ day), the DEEM Version 7.81 default processing factor of 3.9x for dried bananas, and an assumption of 100% crop treated. For the supported use of bitertanol on imported bananas, acute and chronic dietary risk estimates do not exceed HED's level of concern ( i. e., risks are less than 100% of the acute/ chronic population adjusted dose) for any population subgroup.

Data gaps: Storage stability data for bitertanol in/ on bananas have not been submitted by the registrant; therefore, storage stability data are required to determine the stability of residues in bananas during cold storage. Field trial data for the suspension concentrate ( SC) formulation are required ( these field trials are currently being conducted by the registrant and are planned for completion in 2006). Additional data ( the number of trials, location of the trials within the countries, and the formulation type used) are required in order for the submitted field trials conducted in Costa Rica and Honduras to be acceptable.

With regards to toxicology data needs, an acute neurotoxicity battery and a 90 day neurotoxicity study with bitertanol are recommended. The requirement for a developmental neurotoxicity ( DNT) study is held in reserve, pending the submission and review of the acute and subchronic neurotoxicity studies.

2.0 Ingredient Profile P. Yvonne Barnes. Bitertanol: Summary of Product Chemistry Data for Tolerance Reassessment Eligibility Decision ( TRED) Document. October 25, 2005. DP Barcode D302878.

Bitertanol is a broad spectrum triazole fungicide used to control black sigatoka on banana and plantain plants. Bayer CropScience is the sole registrant of bitertanol. Bitertanol is marketed under the trade name Baycor ® and is formulated as an emulsifiable concentrate ( EC) and a suspension concentrate ( SC). The percentage of active ingredient in the end use formulations ranges from 30 percent ( EC) to 50 percent ( SC). Applications of bitertanol may be made aerially, by backpack sprayer, and by other sprayer equipment. The maximum single application rate is 0.13 pounds active ingredient per acre ( lbs ai/ A), and the maximum seasonal application rate is 1.06 lbs ai/ A. Application is foliar during fruit development. The minimum interval between treatments is seven days. Bitertanol is intended for use on bagged bananas only; however, bananas are not always bagged and plantains are rarely bagged.

There are no existing registrations for the use of bitertanol in the United States; therefore, this assessment represents the tolerance reassessment decision for bitertanol use on imported bananas only. Major countries that intend to export bananas or plantains that may contain bitertanol residues include Ecuador, Guatemala, Costa Rica, Columbia, and Honduras; minor countries include Mexico, Nicaragua, Peru, Venezuela, Dominican Republic, and Panama. Page 4 of 58 2.1 Summary of Registered Uses Table 2.1: Summary of Directions for Use of Bitertanol.

App. Timing, Type, and Equip. Formulation App. Rate ( lb ai/ A) Max. No. App. per Season Max. Seasonal App. Rate ( lb ai/ A) PHI ( days) Use Directions and Limitations Banana Apply as needed based on speed of leaf emergence during fruit development. Minimum retreatment interval of 7 days. Apply specified dosage as a foliar spray by air, tractor driven equipment, or knapsack sprayer in a minimum of 2 gallons of water per acre. Baycor ® 30% EC 0.13 lb ai/ A 8 1.06 lb ai/ A None

Baycor ® 50% SC 0.13 lb ai/ A 8 1.06 lb ai/ A None

2.2 Structure and Nomenclature Table 2.2: Bitertanol Structure and Nomenclature Chemical Structure Empirical Formula C20H23N3O2 Common name Bitertanol Trade name Baycor ®

IUPAC name 1( biphenyl 4 yloxy) 3,3 dimethyl 1( 1H 1,2,4 triazol 1 yl) butan 2 ol CAS name [ beta([ 1,1' biphenyl] 4 yloxy) alpha( 1,1 dimethylethyl) 1H 1,2,4 triazole 1 ethanol]

CAS Registry Number 55179 31 2 Chemical Class Azole Page 5 of 58 2.3 Physical and Chemical Properties Table 2.3: Physicochemical Properties Parameter Value Reference Molecular Weight 337.42 grams/ mole MRID 46655901 Melting point/ range 145 155 ° C MRID 46655901 Density 1.16 g/ mL at 20 ° C MRID 46655901 Water solubility 1.1 mg/ L at 20 ° C ( isomer A) MRID 46655901 2.7 mg/ L at 20 ° C ( isomer B)

Solvent solubility ( temperature not specified) Propan 2 ol 1 5g/ 100g, toluene 1 5g/ 100g, cyclohexane 5 10g/ 100g, methylene chloride 10 20g/ 100g, Stable in aqueous acid and alkaline solutions. Merck 12th Edition Vapor pressure 0.00001 mBar at 20 ° C Merck 12th Edition Octanol/ water partition coefficient, log P K ow Kow logP = 4.04 at 20 ° C ( isomer A) MRID 46655901 Kow logP = 4.1 at 20 ° C ( isomer B)

UV/ visible absorption spectrum Peak maxima at 255 nm MRID 46655901 Page 6 of 58 3.0 Metabolism Assessment 3.1 Comparative Metabolic Profile Based upon the metabolites identified, the investigators concluded that BAYCOR was metabolized in the rat via mono ring hydroxylation ( p hydroxy BAYCOR I and p hydroxy BAYCOR II), di ring hydroxylation ( m, m dihydroxy BAYCOR I and m, m dihydroxy BAYCOR I), aryl O methylation and aliphatic hydroxylation ( p hydroxy m methoxy BAYCOR, BAYCOR alcohol, p hydroxy BAYCOR alcohol I and p hydroxy BAYCOR alcohol II), ether cleavage and aliphatic oxidation to carboxylic acids. The investigators noted the absence of metabolites ( e. g., precursors to identified metabolites) typically expected to occur but hypothesized that these would likely be short lived intermediates thereby precluding accumulation and detection. A metabolism pathway was proposed based upon the study findings ( Appendix C).

The rat metabolism study indicates that bitertanol, at single low and high doses, is rapidly absorbed, excreted and extensively metabolized. Recovery of radioactivity was acceptable ( 92 104%). The studies affirmed the relatively rapid absorption of [ 14C phenyl UL] BAYCOR ( Bitertanol) and that excretion was primarily via the feces ( 81 98% and 92%, respectively) over the 168 hr experimental period. Urinary excretion ( 4 11%) was secondary to fecal elimination. Saturated absorption at the high dose ( 1000 mg/ kg) was evident by a plateau in plasma radioactivity levels ( and by estimated kinetic parameters) and an increase in the fecal excretion of parent compound. A minor gender related variance in absorption and elimination constants and plasma elimination halftime was observed for the multiple dose group but not for other treatment groups. No other gender related variability was observed. The study reported no excretion via expired air. Tissue radioactivity was minimal and not suggestive of bioaccumulation of the test material or its metabolites.

Since the triazole ring was not labeled, the triazole metabolite, if present, was not detected. [ Triazole could not be detected in other triazole ring containing pesticides when the triazole ring was not labeled.]

3.2 Nature of the Residue in Foods 3.2.1. Description of Primary Crop Metabolism The nature of the residue in plants is adequately understood based on acceptable metabolism studies on apples and peanuts. The residues consisted primarily of the parent compound with traces of the metabolites bitertanol ketone and 4 hydroxybiphenyl. The Agency has concluded that the residue of concern is the parent compound ( S. Ary, D302882, 11/ 7/ 2005).

3.2.2 Description of Livestock Metabolism Bitertanol is not intended for application to livestock exported to the U. S. In addition, there are no feedstuffs associated with bananas. Therefore, no residue chemistry data are required under this guideline topic. Page 7 of 58 O N N N O O H 3.2.3 Description of Rotational Crop Metabolism Rotational crop studies are not required for uses of pesticides on bananas ( HED SOP 96.6).

3.3 Environmental Degradation Bitertanol is registered for use on imported bananas only; therefore, environmental fate data are not required..

3.4 Tabular Summary of Metabolites and Degradates Table 3.4: Chemical Names and Structures of Metabolites of Bitertanol.

Common Name Chemical Name Structure bitertanol ketone 1[( 1,1 biphenyl) 4 yloxy] 3,3 dimethyl 1( 1H 1,2,4 triazole 1 yl) 2 butanone p hydroxybiphenyl 4 hydroxybiphenyl 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 Plants Primary Crop bitertanol bitertanol Rotational Crop n/ a n/ a Livestock Ruminant n/ a n/ a Poultry n/ a n/ a Drinking Water n/ a n/ a Page 8 of 58 4.0 Hazard Characterization/ Assessment 4.1 Hazard Characterization Bitertanol is a triazole fungicide used for imported bananas only, that was registered about 1994 1996. Bitertanol is among a class of over 25 fungicides. The literature noted that some triazole pesticides caused behavioral effects. Two analogs of bitertanol, triadimefon and triadimenol caused effects on operant behavior. An effect on operant behavior caused by acute dosing with bitertanol differed from that resulting from triadimefon and triadimenol exposure. A study in mice suggested hyperactivity effects while other studies showing effects on operant behavior, suggesting an effect resembling hyperactivity, but no effects on motor activity [ Allen and MacPhail, 1993, Crofton, 1996 and FAO and WHO Working Groups, 1998]. However, the guideline studies on bitertanol showed no evidence of neurotoxic effects.

The acute oral LD50 study with bitertanol show a toxicity category IV for acute toxicity, but toxicity category III for acute dermal, largely because of the low highest dose tested. The acute inhalation study is unacceptable because the particle size in the exposure aerosol was not described. Eye and skin irritant were toxicity category IV and skin sensitization studies were negative.

Pre natal studies showed no increased offspring quantitative susceptibility, but show qualitative susceptibility in the form of malformations and post implantation loss. The reproduction study showed no increased qualitative or quantitative susceptibility. In the development toxicity studies, although none of the malformations were statistically significant in the rats or rabbits, in combination with the increased resorptions and nominally increased malformations in rabbits and nominally increased malformations in rats, bitertanol is classed as a teratogen at maternally toxic dose levels. The developmental toxicity studies in the rat show developmental effects in the form of stunted fetuses and nominally increased malformations, such as kinked tail, cleft palate, rib malformations and delayed ossification at maternally toxic dose levels in the Long Evans rat, but the Sprague Dawley rat developmental study conducted at lower doses showed only increased 14th rib and no malformations. The NOAEL in the Long Evans and Sprague Dawley rat for developmental and maternal toxicity were identical. In the Chinchilla rabbit, the developmental effects in the from of post implantation loss, decreased fetal weights and rib anomalies occurred at the maternal toxic dose levels. However, in two Himalayan rabbits studies, decreased fetal weight, increased resorptions, and cleft palate, pigeon chests, epignathus, aplasia of several lung lobes and aplasia of the left lung and hypoplasia of the right lung were noted. The postimplantation loss at the LOAEL is considered a qualitative susceptibility for fetuses [ more severe than maternal weight decrement]. The study on reproduction showed pup weight decrements and parental weight decrements at the same dose levels.

Subchronic studies in rats showed body weight decreases and elevations in clinical chemistry parameters and liver effects. Subchronic studies in dogs showed prostate weight decrement, and skin and mucus membrane irritation at the LOAEL and at the HDT body weight and thymus weight decrement. The multi exposure inhalation studies were all unacceptable due to inadequate explanation of the inhalation particle size and other inhalation conditions. However, regardless of the particle size, none to these inhalation studies in the rat or dog showed the mucosal irritation Page 9 of 58 seen in the 90 day dog study.

The subchronic studies showed effects at slightly lower dose levels than the maternal toxic dose levels in the developmental studies, but the NOAEL was comparable with that seen in the chronic studies. Effects on the body weight and liver were common effects seen in the rat, mouse and dog. However, the dog subchronic study was the only study showing dose related prostate weight decrement. The prostate weight decrement and severe mucosal irritation were not supported by the chronic dog study.

Chronic studies in the rat, mouse and dog with bitertanol showed body weight decrement and food efficiency decrement at the LOAEL and at higher dose levels liver pathology, stomach hyperplasia, and some indication of unconfirmed kidney effects. In some of the studies, liver enzymes and cholesterol levels were increased at doses resulting in liver pathology. Due to dose selection the subchronic study in dogs showed a slightly lower NOAEL than the chronic study in dogs. The 21 day dermal rabbit study showed no systemic effects at the highest dose tested and only minor skin irritation.

Bitertanol shows no dose related or treatment related increased incidence in carcinogenicity in the rat or mouse carcinogenicity studies. Both studies were classified as acceptable. However, both studies failed to analyze the dietary concentration of the test material. Marginal toxicity at the highest dose level shown by body weight decrement and clinical chemistry results indicated that the animals were dosed and the effects were supported by the 90 day studies in rats. The highest dose levels used were marginally adequate and all required tissues were not evaluated among other deficiencies that did not negate the studies. Both studies were conducted prior to GLP publication.

A battery of acceptable mutagenicity studies with bitertanol were universally negative for genetic damage and mutations. The dominant lethal study in mice was marginally acceptable and negative. Only one non toxic dose level was used [ 1000 mg/ kg, 1/ 4 of the mouse LD50]. The remaining mutagenicity studies meet the criteria for pre 1996 guideline studies.

HED recommends that an acute neurotoxicity battery, and a 90 day neurotoxicity study with bitertanol be submitted. The requirement for a developmental neurotoxicity toxicity study is held in reserve, pending the submission and review of the acute and subchronic neurotoxicity studies. The 10X FQPA factor is reduced to 1X and a 10X database uncertainty factor is applied to acute and chronic dietary endpoints to account for the lack of acute and subchronic neurotoxicity studies.

Bitertanol is rapidly absorbed and excreted in the feces and urine of the rat. Only about 6% 11% of the absorbed dose is excreted in the urine while most of the remaining dose is excreted through bile into the feces. Analysis of the feces showed 14 metabolites and conjugates including metabolites of the two enantiomers of bitertanol [ Appendix C]. The metabolic profile did not differ between the urine and feces except small amounts of parent was excreted in feces but not in the urine. About 50% of the dose remained unextracted in feces after attempts with multiple extractants. No radioactivity was noted in the breath of rats. There was evidence of metabolic saturation at the 1000 mg/ kg dose, but not of accumulation. Page 10 of 58 Metabolism studies with bitertanol [ BAYCOR] showed extensive metabolism. Based upon the metabolites identified, the investigators concluded that BAYCOR was metabolized via mono ring hydroxylation ( p hydroxy BAYCOR I and p hydroxy BAYCOR II), di ring hydroxylation ( m, mdihydroxy BAYCOR I and m, m dihydroxy BAYCOR I), aryl O methylation and aliphatic hydroxylation ( p hydroxy m methoxy BAYCOR, BAYCOR alcohol, p hydroxy BAYCOR alcohol I and p hydroxy BAYCOR alcohol II), ether cleavage and aliphatic oxidation to carboxylic acids. The investigators noted the absence of metabolites ( e. g., precursors to identified metabolites) typically expected to occur but hypothesized that these would likely be short lived intermediates thereby precluding accumulation and detection. A metabolism pathway was proposed based upon the study findings ( Appendix C, Figure 1).

Table 4.1a: Acute Toxicity Data on Bitertanol ( PC 117801)

Guideline No./ Study type MRID No. Results Toxicity category 870.1100 Acute oral toxicity/ rats ( 95.6%) 00025641, 00025640 [ 1977] [ # 51413] LD50 > 5000 mg/ kg IV 870.1200 Acute dermal toxicity/ rabbits ( 96.5%) 00025641 [ 1977] [# 51413] LD50 > 2000 mg/ kg III 870.1300 Acute inhalation toxicity/ rats ( 97%) 79/ ISK034/ 387 00025641 [ 1977], 00025653 [ 1979] [# 51413] For MRID# 0025641; Study unacceptable; undefined particle size & doses administered not clear. For MRID# 00025653; Study unacceptable; undefined particle size. LC50 > 5.16 mg/ L No indication of mucosal membrane irritation.

870.2400 Acute eye irritation/ rabbit ( 93.3%) 00025641,0 0025640 [ 1977] [# 51413] Mild irritation IV 870.2500 Acute dermal irritation/ rabbit ( 93.3%) 79ILK8/ 056 00025641,0 0025640 [ 1977] [# 51413] Non irritating IV 870.2600 Skin sensitization/ GP ( 96.5%) 00114277 [ 1981] [# 80324] Not a dermal sensitizer in the Magnsson and Kligman Test  Page 11 of 58 Table 4.1b: Subchronic, Chronic, Developmental, Reproductive and other Toxicity Profile on Bitertanol Guideline No./ Study Type MRID No. ( year)/ Classification / Doses Results 870.3100 90 Day oral toxicity ( SD rat) Acceptable 00157853 ( 1981) TXR# 0005568 0, 40, 200, 1000 ppm M/ F 0/ 0, 3.1/ 3.6, 15.9/ 18.6, 81.8/ 88.2 mg/ kg/ day.

No opthalmic examination NOAEL = 40 ppm ( M/ F: 3.1/ 3.6 mg/ kg/ day) LOAEL = 200 ppm ( M/ F: 15.9/ 18.6 mg/ kg/ day) based on decreased body weight and body weight gain in males and females. At 1000 ppm ( M/ F: 81.8/ 88.2 mg/ kg/ day) slightly decreased hematological parameters and slightly elevated LDH, SGOT, ALP ( 20%  50%) and increased cholesterol ( 117%). Increased incidence of hyperkeratotic epithelia of the esophagus and glandular stomach and increased liver and adrenal weights and liver histopathology.

870.3100 90 Day oral toxicity ( Wistar rat) Unacceptable ( Contractor) No need to repeat study. 00025647 ( 1976) TXR# 006018, p 13 Doses: 0, 30, 100, 300 ppm; M/ F: 0/ 0, 2.46/ 3.25, 8.15/ 10.15, 24.8/ 31.6 mg/ kg/ day N= 100 ppm ( M/ F: 8.2/ 10.2 mg/ kg/ day) L= 300 ppm ( M/ F: 24.8/ 31.6 mg/ kg/ day) based on marginal body weight decrease in males and females ( 6 8%). Decreased body weight gain in males and females ( 8 12%) by week 13.

Non guideline Two week oral toxicity of liver ( rats) 00163455 ( 1985) Acceptable/ Nonguideline: 0, 30, 100, 300 mg/ kg/ day Body weight decrement was seen 30 mg/ kg/ day in females and at 100 mg/ kg/ day in males. Microsomal enzymes [ Ndemethylase and cytoplasic granules were seen at 100 mg/ kg/ day and higher in females. Councilman bodies were seen in liver from females at 100 mg/ kg/ day and in males at 300 mg/ kg/ day. Bile duct proliferation was seen at 100 mg/ kg/ day and higher doses and isolated mitoses was in male and female livers at 30 mg/ kg/ day.

870.3200 21/ 28 Day dermal toxicity ( NZ rabbit) Acceptable 00149829 ( 1984) TXR# 004638 Supp. 11 Lner' 88 0, 50, 250 mg/ kg/ day NOAEL= 250 mg/ kg/ day for systemic effects. LOAEL= None, but slight skin irritation was seen at 250 mg/ kg/ day.

870.3150 90 Day oral toxicity ( dog) Acceptable 00025648 ( 1979) TXR# 006018 0, 1, 5, 25 mg/ kg/ day NOAEL= 1 mg/ kg/ day LOAEL= 5 mg/ kg/ day based on dose related decreased prostate weight, and dermal, conjunctival and mucosal irritation effects in males and females.

870.3150 21 Day Inhalation toxicity ( rats) 00025650 ( 1977) TXR# 006018; Unacceptable/ upgradab le on submission of acceptable particle size and other data on inhalation characteristics. Stated exposure levels [ unverified]: 0, 0.0179, 0.0693, 0.1979 mg/ L At 0.198 mg/ L, male body weight [ 94%] and body weight gain [ 69%] were statistically significantly less than control. Male heart weights were significantly decreased at 0.198 mg/ L No other possible treatment related effects were supported by histopathology in males or females, including irritation of the mucosal membranes. The study authors claimed a NOAEL/ LOAEL of 0.0693/ 0.198 mg/ L. Table 4.1b: Subchronic, Chronic, Developmental, Reproductive and other Toxicity Profile on Bitertanol Guideline No./ Study Type MRID No. ( year)/ Classification / Doses Results Page 12 of 58 ~ 870.3465 21 Day Inhalation toxicity ( dogs) 00025643 ( 1977) Unacceptable/ upgradab le on submission of acceptable particle size and other data on inhalation characteristics. Doses administered: 0.0288 mg/ L for 21 days and after 10 days exposed to 0.0471 mg/ L for 21 days in a whole body chamber. The dogs showed no effects from either exposures, including irritation of the mucosal membranes noted in the 90 day oral study in dogs. However, this study is unacceptable because it was inadequately described and data no particle sizes or concentration of the test material in the breathing zone of the dogs were submitted ~ 870.4100a Chronic Toxicity ( rats) See 870.4300 870.4100b Chronic toxicity ( Dogs) 00157465, 00157466 ( 1983), 401632 01 ( 1986), 02 ( 1986), 03 ( 1987) & 40186401 ( 1987) Studies combined Acceptable 0, 10, 40 160 ppm ( 0, 2.11, 8.18, 32.24 mg/ kg/ day) for 2 years and 0, 3, 25 ppm ( 0, 0.78, 7.14 mg/ kg/ day for 1 year and 200 ppm ( 50.93 mg/ kg/ day) for 86 weeks. NOAEL = 10 ppm ( 2.11 mg/ kg/ day) LOAEL = 40 ppm ( 8.18 mg/ kg/ day) based on adrenal vacuolation in the 2 year study, supported by adrenal vacuolation, and 160 and 200 ppm eye and liver effects.

~ 870.4200a Carcinogenicity ( rats) See 870.4300 ~ 870.4200b Carcinogenicity ( Mice) 00114280 ( 1978) TXR# 0002588 0, 20, 100, 500 ppm ( M: 0, 5, 25, 129 mg/ kg/ day; F: 0, 7, 37, 183 mg/ kg/ day) NOAEL = 100 ppm [ M/ F: 25/ 37 mg/ kg/ day] LOAEL = 500 ppm [ M/ F: 129/ 183 mg/ kg/ day] based on marginal body weight decrement in males and females through out the study and other statistically significant, but marginal effects, such as eosinophilic foci in livers of females. Increased liver wt & GPT in females and increased ALP & urea in males.

No dose related carcinogenicity was seen. Table 4.1b: Subchronic, Chronic, Developmental, Reproductive and other Toxicity Profile on Bitertanol Guideline No./ Study Type MRID No. ( year)/ Classification / Doses Results Page 13 of 58 ~ 870.4300 Combined chronic/ carcinogeni city ( rat) 00114279 ( 1981) TXR# 0002588 0, 20, 100, 500 ppm [ M: 0, 1.01, 4.89, 25.54 mg/ kg/ day: F: 0, 1.32, 6.55, 33.53] mg/ kg/ day] NOAEL = 100 ppm [ M/ F: 4.89/ 6.55 mg/ kg/ day] LOAEL = 500 ppm [ M/ F: 25.54/ 33.53 mg/ kg/ day] based on nominal decreased body weight ( 8%), body weight gain ( M/ F 11/ 13%) and nominal food efficiency in males and females ( 9%) throughout the treatment period. Statistically significant hematological effects, but within normal ranges were seen and clinical chemistry values were statistically significantly changed, but within normal ranges, such as decreased urea, increased creatinine.

No dose related tumors were seen in the study.

~ Two rat developmental toxicity studies in Long Evans and Sprague Dawley rats below 00154936 ( 1977) & 00114283 ( 1981) together are acceptable Both studies together are acceptable. Analysis of doses were conducted in neither study, among other deficiencies. However, since both studies showed the similar NOAEL/ LOAEL, it can be concluded that the dose levels were reasonably accurate, and other deficiencies are not sufficient to show unacceptability. Malformations were seen at the HDT in the Long Evans rat.

~ 870.3700a Prenatal developmental in ( Long Evans rats) 00154936 ( 1977) Acceptable combined with 00114283 ( 1981) 0, 10, 30, 100 mg/ kg/ day No dose concentration analysis Maternal NOAEL= 10 mg/ kg/ day LOAEL = 30 mg/ kg/ day based on dose related maternal weight gain decrement. Devel NOAEL = 10 mg/ kg/ day LOAEL = 30 mg/ kg/ day based on stunted fetuses and at 100 mg/ kg/ day malformations, kinked tail, cleft palate, rib malformations and delayed ossification.

~ 870.3700a Prenatal Development in CR Sprague Dawley Japan rats Report# ; 80571 00114283 ( 1981) Acceptable combined with 00154936 ( 1977) 0, 10, 25, 65 mg/ kg/ day TXR# 002588 Maternal NOAEL= 10 mg/ kg/ day LOAEL= 25 mg/ kg/ day based on statistically significant decreased body weight gain decrement during dosing period ( GD 6 15).

Devel NOAEL= 10 mg/ kg/ day. LOAEL= 25 mg/ kg/ day increased 14th rib 32% and 70% at 65 mg/ kg/ day.

Three developmental toxicity studies in Chinchilla and Himalayan rabbits below 40490801, 40829401 ( 1987), 00149830 & 40163205 [ supp] ( 1983) & 00154937 SOP ( 1982) and 00114284 ( 1982) together with the supplementary information are acceptable for a guideline developmental toxicity in the rabbit. Nominal increases in malformations were seen at 100 and 150 mg/ kg/ day. Table 4.1b: Subchronic, Chronic, Developmental, Reproductive and other Toxicity Profile on Bitertanol Guideline No./ Study Type MRID No. ( year)/ Classification / Doses Results Page 14 of 58 ~ 870.3700b Prenatal developmental in ( Chinchilla rabbits) Report# 065283: 94704 40490801, 40829401 ( 1987) Acceptable 0, 10, 50, 250, 0 ,150 mg/ kg/ day Maternal NOAEL = 50 mg/ kg/ day LOAEL = 150 mg/ kg/ day based on decreased body weight gain and reduced food consumption. At 250mg/ kg/ day only one dam showed 8 normal fetuses, 4/ 16 dams died, 11/ 16 of the remaining dams showed complete resorption.

Devel. NOAEL = 50 mg/ kg/ day LOAEL = 150 mg/ kg/ day based increased post implantation loss, decreased fetal weight, skeletal anomalies ( abnormally ossified ribs and sternebrae and fused ribs).

~ 870.3700b Prenatal Developmental in Himalayan rabbits Report# 10979; 82239 Acceptable 00114284 ( 1982) Acceptable 0, 10, 30 ,100 mg/ kg/ day TXR# 002588 Maternal NOAEL= 30 mg/ kg/ day LOAEL= 100 mg/ kg/ day based on decreased maternal weight ( 85.5g versus 1.0 g) GD 6 18 Devel NOAEL= 30 mg/ kg/ day LOAEL= 100 mg/ kg/ day based on decreased fetal weight and increased resorptions and 3 litters with nominally increased uncommon malformations ( epignathus, aplasia of several lung lobes & aplasia left lung and hypoplasia of right lung verus none in controls).

~ 870.3700b Prenatal Developmental in Himalayan rabbits Report# Br11548; 86432 00149830 & 40163205 [ supp] ( 1983) & 00154937 SOP ( 1982) Unacceptable 0, 10, 30,100 mg/ kg/ day TXR# 004683 Maternal NOAEL= 30 mg/ kg/ day LOAEL= 100 mg/ kg/ day based on marginal decreased maternal body weight GD 6 18 ( 60g versus 10g control), and GD 6 9 ( 4.6g control verus  55.4 g at HDT). Devel NOAEL= 30 mg/ kg/ day LOAEL= 100 mg/ kg/ day based on statistically significant decreased fetal wt, resorptions and suggestive increase in malformations, 2L cleft palate & 3L pigeon chests 870.3700 & 870.3460 Inhalation Developmental in rats 00114282 ( 1979) TXR# 003413,002588,00376 4 Unacceptable; no particle size defined and inadequate description of the test conditions and data reported. Stated test doses: Test 1 0, 0.00293, 0.00635, 0.02238, Test 2; 0, 0.027, 0.060, 0.115 mg/ L Reported fetal weight decrement at 0.115 mg/ L. Reported increased incidence of runts at 0.027 mg/ L and above in one of two tests could not be verified.

Comment 40490802 Comment on 3 rabbit devel studies: The registrant did not believe that the rabbit studies suggested teratogenic effects, but agreed with the NOAEL/ LOAEL of 50/ 150 mg/ kg/ day; however, the increased post implantation loss is considered in combination with the nominally increased malformations and are considered to be suggestive of a teratogenic response at maternal toxic dose levels with NOAEL/ LOAEL of 50/ 150 mg/ kg/ day for acute exposure. Table 4.1b: Subchronic, Chronic, Developmental, Reproductive and other Toxicity Profile on Bitertanol Guideline No./ Study Type MRID No. ( year)/ Classification / Doses Results Page 15 of 58 ~ 870.3800 Reproduction and fertility effects ( Eberfeld rats, Sprague Dawley derived) readable 00114281 ( 1981) Acceptable 0, 20, 100, 500 ppm M/ F: 0, 1.0, 5.0, 25 mg/ kg/ day Parental NOAEL = 20 ppm ( F: 1.0 mg/ kg/ day) LOAEL = 100 ppm ( 5 mg/ kg/ day) based on decreased female body weight and body weight gain, premating and at 500 ppm decreased males body weight and body weight gain. Offspring: NOAEL = 20 mg/ kg/ day ( 1 mg/ kg/ day) LOAEL = 100 ppm ( 5.0 mg/ kg/ day) based on reduced f2 pup weight at birth and nominal dose related decreased at birth for f1, f3 pups and reduced lactation or viability index for f1 and f2 day 5  28 of lactation. Reproductive organs: NOAEL = 500 ppm ( 25 mg/ kg/ day) LOAEL = none ~ 870.5450 Dominant Lethal/ Mice 00025646 ( 12/ 4/ 78) 0 and 1000 mg/ kg/ day about 1/ 4 of the LD50 in mice.

Study was unacceptable because no toxicity was shown , but later upgraded to acceptable. Mice ( 50/ group) were dosed with bitertanol in a single dose at 0 or 1000 mg/ kg and on the day of dosing housed in a cage with one untreated virgin female for 4 days. This cycle was repeated for 12 females for 48 days. Fertility rate, corpora lutea counts, pre and post implantation loss and live implants were counted. Although no toxicity was demonstrated in this study, the mice were dosed at 1/ 4 of the LD50 for mice, which was probably close to a toxic dose level.

No dominant lethal effects were noted in any of the parameters tested.

~ Guideline 870.5100, Ames/ Salmonella typhimurium, reverse mutation and WP2( hcr) of Escherichia Coli reverse mutation 00114286 ( 1/ 22/ 81) Acceptable, Study# MO 01 000295 0 to 5000 g./ plate Bitertanol was tested in five strains of Salmonella [ TA 98, TA 100, TA 1535, TA 1537, TA 1538] and WP2( hcr) strain of Escherichia Coli with and without S9 at 0, 1, 5, 10, 50, 100, 500, 1000, 5000 g./ plate. Cytotoxicity was evident at 500 g./ plate and above with and without S9. A Rec assay with strains H17 and M45 with Bacillus subtilis conducted at 0, 20, 100, 200, 500, 100, 2000, 5000 g./ plate without S9 was negative. No mutagenic effects were noted with none of the tests.

Study is acceptable and negative for reverse mutation but not for Rec assay with strains H17 and M45 with Bacillus subtilis because the test was conducted only without S9..

~ Guideline 870.5100, Ames/ Salmonella typhimurium, reverse mutation 00025651 ( 1979) Mobay# 67959 Unacceptable 0 to 2500 µ g/ plate Bitertanol was test in four strains of Salmonella [ TA 98, TA 100, TA 1535, TA 1537] with S9 at 0, 4, 20, 100, 500, 2500 g./ plate and without S9 at 0 and 2500 g./ plate. Cytotoxicity was shown. No mutagenic effects were noted.

Study is unacceptable, but the lack of mutagenicity is supported by MRID# 00114286. Table 4.1b: Subchronic, Chronic, Developmental, Reproductive and other Toxicity Profile on Bitertanol Guideline No./ Study Type MRID No. ( year)/ Classification / Doses Results Page 16 of 58 ~ Guideline 870.5300, Mouse lymphoma L5178Y TK+ test ( in vitro) Acceptable 00149845 ( 2/ 23/ 83) Lab# 83/ BAG028/ 028; Mobay# 84194 0,1,5,10,15,20 g./ mL Acceptable Mouse lymphoma L5178YK+/ cells ( in vitro) were exposed 4 hours to bitertanol at 0, 1, 5, 10, 15, 20 g./ mL with and without S9 mix. Concentrations were limited by cytotoxicity.

No increased mutation frequency over controls was seen at any concentration.

~ Guideline 870.5395 Miconucleus/ mice 00025645 ( 10/ 10/ 78) Lab# 7860; Mobay# 66652; 0, 1000, 2000 mg/ kg/ day for two days Unacceptable; Only 1 sampling period & 2 required In an in vivo mouse cytogenetics test on erythrocyte micronucleous, 5 mice/ group were dosed twice, 24 hours apart with 0, 2x 1000 and 2x2000 mg/ kg/ day. Bone marrow was harvested 6 hours after the last dose. One mouse at the HDT showed ruffled coat. The study was negative, possibly due to timing in sampling. Unacceptable ( Contractor) because the bone marrow samples should have been taken between 18 hours and 24 hours instead of 6 hours after the last dose.

~ Guideline 870.5500 Bacterial DNA Damage or Repair 00114285 ( 12/ 80) Lab# 1896, 69099 0, 100, 333.3, 1000, 3300, 33300 g./ well Acceptable In a DNA repair study ( growth inhibition/ killing) in W3110( polA+) and p3478( polA) strains of Escherichia coli were exposed to bitertanol in DMSO with and without S9. Precipitation was seen in the 333.3 g./ well and above concentration, but no cytotoxicity and no growth inhibition.

There was no evidence of DNA damage in the ( preferential growth inhibition or the cell killing in the repair defective strain compared with the repair competent strain) at any concentration of bitertanol.

None Guideline Aneuploid induction in fungus 00114287 ( 7/ 78) Lab# 2034; Mobay# 80333 0, 0.1, 0.25, 0.5, 1.0, 2.5, 5.0, 10 mg/ L. Acceptable/ NG Buff X buff crosses of Sordaria brevicollis were exposed to bitertanol in DMSO at 0 to 10 mg/ L Grey X grey crosses were exposed at 0, 0.1, 0.5, 1.0, 2.5, 5.0 mg/ L. Concentrations above 2.5 mg/ L reduced fertility and ascospore maturation. Cytotoxicity was demonstrated. Induced aneuploid frequency at 5.0 mg/ L in the buff x buffy crosses, but not in the grey X grey crosses was attributed to selective maturation of black spores and buffy spores and not to aneuploid induction.

There was no evidence compound related changes in chromosome numbers in the study. Table 4.1b: Subchronic, Chronic, Developmental, Reproductive and other Toxicity Profile on Bitertanol Guideline No./ Study Type MRID No. ( year)/ Classification / Doses Results Page 17 of 58 870.7485 Metabolism & Pharmacokinetics ( Wistar rats) 00025733 ( 1979), 00129560 ( 1983) Acceptable Rats were dosed with 1 & 1000 mg/ kg in a single oral dose and multiple doses of 100 mg/ kg/ day and a single i. v. dose of 100 mg/ kg. Bitertanol was labeled in the phenyl ring only. The studies affirmed the relatively rapid absorption of [ 14C phenyl UL] BAYCOR ( Bitertanol) and that excretion was primarily via the feces ( 81 98% and 92%, respectively) over the 168 hr experimental period. Urinary excretion ( 4 11%) was secondary to fecal elimination. The i. v. dose indicated that biliary excretion was extensive. No radioactive was detected in breath samples. Tissue levels were low and indicated no bioaccumulation and the 1000 mg/ kg dose indicated metabolic saturation. There was little gender differences. Intestinal bacterial metabolism increased the proportions of neutral organic ( none acidic) extracts in rats. Parent ( both isomers) and 12 metabolites were indentified in the feces. About 50% of the radioactivity remained unextracted in the feces. Metabolites were p hydroxy Baycor ( both isomers I[ 2] & II [ 1]), m, m dihydroxy Baycor I [ 8], phydroxy m methoxy Baycor [ 5], Baycor alcohol [, p hydroxy Baycor alcohol I [ 10]& II [ 9], ether cleavage products among the conjugates and aliphatic oxidation to carboxylic acid [ 11, 12, 13, 14]. If triazole was present it was not detected. Numbered structures in brackets can be seen in the Appendix.

870.7600 Dermal absorption ( rabbit) 00150149 ( 1984) Unacceptable/ not upgradable Study only accounted for 8% and 14% of the dose applied in male and female rabbits, respectively and 92% and 86% of the applied radio label was not reported.

4.2 FQPA Hazard Considerations 4.2.1 Adequacy of the Toxicity Data Base The toxicity data base is old with deficiencies; some studies date back to 1977. However, the toxicity data base is adequate for FQPA hazard assessment of bitertanol. A combination of two rat developmental studies and three rabbit developmental studies are acceptable for developmental toxicity in the rat and rabbit. The reproduction study in the rat is acceptable, since the study showed a dose response in body weight, which is supported by the rat 90 day study at slightly higher dose levels. Only the developmental rabbit study in Chinchilla rabbits was conducted after GLP publication.

90 Day studies in the rat and dog and the chronic dog are acceptable. However, the chronic/ carcinogenicity studies were dosed at marginally adequate dose levels showing marginal toxicity. The dietary concentration of test material was not verified, such that there is no Page 18 of 58 verification of the dosage administered to the rats or mice. However, both studies were classed as acceptable, since some toxicity was demonstrated and the dose levels approached an adequate dose level. In addition, the rat and mouse were less sensitive to bitertanol toxicity than the dog and the chronic RfD was based on a lower NOAEL in a chronic dog study. A battery of adequate and negative mutagenicity studies support the lack of cancer in carcinogenicity studies.

4.2.2 Evidence of Neurotoxicity There was no evidence of neurotoxicity among the guideline studies in bitertanol toxicity data base. However, literature studies with bitertanol show limited neurotoxic effects. Bitertanol is a triazole fungicide with structural similarities with triadimefon and triadimenol. Triadimefon showed hyperactivity, but bitertanol did not in that study. Bitertanol showed an effect on a certain type of operant behavior, which may have suggested a slight hyperactivity. The difference between bitertanol and triadimefon in operant behavior is complex, but bitertanol has no effect on motor activity where as triadimefon increases motor activity [ Allen and MacPhail, 1993]. Another reference noted the difference in neurotoxic effects from bitertanol exposure and from triadimefon exposure [ Crofton, 1996]. Crofton speculated that benzene ring substitution in bitertanol for the chlorine in triadimefon sterically hindered the effect.

4.2.3 Developmental Toxicity Studies In submitted developmental toxicity studies, qualitative susceptibility was seen in the rat and rabbit at the same dose levels as maternal toxicity. A nominal increase in malformations was seen in the Long Evans rat but not in the Sprague Dawley rat. No increase in malformations was seen in the Chinchilla rabbit, but nominally increased malformations ( some of which where rare) were seen in the two Himalayan rabbit studies at the HDT. However, in all 3 rabbits studies postimplantation loss was seen at the HDT. The post implantation loss combined with nominally increased malformations in rabbits and nominally increased malformations in the rat suggests that bitertanol exposure results in terata at maternally toxic dose levels. There was no indication of increased susceptibility in the rat reproduction study.

<IP_ADDRESS> Rat Developmental Toxicity with Bitertanol [ Long Evans rats]

EXECUTIVE SUMMARY: In a developmental toxicity study ( MRID 00154936) KWG 0599 ( 96.5% a. i., lot/ batch # 16001/ 76) was administered to 20 22 female Long Evans rats/ dose by gavage at dose levels of 0, 10, 30, or 100 mg/ kg bw/ day from days 6 through 15 of gestation. On gestation day ( GD) 20, dams were sacrificed and all fetuses examined externally. Approximately one third of the fetuses were examined viscerally, and the remaining two thirds of the fetuses were examined for skeletal malformations/ variations. The total number of fetuses ( number of litters) examined was 225( 20), 238( 20), 223( 20), and 213( 19) for the 0, 10, 30, and 100 mg/ kg bw/ day groups, respectively.

Treatment with 30 or 100 mg/ kg bw/ day of KWG 0599 statistically reduced mean body weight gain of the dams over the treatment interval of GD 6 15 ( 80 and 48% of controls, respectively; p< 0.05). Dams in the mid dose group were able to recover during the 5 day post dose interval, with overall body weight gain for GDs 0 20 comparable to controls, while high dose group dams Page 19 of 58 had reduced body weight gain over the entire interval of GD 0 20 ( 84% of controls; p< 0.05). Feed consumption was not measured in this study. Gross necropsy was not conducted on the dams, so it is not known if treatment resulted in any gross lesions.

Therefore, the maternal toxicity LOAEL for KWG 0599 in rats is 30 mg/ kg bw/ day based on reduced body weight gain, and the maternal toxicity NOAEL is 10 mg/ kg bw/ day.

No treatment related, statistically significant effects on pregnancy rate, resorptions/ dam, fetuses/ litter, or fetal sex ratio were observed in the treated groups compared with the controls. An increase in the total number of resorptions in the high dose group is a consequence of one high dose dam that had complete litter resorption. The number of corpora lutea was not provided.

Treatment with 30 and 100 mg/ kg bw/ day resulted in delayed fetal growth as evidenced by a dose related decrease in fetal body weight ( 3.72 and 3.29 g, respectively vs. 3.83 g for controls; p< 0.05 for high dose) and an increase in the incidence of stunted fetuses [ fetuses( litters) affected: 8( 5) and 50( 8), respectively, vs. 1( 1) for controls]. The high dose group had an increased fetal incidence of delayed overall ossification ( 115/ 146 fetuses vs. 56/ 158 for controls) and of vertebrae ( 98/ 146 fetuses vs. 33/ 158 for controls) and sternebrae ( 39/ 146 fetuses vs. 5/ 158 for controls). Although likely an effect of treatment, the reviewer could not definitively assign significance to the delayed ossification because the study author did not report the litter incidences of these delays.

Treatment with 100 mg/ kg bw/ day resulted in an increased incidence of malformations. A total of five litters from the high dose group had fetuses with malformations compared with two litters each in the low and mid dose groups and one from the control group. Cleft palate was considered an effect of treatment with 100 mg/ kg bw/ day because it was clearly related to dose [ fetuses( litters) affected: 4( 3) vs. 0( 0) for all other groups] and was above the historical control range. Other malformations noted in the high dose group were of low incidence but were not seen in the other groups and included: a hydrocephalus, a kinked tail, rib dysplasia and synostoses, and multiple malformations of the ribs, spinal column and tail.

Therefore, the developmental toxicity LOAEL for KWG 0599 in rats is 30 mg/ kg bw/ day based on delayed fetal growth, and at 100 mg/ kg/ day nominally increased malformations and other developmental toxicity. NOAEL is 10 mg/ kg bw/ day.

The combined developmental toxicity studies in the rat ( MRID# 00154936 & 00114283) are classified Acceptable/ Guideline and do satisfy the guideline requirement for a developmental toxicity study ( OPPTS 870.3700; OECD 414) in the rat. Main deficiencies include data pertaining to the stability of KWG 0599; homogeneity, stability, and concentration analyses of the test substance in the dosing medium; and summary litter incidences of skeletal examination data. However, another unacceptable study in Sprague Dawley rats for similar reasons showed a developmental NOAEL/ LOAEL of 10/ 25 mg/ kg/ day and a maternal NOAEL/ LOAEL of 10/ 25 mg/ kg/ day ( MRID# 00114283). [ Note this study was considered acceptable in August 10, 1982 memo from Gary Bruin to Henry Jacoby ( HED File Code 13000 Tox Reviews).] Page 20 of 58 <IP_ADDRESS> Rat Developmental Toxicity with Bitertanol [ Sprague Dawley rats]

EXECUTIVE SUMMARY: In a developmental toxicity study ( MRID 00114283) KWG 0599 ( 95% a. i., lot/ batch # not provided) was administered to 25 female Sprague Dawley rats/ dose by gavage at dose levels of 0, 10, 25, or 65 mg/ kg bw/ day from days 6 through 15 of gestation. On gestation day ( GD) 20, dams were sacrificed and all fetuses examined externally. Approximately one half of the fetuses were examined viscerally, and the remaining one half of the fetuses were examined for skeletal malformations/ variations. The total numbers of fetuses examined ( number of litters) were 315( 22), 305( 23), 321( 22), and 311( 23) for the 0, 10, 25, and 65 mg/ kg bw/ day groups, respectively. Two dams from the control group died due to gavage error.

Dams treated with 25 or 65 mg/ kg bw/ day of KWG 0599 had statistically reduced mean body weight gain over the treatment interval of GD 6 15 ( 81 and 69% of controls, respectively; p< 0.05). The high dose group also had decreased absolute body weight on GD 15 ( 96% of controls; p< 0.05). Mid and high dose dams were able to recover during the 5 day post dose interval, with overall body weight gain for GDs 0 20 comparable to controls. Feed consumption was not adversely affected by treatment as measured on GD 1, 6, 15, or 20. However, it is not known if the mid and high dose groups had reduced feed consumption early in the dosing interval since feed consumption was not measured at any intermediate time points. Gross necropsy was not conducted on the dams.

Therefore, the maternal toxicity LOAEL for KWG 0599 in rats is 25 mg/ kg bw/ day based on reduced body weight gain during the dosing period, and the maternal toxicity NOAEL is 10 mg/ kg bw/ day.

No treatment related, statistically significant effects on pregnancy rate, number of corpora lutea, pre or post implantation loss, resorptions/ dam, fetuses/ litter, or fetal sex ratio were observed in the treated groups compared with the controls. No dams had complete litter resorption. No treatment related malformations or external or visceral variations were observed in any group.

A statistically significant increase in the litter incidence of the presence of a 14th rib was noted in the mid and high dose groups ( 16/ 21 and 22/ 23 litters affected, respectively, compared to 7/ 22 control litters; p< 0.05). It was not possible to assess the effect of treatment on other skeletal variations from the summary data as presented. Although the number of fetuses affected was provided, the number of litters affected was not. The reviewer did not calculate the litter incidence for the remaining skeletal variations, including the degree of ossification of the sternum, coccygeal, metacarpals, and metarsals.

Therefore, the developmental toxicity LOAEL for KWG 0599 in rats is 25 mg/ kg bw/ day based on a dose related increased incidence of a 14th rib, and the developmental toxicity NOAEL is 10 mg/ kg bw/ day.

The combined developmental toxicity studies in the rat ( MRID# 00114283 and 00154936) are Page 21 of 58 classified Acceptable/ Guideline and do satisfy the guideline requirement for a developmental toxicity study ( OPPTS 870.3700; OECD 414) in the rat. Although, major deficiencies include the following: stability of KWG 0599; homogeneity, stability, and concentration analyses of the test substance in the dosing medium, another study conducted in Germany with Long Evans rats showed similar endpoints. It should be noted that another study unacceptable for similar reasons in Long Evans rats showed malformations at 100 mg/ kg/ day and showed developmental toxicity with a NOAEL/ LOAEL of 10/ 30 mg/ kg/ day with a maternal NOAEL/ LOAEL of 10/ 30 mg/ kg/ day ( MRID# 00154936), but was considered acceptable when combined with MRID# 00114283.

There were missing fetuses in MRID# 00114283 with no explanation ( 7, 10, 7 and 0 from control, 10, 25 and 65 mg/ kg/ day dose groups, respectively, but no fetuses were missing from the HDT where the effects would be expressed if present.

<IP_ADDRESS> Rabbit Developmental Toxicity with Bitertanol [ Chinchilla rabbits]

EXECUTIVE SUMMARY: In a developmental toxicity study ( MRID 40490801) KWG 0599 ( 96.9% a. i., batch/ lot # 203 501 018) was administered to 16 female Chinchilla rabbits/ dose by oral gavage at dose levels of 0, 10, 50, or 250 mg/ kg bw/ day from days 6 through 18 of gestation. Due to excessive toxicity at the high dose, supplementary groups of 0 or 150 mg/ kg/ day were added. Does were sacrificed on GD 28 and subjected to gross necropsy. Fetuses were removed, weighed and examined for external and visceral malformations. The head was removed and examined by cross section after fixation. All carcasses were processed for skeletal evaluation. Analysis on a litter basis of data obtained during skeletal examination was presented in MRID 40829401.

At 150 and 250 mg/ kg/ day, one and two female( s) died, respectively. All remaining animals survived to scheduled sacrifice. Clinical signs of toxicity in the 250 mg/ kg/ day group included six does with hair loss in the throat region or between the hind legs, bloody toes on one animal, another doe with mucoid, yellowish flux from the vagina. Absolute body weight was similar between the treated and control groups throughout the study. The mean body weight gain and food consumption by the does at 10 mg/ kg/ day were not affected by treatment. At 50 mg/ kg/ day, no change in body weight was noted during the first three days of treatment; thereafter, the mean body weight gain was similar to that of controls. In groups treated with 250 mg/ kg/ day or 150 mg/ kg/ day, dose related effects on the body weight gain were noted. During the treatment interval, the 250 mg/ kg/ day group had a net weight loss compared to a net gain by the control group and in the 150 mg/ kg/ day group weight gain was 28% of the control level. Pre and posttreatment the mean body weight gain for all treated groups was similar to the respective control. Food consumption at 50 mg/ kg/ day was 84 88% of the control level during gestational days 6 15 or 93% of control during gestational days 6 19 of treatment and was slightly higher than control amounts for the remainder of the study. [ The same period, gestational days 6 19, showed an increase in body weight gain.] Food consumption by the 150 mg/ kg/ day group was 63 76% of controls during the first and third weeks of treatment and similar to or slightly higher than controls at other times. In the 250 mg/ kg/ day group, food consumption was 17 26% of the control level during the treatment interval, remained 53% of the control level immediately post Page 22 of 58 treatment ( GDs 19 24) and rebounded to 137% of the control level during the last four days of the study.

At necropsy, does in the 250 mg/ kg/ day group were observed with an enlarged liver which corresponded to an increase in liver weight of 36.8% compared to the control group.

The maternal toxicity LOAEL for KWG 0599 in mated female Chinchilla rabbits is 150 mg/ kg/ day, based on reduced food consumption and decreased body weight gain. The maternal toxicity NOAEL is 50 mg/ kg bw/ day.

No test article related effects were noted at cesarean section or fetal examination up to and including 50 mg/ kg/ day. In the 250 and 150 mg/ kg/ day groups post implantation losses were increased in a dose related manner ( 93.6% vs. 4.9% in controls; 16.1% vs. 5.0% in controls). The 250 mg/ kg/ day group had a total of 15 complete litter resorptions, resulting in only one litter available for evaluation. Empty implantation sites were also found in the 250 mg/ kg/ day does with complete litter resorption, but no evidence of abortion was observed. In the 150 mg/ kg/ day group, two does had complete litter resorption. Mean fetal body weight for the one live litter in the 250 mg/ kg/ day group and for the 150 mg/ kg/ day group was slightly less than that of their respective control group.

A total of 117( 15), 122( 15), 116( 15), 8( 1), 114( 15), and 73( 11) fetuses ( litters) were available for evaluation, respectively. No treatment related external or visceral malformations/ variations were seen in any fetus from any group. In the 150 and 250 mg/ kg/ day groups, the quota of incompletely or non ossified phalangeal nuclei of fore and hind limbs and calcanea was increased in a dose related manner with a shift towards a higher percentage of fetuses and litters showing reduced ossification.

The developmental toxicity LOAEL for KWG 0599 in mated female Chinchilla rabbits is 150 mg/ kg bw/ day, based on increased post implantation loss, decreased body weight and reduced ossification of the fore and hind limb digits. The developmental toxicity NOAEL is 50 mg/ kg bw/ day.

The developmental toxicity study in the rabbit is classified Acceptable/ Guideline and satisfies the guideline requirement for a developmental toxicity study ( OPPTS 870.3700; OECD 414) in Chinchilla rabbits. Some minor study deficiencies are listed in Section III. C.

<IP_ADDRESS> Rabbit Developmental Toxicity with bitertanol [ Himalayan rabbits]

EXECUTIVE SUMMARY: In a developmental toxicity study ( MRID 00114284) Bitertanol [ 96.7% a. i.; composite sample of 5 batches ( lot/ batch numbers not reported)] was administered to 12 mated female Himalayan ( CHBB: HM) rabbits/ dose by gavage in aqueous 0.5% Cremophor EL at dose levels of 0, 10, 30, or 100 mg/ kg bw/ day on gestation days ( GDs) 6 18, inclusive. On GD 29, the surviving does were sacrificed and subjected to cesarean section, and the numbers of implantation sites, male and female fetuses, and " losses," the combined weight of all placentae and of all fetuses, and the number of stunted fetuses ( weighing less than 25 g) were recorded for each Page 23 of 58 doe. Each fetus was sexed externally, examined for external malformations and malformations of the abdominal and thoracic organs and brain, and subjected to skeletal evaluation of the trunk and extremities for malformations or " slight skeletal changes."

One high dose female died on GD 29 after exhibiting diarrhea and weight loss, beginning on GD 10; the death could not be definitively attributed to treatment. Three high dose animals had hematuria ( vs. none in controls). High dose females had decreased body weight gain during the treatment interval ( 1.0 vs. 85.5 g for controls; p< 0.05) and over the entire gestation period due to decreased gravid uterine weight ( 65% of controls; n. s.). Food consumption was not measured and gross necropsy findings were not reported.

The maternal toxicity LOAEL for bitertanol in Himalayan rabbits is 100 mg/ kg bw/ day, based on clinical signs ( hematuria) and decreased body weight gain ( GD 6 18). The maternal toxicity NOAEL is 30 mg/ kg bw/ day.

At the highest dose level there was a decreased mean number of live fetuses per litter ( 5.1 vs. 6.7 for controls) due to increased postimplantation loss ( 25.0% and 1.7 " losses"/ litter vs. 7.5% and 0.6 " losses"/ litter for controls). [ Note: the " losses" were not further characterized as early/ late resorptions or dead fetuses.] No total litter resorptions were reported. Mean fetal weight and placental weight were decreased at the highest dose level ( 86% and 89% of controls, respectively; p< 0.05 for fetal weight). In the control, low, mid, and high dose groups, respectively a total of 74 ( 11), 88 ( 12), 89 ( 12), and 51 ( 10) fetuses ( and litters) were evaluated, and there were a total of 3 ( 2), 0 ( 0), 0 ( 0), and 3 ( 3) fetuses ( litters) with malformations. Two of the affected high dose fetuses had the rare malformation of epignathus or pulmonary hypoplasia and the third affected high dose fetus had aplasia of a lung or lung lobes which was also noted in the fetuses with pulmonary hypoplasia; these findings may be treatment related.

The developmental LOAEL for bitertanol in Himalayan rabbits is 100 mg/ kg bw/ day, based on decreased fetal weight, decreased placental weight, decreased live litter size, increased post implantation loss, and rare malformations. The developmental toxicity NOAEL is 30 mg/ kg bw/ day.

The developmental toxicity study in the rabbit is classified Acceptable/ Guideline and does satisfy the guideline requirement for a developmental toxicity study in the rabbit [ OPPTS 870.3700b; OECD 414]. Although, data on the homogeneity and stability of the dosing solutions were not submitted, they were stated to be confirmed. In addition, this study shows the same NOAEL/ LOAEL for maternal and developmental toxicity as MRID# 00149830; suggesting that the doses were as stated. Submitted historical control data indicated that skeletal variations in the strain of rabbit used was very low and that the study was acceptable ( TXR# 0004442). It should be noted that the study was conducted prior to the implementation of the Good Laboratory Practice standards and the current guidelines. This study also can be used as weight of evidence support of the conclusions in MRID 40490801.

<IP_ADDRESS> Rabbit Developmental Toxicity with bitertanol [ Himalayan rabbits] Page 24 of 58 EXECUTIVE SUMMARY: In a developmental toxicity study ( MRID 149830) KWG 0599 ( 93.9% a. i., mixed batch from June Oct 1978) was administered orally to 15 female Himalayan rabbits / dose in 0.5% aqueous Cremophor emulsion at dose levels of 0, 10, 30, or 100 mg/ kg bw/ day from gestation days ( GD) 6 18, inclusive. Body weight was measured on GD 0, 6 18, and 29. On GD 29, dams were sacrificed with T61 and subjected to caesarian section. Examination at sacrifice consisted of: number of implantation sites, number of male and female fetuses, number of " losses", and placental weight. Gross examination of the dams was not described. Food consumption was not reported. Fetuses were examined for external malformations and alterations. The litter weight, mean fetal weight per litter, number of stunted (< 25 grams) fetuses, and the sex of each fetus were determined. All fetuses were subjected to visceral examination. Fetuses were examined for cerebral malformations following severance of the head from the corner of the mouth to the base of the ear, and for skeletal malformations following staining with Alizarin Red S. Additional details of the methods were given in MRID 00154937 and limited historical control data were provided in MRID 40163205.

At doses up to and including 30 mg/ kg/ day there were no treatment related effects on maternal survival, clinical signs, body weight, food consumption, or caesarean parameters. However, at 100 mg KWG 0599 / kg bw/ day, the dams reportedly ate less, although no data are presented to confirm the magnitude of effect. Further, two dams bled occasionally from the vagina. Absolute body weight was similar between the treated and control groups throughout the study. The mean weight gain of the control group and the treated groups did not differ significantly from each other, however, weight gain by the high dose group during the treatment interval was 60% of the control group level. All groups including controls lost weight during the first three days of treatment with the greatest magnitude in the high dose group ( 55.6 g vs  4.6 g for controls). The evidence for maternal toxicity is an unsupported statement by the authors that rabbits at 100 mg/ kg/ day ate less and past fewer stools than other groups, one abortion occurred, vaginal bleeding was reported for two rabbits during the study and the body weight gain decrement during GD 6 9. The evidence for maternal toxicity is marginal at best.

The tentative maternal toxicity LOAEL for KWG 0599 in Himalayan rabbits is 100 mg/ kg bw/ day based on reduced body weight gain. The tentative maternal toxicity NOAEL is 30 mg/ kg/ day.

At doses up to and including 30 mg/ kg/ day, there were no treatment related effects on developmental parameters. The high dose of 100 mg/ kg/ day was embryotoxic and teratogenic; dams in this dose group had a significantly higher resorption rate ( postimplantation loss: 64.2% vs 21.0% in controls) due to seven high dose animals with complete liter loss [ one abortion and 6 litters with complete resorption. In addition, one high dose doe aborted on GD 23. The total number of fetuses ( litters) evaluated was 81 ( 14) control, 90 ( 15) low dose, 86 ( 15) mid dose, and 34 ( 8) high dose. Mean fetal body weight in the high dose group was significantly lower than controls ( 34.94 g vs 39.38 g for controls). Finally, there was a greater incidence of fetal malformations in the 100 mg/ kg dose group compared to controls ( 1 fetus in the control group had one type of malformation; 6 fetuses from 4 high dose litters had one or two types of malformations). Fetal malformations in the 100 mg/ kg/ day dose group included cleft palate ( 2 fetuses; 2 litters), " pigeon chest" ( 4 fetuses; 2 litters), arthrogryposis ( 1 fetus), abnormal posture of the hind extremities ( 1 fetus), and rib and spinal column adhesions ( 1 fetus). In the control Page 25 of 58 group, only one fetus was observed with a malformation ( arthrogryposis) and 2 fetuses from 2 litters in the 10 mg/ kg/ day dose group had malformations ( rib and spinal column adhesions; umbilical hernia and fusion of ribs); no malformations were observed in the 30 mg/ kg/ day dose group.

The tentative developmental toxicity LOAEL for KWG 0599 in Himalayan rabbits is 100 mg/ kg bw/ day, based on increased resorption rate, decreased body weight, and increased incidence of fetal malformations. The tentative developmental toxicity NOAEL is 30 mg/ kg bw/ day.

The developmental toxicity study in the rabbit is classified Unacceptable/ Guideline and does not satisfy the guideline requirement for a developmental toxicity study ( OPPTS 870.3700; OECD 414) in rabbits. Major deficiencies include lack of analyses of homogeneity, stability, and concentration of the dosing solutions. The lack of analysis of dosing concentrations could be mitigated by the similar NOAEL/ LOAEL to MRID# 00114284, but the omission of data requested that could verify some of the effects, prevents this study form being acceptable. The data requested should be submitted. For a list of other study deficiencies, refer to Section III. C. This study can be used as weight of evidence support of the conclusions in MRID 40490801. [ This study was unacceptable in TXR# 0006528, but concluded that fetal toxicity in the form of fetal weight decrement, fetal lose/ dam and decreased number of litters was shown at 100 mg/ kg/ day.]

4.2.4 Reproductive Toxicity Study Bitertanol EXECUTIVE SUMMARY: In a 3 generation reproduction study ( MRID 00114281), bitertanol [ 95.0% a. i.; batch nos. 16012/ 77 and 16001 16004/ 78 ( mixed sample)] was administered to 10 male and 20 female FB 30 ( Elberfeld)( Sprague Dawley derived) rats/ group in the diet at concentrations of 0, 20, 100, and 500 ppm ( equivalent to 0, 1, 5, and 25 mg/ kg bw/ day using a food factor of 0.05 for the rat). Parental animals were administered the treated or control diets for 56 days ( F 0 ) or 70 days ( F 1 and F 2 ) prior to mating, throughout mating, gestation, and lactation, and until necropsy. Two litters ( A and B) were produced by each generation, and the parental animals for the next generation were selected from the B litters. In the absence of analytical data for the diet formulations and lack of food consumption data, the actual dosages to the study animals are unknown; therefore the LOAELs and NOAELs set below are tentative ones.

There was no treatment related effect on mortality, and no treatment related clinical signs were reported. High dose males and females of all three generations consistently had decreased body weight gain during the first 2 3 weeks of premating ( F 0 , F 1 , and F 2 males: 45 89%, 81 83%, and 71 80% of controls, respectively; F 0 , F 1 , and F 2 females: 44 81%, 75%, and 65 77%). The significant decreases in absolute body weight persisted throughout premating, but tended to decrease in magnitude over time ( F 0 , F 1 , and F 2 males: 76 91%, 69 88%, and 66 92% of controls; F 0 , F 1 , and F 2 females: 77 91%, 71 89%, and 68 87%). F 1 mid dose females had marginally decreased absolute body weight during weeks 9 14 ( 91 92%) after gaining 3 4 g less per week than controls during weeks 5 9 and had decreased weight gain during weeks 9 11 ( 87% of controls). F 2 mid dose females also had significant decreases in absolute body weight throughout premating ( 84 89%; p< 0.01) along with slightly decreased body weight gain during most Page 26 of 58 intervals. No treatment related gross changes were noted at necropsy of the F 2 adults, and there were no treatment related effects on organ weight. Microscopic examination of tissues from F 2 adults revealed bile duct proliferation at increasing severity in mid and high dose males and at increasing incidence and severity in high dose females. In males, this finding was seen in 2/ 5 controls, 1/ 5 low, 1/ 5 mid, and 2/ 5 high dose males, with average grades of 1.0, 1.0, 2.0, and 2.5, respectively. In females this finding was seen in 1/ 5 and 4/ 5 mid and high dose animals, respectively ( vs. 0/ 5 controls) with average grades of 1.0 and 2.3, respectively. However, since microscopic evaluation was limited to 5 F 2 animals/ sex/ dose, these data are of limited usefulness and were not used to set toxicity levels.

The parental systemic LOAEL for bitertanol in FB 30 ( Elberfeld) rats is 100 ppm for females ( approximately 5 mg/ kg bw/ day) and 500 ppm for males ( approximately 25 mg/ kg bw/ day), based on decreased body weight and body weight gain during premating. The parental systemic NOAEL is 20 ppm for females ( approximately 1 mg/ kg bw/ day) and 100 ppm for males ( approximately 5 mg/ kg bw/ day).

Treatment related effects on offspring body weight and body weight gain were noted in high dose pups from all generations and litters, in the mid dose F 2a and F 2b pups, and the mid dose F 3a pups. The high dose F 3b pups had significantly decreased mean body weight beginning on LD 5, postcull and continuing through the remainder of lactation ( 79 87% of controls), while all of the other high dose pups had significantly decreased mean weight at birth and throughout lactation ( F 1a : 75 80%; F 1b : 70 90%; F 2a : 76 86%; F 2b : 62 76%; F 3a : 84 89% of controls). In high dose pups, the greatest effect on body weight gain was during LD 0 5; cumulative weight gain during the LD 0 28 interval was also decreased ( F 1a and F 1b : 78% and 68%; F 2a and F 2b : 80% and 64%; F 3a and F 3b : 88% and 78% of controls, respectively). Mid dose F 2 pups had significantly decreased body weight throughout lactation ( F 2a : 78 93%; F 2b : 71 84%); the F 2a pups mainly had decreased weight gain during LD 0 5 ( 60%), with absolute body weight showing evidence of recovery by LD 28, whereas the F 2b pups had decreased weight gain throughout lactation ( LD 0 5: 57%; LD 5 28: 79%) and did not show recovery. The mid dose F 3a pups had decreased weight gain at the beginning of lactation ( 82%); absolute pup weight was not affected until LDs 7 and 14 ( 86 88% of controls), and recovery had occurred by LD 21. Additional evidence of offspring toxicity seen at the highest dose level included decreased pup survival ( decreased viability index and/ or lactation index) in all high dose litters of all generations. Correspondingly, decreased litter size occurred in the high dose F 1 A litters on LDs 0 and 5, pre cull ( 81% and 73% of controls, respectively), and in the high dose F 3 A litters throughout lactation ( LD 0: 77%; LD 5, pre cull: 67%; LD 28: 69% of controls).

The offspring LOAEL for bitertanol in FB 30 ( Elberfeld) rats is 100 ppm ( approximately 5 mg/ kg bw/ day), based on decreased body weight and body weight gain. The offspring NOAEL is 20 ppm( approximately 1 mg/ kg bw/ day).

No clear treatment related effects on reproductive performance ( fertility and gestation indices) were seen in any generation or litter. Ovarian and testicular weights of the F 2 animals were not affected by treatment, and no gross changes of the testes, epididymides, ovaries or uteri of the F 2 adults and F 3b weanlings ( 10/ sex/ group) were reported. Histopathological evaluation of testes and ovaries from 5 F 2 animals/ sex/ group and testes, ovaries, and uteruses from 10 F 3b Page 27 of 58 weanlings/ sex/ group did not reveal any clear treatment related changes.

Under the conditions of this study, the reproductive LOAEL for bitertanol in FB 30 ( Elberfeld) rats is not identified, and the reproductive NOAEL is greater than or equal to 500 ppm ( approximately 25 mg/ kg bw/ day).

This study is Acceptable/ Guideline and does satisfy the guideline requirement for a 2 generation reproductive study [ OPPTS 870.3800; OECD 416] in rats. Although there were no analyses for concentration, homogeneity and stability of the test material in the diet, the study is acceptable for the following reasons: ( 1) A dose related response was noted in several parameters, indicating that the animals were dosed. ( 2) The body weight decrement data is supported by an acceptable 90 day study in a similar strain of rat with a NOAEL/ LOAEL = 3.6/ 18.6 mg/ kg/ day ( MRID# 00157853) and an acceptable 90 day study in dogs with a NOAEL / LOAEL of 1/ 5 mg/ kg/ day ( MRID# 00025648).

Although there were a number of other deficiencies in the conduct of this study [ see deficiency section], the study was conducted prior to the implementation of the guidelines and these deficiencies did not adversely affect the quality of the data that was attained. However, rotating each female with 3 males without an evaluation of each mating as was done in this study is not a recommended practice.

4.2.5 Additional Information from Literature Sources Several papers considered aspects of bitertanol toxicity were found in the literature. FAO and WHO working groups [ 1983] & [ 1987] reviewed toxicity data of bitertanol. In general the review of the toxicity data agreed with the results in the DERs by EPA, including the rabbit dermal study, the rat chronic study, the chronic dog study and the metabolism study in rats. These references indicated the1000 mg/ kg dose level resulted in saturation of the metabolic system, which showed reduced absorption and increased unmetabolized parent material in the feces.

Another FAO and WHO working group paper [ 1998] reviewed more completely the toxicity and metabolism data on bitertanol than did the 1983 paper. The review of the toxicity data confirmed the interpretation of similar studies by EPA. Metabolism studies of bitertanol showed that 84% of the orally administered dose was rapidly absorbed, metabolized and excreted through the bile into the feces. The terminal phase of elimination from the plasma showed a ½ life was 26 hours. Parent compound was not detected in bile or urine. Parent was found only in the feces of which 15% of the administered dose probably represented unabsorbed material. Radiolabel was excreted in the urine [ 4% 11%] and the remaining material in the feces. Para hydroxy bitertanol was the only metabolite identified in the bile. Smaller amounts of para hydroxybitertanol acid, parabitertanol alcohol and bitertanol acid were identified. Ring dihydroxylation, aryl 0 methylation, ether cleavage products, and aliphatic hydroxylation and oxidation to acid with gluronide and sulfate conjugates were tentatively identified.

The 1998 reference also reviewed studies of central nervous system. Mice administered single doses of bitertanol at 0.075, 0.6 or 4.8 mg/ kg. The results showed no significant effects with Page 28 of 58 potentiation of anesthesia, stimulation of spontaneous motility, open field test, reserpine ptosis, potentiation of amphetamine effects or antagonism to reserpine ptosis. Mice administered single oral doses of 0.02, 0.2, 2, 20 or 200 mg/ kg showed significantly increased spontaneous motor activity at 200 mg/ kg only. Rats showed increased spontaneous motor activity at single dose of 200 mg/ kg, but not at 100 mg/ kg orally or intraparitoneally.

A radiolabeled 50% formulation of bitertanol administered dermally to rabbits showed a 10% dermal penetration. The 1998 reference cited Hixson, 1984. [ Study was classified by OPP as unacceptable because only one dose level was used and the study did not account for 86 to 92% of radio label applied.]

Four references were found relative to the developmental toxicity of bitertanol. Schardien ( 1993) referred to a short note by Vergieva ( 1990) in stating that bitertanol resulted in microacardia and acaudia and in rare cases exophtalamus, hypgnathia and cleft palate. The short note stated that a clear dose effect relationship was established from single doses at 100, 500 and 1000 mg/ kg on gestational day 9, 10, 11 or 13. The two other developmental toxicity studies in the rat were TSCA Section 8( e) submissions already discussed under MRID# 00154936 ( studied by gavage dosing) and 00114282 ( studied through inhalation dosing). The latter study was inadequate because no particle sizes were discussed. The study ( MRID# 00154936), which used doses by gavage at 0, 10, 30 or 100 mg/ kg/ day, showed maternal weight reduction and stunted fetal growth at NOAEL/ LOAEL of 10/ 30 mg/ kg/ day. Bone anomalies were statistically increased in a treatment related manner at 100 mg/ kg/ day when sternebrae ( delayed ossification) and vertebrae ( dumb bell) anomalies were added together.

Lewalther and Korallus ( 1986) advocated testing for exposure to bitertanol by detecting 4 hydroxyphenylphenol aducts to erythrocytes.

Allen and MacPhail [ 1993] noted that Long Evan rats showed increases in operant responding behavior, but not motor activity. Operant Behavior: Operant chambers using milk delivery for reinforcement and 3 multicolored cue lights centered a lever on the front panel and a house light was mounted at the top of the panel above a speaker. Below the speaker was a Sonalert device that was briefly activated following a response. The rats were trained to consume milk from the dipper and then to press the lever. After all the rats performed on a FR 2 schedule for one session, the multiple FI 1 min F1 5 min schedule ( with a 60 s limited hold in each component) was initiated. Discriminative stimuli were illumination of either the house light ( F1 1 min) or the cue lights ( F1 5 min). Motor Activity: Motor activity chambers were used with a platform that housed 5X8 matrix of photodetectors that were illuminated by a single overhead incandescent lamp. Each movement that occluded a photodetector was recorded as a horizontal activity count ( 6 rats/ group).

Operant behavior results were recorded as a percentage of the control response rate. Bitertanol was administered orally in a single dose at 0, 10, 30, 56, 100 or 300 mg/ kg. Bitertanol showed no effect on overall response rates or index of curvature ( IOC) at lowest dose[ 10 mg/ kg], but response rates increased for both F1 components, but were greater in F1 5 min than the F1 1 min component at the three intermediate doses [ 30, 56, and 100 mg/ kg] and then fell to control levels at 300 mg/ kg. Unlike triadimefon, bitertanol administration had no effect on motor activity in Page 29 of 58 these rats. These response patterning were similar to triademifon and other stimulants, and dissimilar from those reported for most other pesticides. Bitertanol response patterning in the operant behavior tests resembled the effects of triadimefon, but the response was not as great. The data were reported as a percentage of control and did not clearly state the number of animals used/ group to test for operant behavior.

Crofton [ 1996] noted that the triazole pesticide triadimefon exposure affects the CNS catecholamines and introduces a transient hyperactivity and sterotyped behaviors in rats. Of the 16 triazole pesticides studied for hyperactivity in a figure eight maze with photodetectors, only triadimefon and triadimenol showed hyperactivity when dosed acutely. Crofton suggested that the SAR for triazole induced hyperactivity is fairly rigid and requires the ether linkage between the phenyl group and the aliphatic group for activity; and the lack of effect of bitertanol, which has this ether linkage, was the result of stearic hindrance from the benzene ring substituted for a chlorine of triadimefon and triadimenol.

The 1998 FAO and WHO reference indicated the acute toxicity of metabolites of bitertanol. A ketone metabolite of bitertanol, 1( triazol 1 yl) 1( 4' phenyphenoxy) 3,3 dimethylbutan 2 one, induced signs of effects on the CNS, with initial sedation and respiratory disturbances and later stimulation. The highest oral dose technically administrable was1750 mg/ kg, which caused no deaths. A metabolite not seen in rat studies, bitertanol benzoic acid at an oral acute dose of 5000 mg/ kg showed no toxicity. 1,2,4 triazole given orally in an acute dose showed an LD50 of 1600 mg/ kg. When administered dermally the acute LD50 was 4200 mg/ kg. 1,2,4 triazole was strongly irritating to the mucus membranes of rabbit eyes. In a 90 day subchronic study at 0, 100, 500, or 2500 ppm, 1,2,4 triazole administration orally resulted in temporary reduction in food intake, body weight and transient palmospasms in isolated animals. Significant decreases in hemoglobin, hematocrit, mean corpuscular volume and mean corpuscular hemoglobin in male rats at 2500 ppm. Fat accumulation was seen in the 3/ 15 male livers at 2500 ppm. The NOAEL was 500 ppm ( 38 mg/ kg/ day). In developmental toxicity in the Wistar rat, the triazole metabolite administration resulted in reduced fetal weight, increased resorption, and malformations including cleft palate and malformed extremities at maternal toxic dose levels. The NOAEL was 30 mg/ kg/ day, which was higher than the NOAEL of 10 mg/ kg/ day with bitertanol in Long Evans and Sprague Dawley rats at maternally toxic doses.

4.2.6 Pre and/ or Postnatal Toxicity <IP_ADDRESS> Determination of Susceptibility Qualitative susceptibility was seen in two developmental toxicity studies [ 1982 1983] in Himalayan rabbits and qualitative susceptibility was seen in one developmental toxicity study in Chinchilla rabbits [ MRID# 40490801, 1987] that showed increased resorptions at dose levels causing slight maternal weight gain decrement. These studies showed no increased quantitative susceptibility. The two rat developmental toxicity studies showed stunted fetuses and increased incidence of 14th rib at maternally toxic dose levels, and at higher dose levels nominally increased incidence of malformations in one rat study, but no increased susceptibility. No offspring pre or postnatal susceptibility was seen in a 3 generation reproduction study. Page 30 of 58 <IP_ADDRESS> Degree of Concern Analysis and Residual Uncertainties for Pre and/ or Post natal Susceptibility There was evidence of malformations at high doses in two developmental toxicity studies in rats, but no quantitative susceptibility and NOAELs were clearly defined. In rabbits there were three developmental studies which, when combined, clearly delineated developmental toxicity in the rabbit. Although there were some deficiencies in the rat reproduction study, the study is considered acceptable and indicates that there are adequately defined NOAELs and no evidence of susceptibility. There is no residual uncertainty with respect to pre and post natal toxicity based on the submitted developmental and reproduction studies.

4.3 Recommendation for a Developmental Neurotoxicity Study 4.3.1 Evidence that supports requiring a Developmental Neurotoxicity study The literature refers to single dose studies showing effects on operant behavior ( Allen and MacPhail, 1993). The effect on operant behavior was distinguishable from and less than with triadimefon and triadimenol and unlike the latter two pesticides showed no effects on motor activity. However, decreased motor activity was seen in mouse and rat single dose studies at 200 mg/ kg ( FAO and WHO working group, 1998). No multi dose studies related to neurotoxicity have been seen with bitertanol. One fetus in one litter showed hydrocephalus in a developmental toxicity study.

4.3.2 Evidence that supports not requiring a Developmental Neurotoxicity study No neurotoxicity or CNS developmental toxicity was seen in the developmental toxicity studies in the rat or rabbit or in the study on reproduction. No neurotoxicity was seen in the subchronic or chronic guideline studies in rats, mice or dogs.

<IP_ADDRESS> Rationale for the UFDB A DNT study is not recommended at this time. A data base uncertainty factor of 10X was assessed due to the uncertainty caused by the lack of multi dose neurotoxicity studies with bitertanol. Acute neurotoxicity battery and 90 day neurotoxicity studies are necessary to resolve the conflicting literature reports and to determine NOAELs from guideline studies. A DNT study recommendation is held in reserve pending review of the acute and 90 day neurotoxicity studies.

4.4 Hazard Identification and Toxicity Endpoint Selection 4.4.1 Acute Reference Dose ( aRfD)  Females age 13 49 Selected Study: Developmental toxicity in rabbits ( MRID# 40490801). GDL# 870.3500. For an Executive Summary see Section <IP_ADDRESS> Page 31 of 58 Dose and Endpoint for Establishing an aRfD: NOAEL is 50 mg/ kg/ day. LOAEL is 150 mg/ kg/ day based on developmental effects of nominal increased incidence of malformations , increased resorptions, post implantation loss, decreased fetal weight and delayed ossification.

Uncertainty Factor ( UF): 1000X. This includes 10X for interspecies extrapolation, 10x for intraspecies variation, and a 10X database uncertainty factor the lack of neurotoxicity studies.

Comments about the Study/ Endpoint/ uncertainty Factor: The nominal increase in malformations, resorptions and post implantation loss is presumed to be a single dose effect. The developmental endpoint is appropriate for females 13 49 and is of the appropriate duration. The endpoint is supported by a two rat studies with bitertanol with NOAEL/ LOAEL of 10/ 25 & 10/ 30 mg/ kg/ day.

4.4.2 Acute Reference Dose ( aRfD)  General Population An appropriate acute endpoint attributable to a single dose was not available in the toxicity data base including the developmental toxicity studies.

4.4.3 Chronic Reference Dose ( cRfD)

Selected Study: The chronic study in dogs ( MRIDs 00157466 and 00157465 and supplemental MRIDs 40163203, 40163202, 40163201, and 40186401) GDL: 870.4100 EXECUTIVE SUMMARY: In chronic toxicity studies ( MRIDs 00157466 and 00157465 and supplemental MRIDs 40163203, 40163202, 40163201, and 40186401), KWG 0599 [ Bitertanol; MRID 00157466: 1616001/ 78 ( 97.3% a. i.); Composite of Batch Nos. 16012/ 77 and 16001 16004/ 78 ( 95% a. i.); MRID 11457465: Composite of Batch Nos. 816164054, 0816164056, 816164067, 816164079, and 816164083 ( 96.7% a. i.); Composite of Batch Nos. 233296089, 233296233, 233296296, 233296989, and 233296992 ( 96.3% a. i.)] was administered in the diet to beagle dogs. In MRID 00157466, the test material was provided to groups of four male and four female dogs at concentrations of 0, 10, 40, or 160 ppm ( equivalent to 0, 2.11, 8.18, and 32.24 mg/ kg/ day) for 104 weeks. In MRID 00157465 the test material was provided in the diet to groups of six male and six female dogs at concentrations of 0, 3, or 25 ppm ( equivalent to 0, 0.78, and 7.14 mg/ kg/ day) for 52 weeks and to a group of six male and six female dogs at 200 ppm ( equivalent to 50.93 mg/ kg/ day) for 86 weeks.

Treatment with the test material had no effect on mortality, body weight, reflexes, pulse, food intake, hematological parameters, or urinalysis parameters. Treatment did induce slight increases in the enzyme activities of ALP and ALT, enzymes associated with the liver. No increases were found in the liver activity of total cytochrome P 450 or N demethylase. In MRID 00157466, slight increases in the absolute and/ or relative to body weight of the liver of high dose dogs were observed while in MRID 00157465, the absolute and relative adrenal weights of 200 ppm male dogs were increased. In both studies, treatment with 160 ppm for two years, or 200 ppm for 86 weeks increased the incidence ( 3/ 8 at 160 ppm and 3/ 11 at 200 ppm) of corneal opacity. These liver, ocular, and adrenal effects were considered KWG 0599 related. Page 32 of 58 In MRID 00157466, minimal to moderate adrenal vacuolation was found in all 160 ppm dogs, 7/ 8 40 ppm dogs had very minimal to moderate adrenal vacuolation and 3/ 8 10 ppm dogs had very minimal adrenal vacuolation. Adrenal vacuolation was not observed in control dogs. In MRID 00157465, all male and female dogs treated with 200 ppm test material developed slight to medium adrenal vacuolation by 86 weeks of treatment. The vacuoles were characterized as large amyloid droplets in the epithelia of the inner cortical layer close to the medulla. Medium to severe thymic involution was found in 4/ 6 200 ppm male and 2/ 6 200 ppm female dogs. Thymic atrophy was not found in control, 3 ppm or 25 ppm male and female dogs and was not reported in MRID 00157466.

The 86 weeks study at 200 ppm [ MRID 00157465] confirmed the slight liver effects at 160 ppm in the 104 week study [ MRID# 00157466] and the adrenal effects in the 104 week study at 40 ppm. The adrenal vacuolation at 10 ppm was minimal and of doubtful toxicological significance. However, the lack of adrenal vacuolation at 25 ppm in a 52 week study can not be used as a NOAEL for a 2 year study. Therefore the NOAEL for the 2 year study is considered to be 10 ppm.

The LOAEL for MRID# 00157466 & 157465 with bitertanol in male and female dogs is 40 ppm ( 8.18 mg/ kg/ day) based on adrenal vacuolation and at 160 and 200 ppm, adrenal vacuolation, eye and liver effects. The NOAEL is 10 ppm ( 2.11 mg/ kg/ day).

This chronic study in the dog is Acceptable/ Guideline and satisfies the guideline requirement for a chronic oral study [ OPPTS 870.4100, OECD 452] in the dog.

Dose and Endpoint for Establishing a cRfD: NOAEL is 2.11 mg/ kg/ day. The LOAEL is 8.18 mg/ kg/ day for male/ female based on adrenal vacuolation.

Uncertainty Factor ( UF): 1000X. This includes 10X for interspecies extrapolation, 10X for intraspecies variation, and a 10X database uncertainty factor for the lack of neurotoxicity studies.

Comments about the Study/ Endpoint/ uncertainty Factor: The study is the right duration. The dose levels in the chronic dog study were considered more acceptable than those in the 90 day dog or the reproduction study. Although, the 90 day dog study and the 3 generation study on reproduction showed a NOAEL of 1 mg/ k/ day, the 90 day study was not chosen due to spacing of the doses, and the reproduction study was not chosen because the dose levels were calculated from standard tables and not measured.

4.4.4 Incidental Oral Exposure ( Short Term, 1 30 days), and Intermediate Term, 1 month 6 months)

These endpoints are not required since there are no residential exposures.

4.4.5 Dermal Absorption A 1998 FAO and WHO report stated that dermal absorption was about 10% [ MRID# 00150149]. However, OPP reviewed the data and found the study unacceptable because it did not account Page 33 of 58 for 86% to92% of the radio label applied to the test animals.

4.4.6 Dermal Exposure ( Short, Intermediate and Long Term).

These endpoints are not required for an import tolerance.

4.4.7 Inhalation Exposure ( Short Term, 1 30 days, Intermediate Term, 30 day  6 months or Long Term, > 6 months)

This parameter is unnecessary because there is no inhalation exposure in the United States.

4.4.8 Margins of Exposure Not applicable.

Occupational exposure: An occupational exposure assessment is not required for an import tolerance.

Residential exposure: There are no residential exposures for this risk assessment.

4.4.9 Recommendation for Aggregate Exposure Risk Assessments Not applicable.

4.4.10 Classification of Carcinogenic Potential Bitertanol has not been reviewed by the Cancer Assessment Review Committee [ CARC], however, no dose related carcinogenicity was seen in the rat [ MRID# 00114279] or mouse [ MRID# 00114280] carcinogenicity studies.

The carcinogenicity studies in the rat and mouse were conducted at a marginally toxic dose levels and neither study analyzed test material concentration in the feed. However, both studies showed some toxicity at the HDT, where no treatment related carcinogenicity was seen. The toxicity seen at the HDT in both studies is evidence that the animals were dosed and the effects in rats are consistent with subchronic studies in rats where doses were analyzed. Although, both carcinogenicity studies could have used higher doses, both studies were classified as acceptable, because, ( 1) Bitertanol appears to be less toxic in the rat than in the dog, which was used to set the RfD, ( 2) No treatment related carcinogenicity were seen in the rat or the mouse studies at marginally toxic dose levels, ( 3) Bitertanol is negative in a battery of mutagenicity studies, ( 4) Bitertanol is used on import bananas only, thus exposure is limited to residues on bananas. ( 5) Both studies were listed as acceptable in 1988.

<IP_ADDRESS> Combined Chronic Toxicity/ Carcinogenicity Study in Rats ( MRID# 00114279) GDL# 870.4300 EXECUTIVE SUMMARY: In a combined chronic toxicity/ carcinogenicity study ( MRID Page 34 of 58 00114279), KWG 0599 ( bitertanol; 94 95%; batch/ lot # 1616001/ 78 and 16012/ 77 & 16001 16004/ 78 composite) was administered in feed to groups of 50 male and 50 female Wister TNO/ W 74 rats at concentrations of 0, 20, 100, or 500 ppm for 2 years. The equivalent doses were 0, 1.01, 4.89, and 25.54 mg/ kg bw/ day, respectively, for males and 0, 1.32, 6.55, and 33.53 mg/ kg bw/ day, respectively, for females. An interim sacrifice group was not included in this study.

No treatment related effect was observed on clinical signs or mortality. Adequate numbers of male and female rats were alive at the end of the study for evaluation of late developing lesions. High dose male and female rats weighed 11% and 9% less, respectively, than controls at the end of the first week of treatment and body weight was within 10% of control weight for the remaining weeks except for a 12% decrease in the high dose female group at week 91. High dose males gained 42% less weight than controls during the first week of treatment, 8% less during the first year, and 77% less during the second year resulting in an 11% decrease in weight gain for the entire study. High dose females gained 41% less weight than controls during the first week of treatment, 15% less during the first year, an amount similar to that of controls during the second year, resulting in a 13% decrease in weight gain for the entire study. Body weight and weight gain were similar among the low dose and mid dose and control groups. Weekly food consumption by high dose male and female rats fluctuated considerably throughout the study, but total food consumption/ group and average daily food consumption/ rat were similar to those of the control group. Food efficiency calculated for the entire study was reduced by 9% in high dose male and female rats, suggesting that the reduced weight gain was likely due to a slightly toxic effect of the test material.

Statistically significant decreases were seen at the HDT in hematological and changes in clinical chemistry parameters, which were within the normal range. Thus, no treatment related effects were observed on hematologic, clinical chemistry, or urinalysis parameters. Postmortem examination showed no treatment related effects on the incidences of gross or microscopic lesions. It is noted that the statistically significant lung alveolar macrophage aggregation in males and females at 500 ppm may have resulted from inspiration of dietary dust. The tracheitis/ peritracheitis at the two highest doses may be related to test material irritation from diet consumption.

The lowest observed adverse effect level ( LOAEL) for KWG 0599 in rats is 500 ppm ( 25.54 mg/ kg/ day for males and 33.53 mg/ kg/ day for females) based on decreased weight gain and food efficiency. The no observed adverse effect level ( NOAEL) is 100 ppm ( 4.89 mg/ kg/ day for males and 6.55 mg/ kg/ day for females).

At the doses tested, there was no treatment related increase in tumor incidence in male or female rats receiving any dose of the test material when compared to controls. Although the high dose caused a notable decrease in weight gain in male and female rats during either the first or second year of the study, these animals could have tolerated a higher dose, because absolute body weight remained within 10% of that of the control group and no increase in mortality, adverse clinical pathologic effects, or histopathologic lesions were observed that could be attributed to treatment with the test material. Page 35 of 58 This chronic/ carcinogenicity study in the rat is Acceptable/ Guideline and does satisfy the guideline requirement for a chronic toxicity/ carcinogenicity study OPPTS 870.4300); OECD 453] in the rat. Although analyses of concentration of test material in the diet was not done, the study showed sufficient toxicity to show that the animals were dosed, the weight of evidence would suggest that this study can be classified as acceptable. ( 1) The effects noted in this study are consistent with effects in an acceptable 90 day study in Sprague Dawley rats and a reproduction study. ( 2) A battery of mutagenicity studies are all negative. ( 3) The HDT showed some toxicity and was close to an adequate dose. ( 4) There was no indication of dose related increased tumor incidence at any dose including the top dose. ( 5) An acceptable Chronic Dog study showed a lower NOAEL. ( 6) The study is used to support a limited exposure on imported bananas only. ( 7) The study is listed as acceptable in TXR# 0002588. Additional deficiencies, which do not negate the study are listed in Section IIIC of this DER.

<IP_ADDRESS> Carcinogenicity Study in Mice ( MRID# 00114280) GDL# 870.4200 EXECUTIVE SUMMARY: In a 24 month oral carcinogenicity study ( MRID 00114280), KWG 0599 ( 97.3% a. i., Batch # s 1616001/ 78, 16012/ 77, and 16001  16004/ 78) was administered to a total of 50 SPF CF 1 / W 74 ® mice/ sex/ dose in the diet at concentrations of 0, 20, 100, or 500 ppm ( equivalent to 0, 5, 25, or 129 mg/ kg bw/ day for males and 0, 7, 37, or 183 mg/ kg bw/ day for females).

There were no significant treatment related effects on mortality, clinical signs, food intake, or hematology parameters. At 500 ppm, mean weekly body weights of males were within 95% of controls at most weighing periods throughout the study, and of females were within 95% of controls during the first 12 months. The final body weights of the 500 ppm males and females were 97% and 92% of controls, respectively, and the overall body weight gains were 92% and 79% of controls, respectively. The overall food efficiency in males was decreased by 21% in females at 500 ppm, suggesting a treatment related decrease in body weight in 500 ppm females. Although the body weight decrement was slight in males and females, it was consistently decreased in the 500 ppm group from the controls, 20 and 100 ppm groups throughout the study, suggesting that the 500 ppm groups were affected by treatment and that the 500 ppm dose level was close to a toxic dose level even in males. Other studies with bitertanol have shown toxicity from histopathological findings, but only marginal body weight decrement.

The mean alkaline phosphatase activity was statistically significantly increased at 100 and/ or 500 ppm in males and females after 12 months only. The mean glutamate pyruvate transaminase ( GPT) activity was statistically increased in all male dose groups after 12 months, and in females at 500 ppm after 24 months. The enzyme activity increases were not dose dependent and largely due to outliers with very high levels of enzyme activity. The mean absolute and relative ( to body weight) liver weights were increased at 500 ppm in males ( 16% and 19%, respectively) and females ( 60% and 69%, respectively) although only the latter differed statistically from the controls. An increased incidence of greenish liver discoloration occurred in 500 ppm females, being seen only in those that survived to terminal sacrifice. Microscopic examination revealed that females had an increased incidence of liver altered cell foci ( basophilia, eosinophilia, vacuolation) that was not clearly dose related ( 16%, 40%, 26%, and 54% at 0, 20, 100, and 500 ppm, respectively). The lack of histopathological findings indicates that the liver weight increase Page 36 of 58 in 500 ppm females was a non specific adaptive response to treatment with a xenobiotic. Significant increases found in both sexes in blood urea concentration and in the incidence of kidney cortical cysts were not dose related, and were not considered treatment related.

A LOAEL was 500 ppm ( M/ F: 129/ 183 mg/ kg/ day) based marginal body weight decrement in males and females ( not statistically significant) throughout the study, and other statistically significant marginal effects, such as eosinphilic foci in livers of females. Increased liver wt & GPT in females and increased ALP & urea in males. The corresponding NOAEL is 100 ppm ( 25/ 37 mg/ kg/ day).

Treatment of SPF CF 1 / W 74 ® male and female mice with a dietary levels up to and including 500 ppm KWG 0599 for 24 months did not result in a significant increase in neoplasms compared to the control group.

This carcinogenicity study in the mice is Acceptable/ Guideline for a carcinogenicity study [ OPPTS 870.4200b; OECD 451] in mice. Although tests for homogeneity or stability and analyses of concentration of test material in the diet was not done such that dose levels can not be verified, the weight of evidence would suggest that this study can be classified as acceptable because ( 1) A battery of mutagenicity studies are all negative; ( 2) The HDT approached an adequate dose; ( 3) There was indications of increased incidence of liver adenomas, but not carcinomas at the top dose; ( 4) An acceptable Chronic Dog study showed a lower NOAEL; ( 5) The study supports a pesticide with limited exposure ( import tolerance on bananas only); and ( 6) The study is listed as acceptable TXR# 0002588. Additional deficiencies, which do not negate the study are listed in Section IIIC of the DER.

Table 4.4: Summary of Toxicological Doses and Endpoints for Chemical for Use in Human 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 ( Females 13 49 years of age) NOAEL = 50 mg/ kg/ day UF = 1000 Acute RfD = 0.05 mg/ kg/ day FQPA SF = 1X aPAD = acute RfD FQPA SF = 0.05 mg/ kg/ day Developmental Toxicity in rabbits MRID 40490801 Maternal; LOAEL = 150 mg/ kg/ day based on decreased body weight gain and food consumption. Developmental; LOAEL = 150 mg/ kg/ day based on increased potimplantation loss, decreased fetal weight, and reduced ossification.

Acute Dietary ( General population No endpoint from a single dose was seen in an appropriate study. Table 4.4: Summary of Toxicological Doses and Endpoints for Chemical for Use in Human Risk Assessments Exposure Scenario Dose Used in Risk Assessment, UF Special FQPA SF* and Level of Concern for Risk Assessment Study and Toxicological Effects Page 37 of 58 Chronic Dietary ( All populations) NOAEL= 2.11 mg/ kg/ day UF = 1000 Chronic RfD = 0.00211 mg/ kg/ day FQPA SF = 1X cPAD = chronic RfD FQPA SF = 0.00211 mg/ kg/ day Chronic/ dog [ MRID 00157466 & 00157465] LOAEL = 8.18 mg/ kg/ day based on based on adrenal vacuolation..

Incidental Short and Intermediate oral exposure There is no incidental oral short or intermediate exposure in US.

Short, Intermediate & Long term Dermal Endpoint not necessary for an import tolerance All Inhalation exposures No endpoint is necessary since there is no inhalation exposure in United States.

Cancer ( oral, dermal) " E," Unlikely human carcinogen. No dose related tumors in the rat or mouse.

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.

4.5 Special FQPA Safety Factor Because there is no residual uncertainty with respect to pre and post natal toxicity based on the submitted developmental and reproduction studies, the Special FQPA Factor is reduced to 1X. There is some uncertainty for potential neurotoxicity based on some literature studies. Because acute and subchronic neurotoxicity studies are not available, a 10X database uncertainty factor ( UF DB ) is retained to account for the lack of data.

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 Page 38 of 58 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 systems may be added to the Endocrine Disruptor Screening Program ( EDSP).

In the available toxicity studies with bitertanol, no evidence of agonistic or antagonistic estrogen or androgen or thyroid activity was seen. A potential hormonal antagonistic effects was seen as a prostate weight decrement in a subchronic dog study, but not in dog chronic studies. The change in prostate weight not seen at 1 year or 2 years in the chronic dog studies suggest that the effect in the 90 day dog study was specific to the dogs tested and not to bitertanol treatment. However, the reason for the difference in the 90 day and chronic dog studies is unknown.

When additional appropriate screening and/ or testing protocols being considered under the Agency's EDSP have been developed, bitertanol may be subjected to further screening and/ or testing to better characterize effects related to endocrine disruption.

5.0 Public Health Data 5.1 Incident Reports An incident report for bitertanol is not available.

6.0 Exposure Characterization/ Assessment S. Ary. Bitertanol: Summary of Analytical Chemistry and Residue Data for the Tolerance Reassessment Eligibility Decision ( TRED) Document. November 7, 2005. DP Barcode D302882.

6.1 Dietary Exposure/ Risk Pathway 6.1.1 Residue Profile The nature of residues in plants is adequately understood based on acceptable plant metabolism studies in apples and peanuts conducted with bitertanol labeled in the biphenyl ring system and additional studies submitted recently in apples, cotton, and tomatoes conducted with bitertanol labeled in the triazole ring system. The recently submitted studies are currently under review by the Agency; however, it is noted that after initial review of the newly submitted studies, the data support the conclusions drawn from the two previous submissions. The residues identified in these studies consisted primarily of the parent compound with traces of the metabolites bitertanol ketone and 4 hydroxybiphenyl. Although no metabolism studies for bananas are available, HED has concluded that the nature of residues in bananas is likely similar to that in apples, cotton, peanuts, and tomatoes. Therefore, HED has determined that the bitertanol residue of concern in/ on bananas for tolerance expression and for risk assessment is bitertanol ( parent only). Page 39 of 58 An analytical method for analysis of residues of bitertanol has been submitted and found acceptable for data collection purposes. The submitted method is a modification of Mobay Chemical Corporation's Method 54166 for the analysis of bitertanol in apples. Analysis is by gas chromatography using a nitrogen phosphorus flame ionization detector. The limit of detection is 0.01 ppm. A method trial was successfully performed on banana pulp and peels at fortification levels of 0.1 ppm and 0.2 ppm. Recoveries ranged from 78 to 88% for pulp samples, and from 70 to 98% for peel samples. The method was approved by the Agency for the enforcement of tolerance in/ on bananas and the results were forwarded to FDA for inclusion in the Pesticide Analytical Manual ( PAM) Volume II.

The FDA PESTDATA database dated 11/ 2001 ( PAM Volume I, Appendix I) indicates that bitertanol was completely recovered using Multiresidue Method Section 302 ( Protocol D).

Storage stability data for bitertanol in/ on bananas have not been submitted by the registrant; therefore, storage stability data are required.

As indicated in the tolerance petition, bitertanol was originally marketed for use on bananas in Costa Rica and Honduras, but has expanded to include Ecuador, Guatemala, Honduras, Mexico, Nicaragua, Peru, Venezuela, Dominican Republic, and Panama. Residue data from trials conducted in Costa Rica and Honduras have been submitted and reviewed by the Agency. Bitertanol was applied to both bagged and unbagged bunches of green and ripe bananas at a rate of 0.26 lb ai/ A/ application ( 2x the current rate per application), the maximum rate per application indicated on the product label at that time. Samples of peel, pulp and whole fruit were collected 3 days after the last of 9 treatments and 0 days after the last of 12 treatments. Washed and unwashed samples were analyzed for bitertanol by Method 54166, with modifications. The number of trials, the location of the trials within Costa Rica and Honduras, and the formulation type were not specified. No data were provided on how long the samples were stored prior to analysis. Residues found in samples of whole fruit from unbagged and bagged bunches of ripe and green bananas were all below the recommended tolerance level of 0.5 ppm. The residue data are deemed adequate to support a tolerance of 0.5 ppm in/ on bananas.

The SC formulation will be supported with additional field trials that are currently being conducted by the registrant. The studies are planned for completion in 2006.

Regulatory Recommendations and Residue Chemistry Deficiencies:

° Revise tolerance to 0.50 ppm to harmonize with Codex.

° Storage stability data are required to determine the stability of residues in bananas during cold storage. No data were provided on how long the banana samples from submitted field trial studies were stored prior to analysis.

° Field trial data for the SC formulation are required.

° Additional data are required in order for the submitted field trials conducted in Costa Rica and Honduras to be acceptable: the number of trials, location of the trials within the Page 40 of 58 countries, and the formulation type used A tolerance has been established under 40 CFR § 180.457 for residues of bitertanol in/ on bananas imported into the U. S. Field trials conducted in Costa Rica and Honduras showed residues below the existing tolerance level of 0.2 ppm. In order to harmonize with Codex, the established tolerance should be increased to 0.5 ppm.

Since the tolerance for bitertanol is an import tolerance, it should be listed in a separate subsection of 40 CFR indicating that there are no U. S. registrations for bitertanol. A summary of the bitertanol tolerance reassessment for bananas is presented in the following table:

Table 6.1( a): Tolerance Summary for Bitertanol.

Commodity Established/ Proposed Tolerance ( ppm) Recommended Tolerance ( ppm) Comments [ correct commodity definition]

Banana ( whole) 0.20 ( established) 0.5 Tolerance should be listed in a separate subsection of 40 CFR indicating that there are no U. S. registrations associated with bitertanol. [ Banana]

Codex/ International Harmonization The Codex Alimentarius Commission has established a maximum residue limit ( MRL) for bitertanol residues in/ on bananas at 0.5 mg/ kg. The Codex MRL for bitertanol is expressed in terms of residues of bitertanol only, as is the U. S. tolerance expression. Various enforcement methods are listed for bitertanol under the Codex system including PAM Multiresidue Method Section 302 ( Protocol D).

It is EPA's policy to harmonize its tolerances with the levels established by Codex provided that the Agency has sufficient information to make a determination that the Codex MRLs will be protective of the health of the U. S. public and meet FFDCA standards. FQPA requires EPA to publish a notice for public comment whenever the Agency establishes a tolerance that differs from an established Codex MRL. The established tolerance should be increased to 0.50 ppm to harmonize with Codex as the dietary exposures and risks are not of concern ( S. Ary, D302880, 11/ 7/ 2005).

6.1.2 Acute and Chronic Dietary Exposure and Risk S. Ary. Acute and Chronic Dietary Exposure Assessments for the Tolerance Reassessment Eligibility Decision ( TRED) Document. November 7, 2005. DP Barcode D302880.

Acute dietary risk estimates are provided for females 13 49 years of age only. For the general population there is no study to support an effect from a single dose. The acute dietary risk assessment concludes that for the supported use of bitertanol on imported bananas, the acute Page 41 of 58 dietary risk estimate does not exceed HED's level of concern ( i. e., dietary risk is less than 100% of the aPAD) at the 95th exposure percentile for females 13 49 years old ( 2% of the aPAD).

Chronic dietary risk estimates do not exceed HED's level of concern ( i. e., dietary risks are less than 100% of the cPAD) for the U. S. population ( 9% of the cPAD) and all population subgroups. The highest exposed population subgroup is children 1 2 years of age, at 43% of the cPAD.

The results of the acute and chronic dietary exposure analysis are presented in the following table.

Table 6.1( b): Summary of Acute and Chronic Dietary Exposure and Risk for Bitertanol.

Population Subgroup a Acute Dietary ( 95th Percentile) Chronic Dietary Cancer Dietary aPAD, mg/ kg Exposure, mg/ kg/ da y % aPAD cPAD, mg/ kg/ da y Exposure, mg/ kg/ day % cPAD Exposure mg/ kg/ da y Risk General U. S. Population Not Applicable 0.00211 0.000179 9 Not Applicable All Infants (< 1 yr) 0.00211 0.000731 35 Children 1 2 yrs 0.00211 0.000917 43 Children 3 5 yrs 0.00211 0.000467 22 Children 6 12 yrs 0.00211 0.000192 9 Youth 13 19 yrs 0.00211 0.000064 3 Adults 20 49 yrs 0.00211 0.000104 5 Adults 50+ yrs 0.00211 0.000177 8 Females 13 49 yrs 0.05 0.000964 2 0.00211 0.000105 5 a The values for the population with the highest risk for each type of risk assessment are bolded.

6.2 Water Exposure/ Risk Pathway Bitertanol has no registered uses in the United States; therefore, a drinking water assessment has not been conducted.

6.3 Residential ( Non Occupational) Exposure/ Risk Pathway Bitertanol has no registered uses in the United States; therefore, a residential exposure assessment has not been conducted.