Buyer CropSeienee

NR 481097-01

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Test Order/DCI# EDSP-109901 -156

if)

Title:

150-DAY RESPONSE TO ERA'S TEST ORDER FOR

ENDOCRINE SCREENING ON TRIADIMEFON

Guideline:

OPPTS 890 Guideline Series

Authors:

Ann M. Blacker, PhD, DABT

Tilghman Hall, PhD

Patricia Kwiatkowski, PhD

Date:

May 25, 2010

Report ID:

US0046

Bayer CropScience LP

P.O. Box12014

Research Triangle Park, NC 27709

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STATEMENT OF NO DATA CONFIDENTIALITY

No claim of confidentiality is made for any information contained in this study on the basis of its

falling within the scope of FIFRA 10(d)(l)(A), (B) or (C).

Submission of these data in compliance with FIFRA does not constitute a waiver of any right to

confidentiality which may exist under any other statute in any country other than the USA.

The above statement supersedes all other statements of confidentiality that may occur elsewhere

in this report.

Company: Bayer Environmental Science

A Division of Bayer CropScience LP

Company Agent: Date:

Richard M. Gorrell, Ph.D.

Registration Product Manager
< NJttSrllMU I*

US0046

GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT

This report is not subject to the principles of 40 CFR 160, Good Laboratory Practices

Standards (FIFRA), as promulgated in Federal Register, final rule. August 17.1989.

5/

Ann M. Blacker. PhD, DABT Date

Toxicdogist

Bayer CropScience

TilghmartWall, PhD Date

Ecotoxicologist

Bayer CropScience

Patricia Kwiatl

Product Safety

Bayer CropScience

Richard M. Gorrell Date

Manager, Registrations

Bayer Environmental Science

A Division of Bayer CropScience LP

Sponsor/Submitter

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150-DAY RESPONSE TO ERA'S TEST ORDER FOR

ENDOCRINE SCREENING ON TRIADIMEFON

Introduction:

On December 17, 2009, EPA issued a Test Order for endocrine screening on triadimefon. In

this Order, EPA provides the option to cite or submit existing data to satisfy part or all of the

testing. Specifically, EPA states,"... such hazard-related data would be of high quality and

achieves the objective of Tier 1 assays to provide reasonable assurance that a chemical does

or does not have the potential to interact with the estrogen, androgen, or thyroid systems." The

Triadimefon Consortium was established to address EPA's data call-in and includes the

following manufacturers: Bayer CropScience and Tide International.

Studies conducted for regulatory purposes as well as studies published in the open literature

were identified and reviewed by Bayer CropScience on behalf of The Consortium. An objective

assessment of these studies was performed to determine whether, as a body of data, they

provide data functionally equivalent to that in EPA's EDSP Tier 1 screening battery and include

sufficient information to characterize potential endocrine-modulating effects on estrogen,

androgen or thyroid hormone pathways. When evaluating information reported in the literature,

in general, the approach of Klimisch et al. (1997) has been utilized to systematically assess the

quality of work reported. This approach is recognized and used by regulatory authorities

including its use in Europe in developing the International Uniform Chemical Information

Database (IUCLID). The data reliability criteria presented by Klimisch et al. are similar to the

criteria recommended by EPA in the High Production Volume Program under TSCA. Assays

reported in the literature have also been compared to the relevant assay described in the

OPPTS 890 Guideline Series to identify major differences between the specific assay included

in EPA's validation program for Tier 1 screening and the one reported in the literature.

The Triadimefon Consortium is citing multiple studies and publications as "Other Scientifically

Relevant Information" or OSRI, which provide data that are functionally equivalent to, and

effectively substitutes for, the information that would be developed through many of the EDSP

Tier 1 screening assays. We believe that the specified data, in conjunction with conduct of two

Tier 1 assays, will provide a test battery that conserves animal usage, uses existing information

appropriately, and will adequately identify any potential endocrine modulating effects of

triadimefon. Much of the existing data comes from US EPA published research studies and

guideline or guideline equivalent mammalian toxicology regulatory studies which have been

reviewed by EPA and have been classified as acceptable in the Triadimefon RED and

subsequent risk assessment documents. Endocrine-related apical endpoints which have been

assessed in the guideline studies on triadimefon that are functionally equivalent to endpoints

assessed in EPA's Tier 1 endocrine screening battery are summarized in Table 1a & b.

Functionally equivalent, in this context, refers to data of a suitable nature and quality to provide

the same essential predictive information on a chemical even if different methods and

procedures may be used for obtaining the data. Further, the majority of the endpoints required

in the pubertal assays have been assessed, taking into account both existing guideline studies

and relevant findings recently reported by EPA research staff in the open literature as shown in

Table 2 and Table 3. Additionally, there is a significant body of data on thyroid function and

histopathology, much of it from EPA research publications, to serve as an adequate surrogate

for the amphibian metamorphosis assay as shown in Table 4.

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The Triadimefon Consortium believes that existing data, as reported in both guideline studies

and research publications, are for the most part adequate to assess the potential for this

compound to interact with estrogen, androgen and thyroid systems. The Triadimefon

Consortium believes that conducting the following two assays from the EDSP Tier 1 screening

battery will provide complementary data and sufficient scientific redundancy to assure that

potential adverse endocrine effects would be identified and sufficient data are available to

perform a weight of evidence (WOE) evaluation of whether any Tier 2 testing would be

necessary:

In vivo assays: Fish short-term reproduction test

Hershberger bioassay

The principle of the fish short-term reproduction screen is to measure the reproductive

performance of fathead minnows as the primary indicator for endocrine disruption (EPA 2009

890.1350). The assay screens for disturbances in the hypothalamic-pituitary-gonadal (HPG)

axis including (anti-) estrogenic, (anti-) androgenic, aromatase inhibition, steroid modulation

effects, examining abnormalities associated with survival, reproductive behavior, secondary sex

characteristics, histopathology, and fecundity (i.e., number of spawns, number of eggs/spawn,

fertility, and development of offspring) offish exposed to test chemicals (USEPA Order/DCI

#EDSP-041101-42). Equivalent endpoints are not available with in vivo organisms in either

existing guideline ecotoxicological studies for triadimefon or in research studies reported in the

public domain, and therefore the Consortium will conduct the fish short-term reproduction assay.

The Hershberger assay is a short-term in vivo assay to detect androgenic or antiandrogenic

chemicals or chemicals that inhibit 5a-reductase by measuring weight changes in five

androgen dependent tissues: ventral prostate (VP), seminal vesicle (SV) (plus fluids and

coagulating glands), levator ani-bulbocavernosus (LABC) muscle, paired Cowper's glands

(COW) and the glans penis (GP). Sufficient data on these specific tissues are not available

in guideline or in research studies reported in the public domain, and therefore the Consortium

will conduct the Hershberger assay.

Following are assessments of the existing OSRI and The Triadimefon Consortium's position on

their sufficiency to meet the requirements of the various endocrine screening assays other than

the fish short-term reproduction assay and Hershberger assay, beginning with a summary of the

guideline studies being cited.

Summary of Triadimefon Guideline Studies

All studies summarized below have been reviewed by US EPA and judged acceptable

(Triadimefon. Preliminary Human Health Risk Assessment (Revised), DP Barcode D326678.

February 9, 2006 and Triadimefon: Human Health Risk Assessment to Support Reinstatement

of Use on Residential Turf, Decision No. 391164. July 24, 2009).

Relevant data which are functionally equivalent to address the purpose of the various endocrine

related assays in EPA's Tier 1 screening battery are available from guideline studies including

two rat reproduction studies, a developmental neurotoxity study, developmental toxicity studies,

subchronic toxicity studies in the rat and dog, lifetime feeding studies in rats and mice and a 2

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year chronic dog toxicity study. Functionally equivalent, in this context, refers to data of a

suitable nature and quality to provide the same essential predictive information on a chemical

even if different methods and procedures may be used for obtaining the data.

In the first multigeneration reproductive toxicity study with triadimefon (Loser, E., Lorke, D.,

1979; MRID 32541) male and female rats were administered triadimefon continuously in the diet

at concentrations of 0, 50, 300 or 1800 ppm and the reproductive performance studied over 3

consecutive generations, each with 2 matings. Offspring of each first mating (F1a, 2a and 3a)

were sacrificed on PND 28; offspring from the second matings were used for subsequent

matings (F1b, 2b received treated diet from weaning until approximately PND 100 and then

were mated, with females continuing to receive treated diet through gestation and lactation) or

offspring histopathology investigation (F3b). In a complementary two-generation reproductive

toxicity study (Eiben, R., 1984; MRID 155075, 92188019 & 92188320), triadimefon was

administered continuously in the diet to rats in the diet at levels of 0, 50 or 1800 ppm as follows:

P animals were given test article diet formulations from 100 days prior to mating to produce the

F1 generation. After weaning, F1 animals were selected to become the F1 parents of the F2

generation and were given the same concentration test formulation following the same pattern

as their dams except that only the 50 ppm F1 parents were sacrificed when the F2 litters were

weaned. Treatment of the F1 high dose groups was stopped and the high dose F1 males were

mated with the control F1 females, while the high dose F1 females were mated with control F1

males. This cross-mating procedure produced a second F2 litter and was intended to provide

further information regarding the fertility of the F1 parents.

For all matings, the day that sperm was detected in a vaginal smear was designated as

gestation day (GD) 0. Sperm motility, sperm morphology, gestation length, and sex ratios were

unaffected by treatment.

When administered at levels up to 300 ppm, triadimefon had no effect on mating, fertility,

pregnancy or lactation performance of PO, F1 or F2 females. There were no adverse effects on

pup weights at birth or survival rates during lacation in F1, F2 or F3 pups. In the first study, the

dietary level of 300 ppm had slight effects on F2b and F3b pup body weights during lactation.

No histopathological changes were observed in any organ examined (eyes, adrenals, brain,

thyroid, thymus, heart, lung, liver, spleen, pancreas, lymph nodes, stomach, intestine, kidneys,

testes, epididymis, ovaries, uterus) from four week old F3b pups which were euthanized and

necropsied.

In both studies, at 1800 ppm fertility was affected in all generations. Body weights of P, F1 and

F2 generation parents were decreased during premating, mating, gestation, and lactation. Body

weight gains of the F1 generation parental males and females were also decreased throughout

pre-mating, gestation, and lactation for both matings. In the first study, the number of offspring

per dam at birth was decreased in the first litters of both the P generation and the F1

generation. Fertility values were significantly lower in both matings of the F1 generation. Only

one female became pregnant in the F2a mating, and no females became pregnant in the F2b

mating. In the second study, at 1800 ppm, fertility was also significantly decreased in the F1

parents. Additionally, body weights were decreased in the F1 parental males and females

during weeks 9-40 and body weights were decreased at birth and during lactation in the F1 and

F2 pups. F2 litter size was decreased; viability was decreased in the F1 and F2 pups during

lactation. Elevated serum testosterone levels were reported for high dose F1 males at necropsy,

but luteinizing hormone and estradiol levels were unaffected. No histopathological changes

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were observed in any reproductive organs examined (testes, ovaries, epididymis, pituitary,

prostate, seminal vesicles, uterus and vagina) from necropsied F1 parents or F2 pups.

In the F1 cross-mating (second study), fertility was 47.4% from the mating of high dose males

with the control females and 80% from the mating of the high dose females with the control

males. In the absence of sperm abnormalities, it was concluded that the observed decrease in

fertility was due to a decreased interest of the males in copulation (copulation was not observed

for 6/10 of the 1800 ppm males during the 3-week cross-mating with control females).

In a developmental neurotoxicity study with triadimefon (Sheets, L, et al., 2008; MRID

47377101), pregnant rats were administered triadimefon in the diet at levels of 0, 100, 300 or

800 ppm from GD 6 to lactation day (LD) 21 when pups were weaned and put onto untreated

diet. Offspring were evaluated for a variety of parameters including body weight, acquisition of

vaginal patency and preputial separation, surface righting, pupil constriction, acoustic startle,

motor activity and learning and memory. Neural tissues were taken at post natal day (PND) 21

and 75 for microscopic examination and morphometry.

Parental reproduction parameters, litter size and viability were unaffected by treatment. The

acquisition of vaginal patency and preputial separation in F1 offspring was unaffected at

maternal dietary levels up to 800 ppm, the highest dose tested; offspring surface righting and

pupil constriction were also unaffected. Motor and locomotor activity was unaffected for both

sexes in all dose groups. At 800 ppm, there were decreases in pre-weaning body weight (PND

11, 17, and 21) in both sexes, and pre-weaning body weight gain in both sexes; deviated snout

was seen in 3 females and 1 male. An increased peak amplitude during auditory startle reflex

testing was observed in high dose females only on PND 60 and increased number of trials to

criterion during the retention phase of the avoidance test in females as well. There were no

differences in micropathology brain measurements in day 21 or 75 animals considered to be

treatment related.

In developmental toxicity studies investigating the effect of triadimefon on the developing

fetus in rats, there were ne effects on the number of corpora lutea/dam, implants/dam, viable

fetuses, early or late res«rptions, non-viable implantations/dam, live litters, fetuses/dam or on

sex ratio at maternal d»ses up to 90 mg/kg bw/day in one study (Linger, T. et al.,1982; MRIDs

149336 & 92188018) and 100 mg/kg bw/day in the other (Nagumo, K. et al., 1981; MRID

89023) even in the presence of significant maternal toxicity as evidenced by decreased body

weight gains and increased motor activity. In a rabbit developmental toxicity study (Clemens, G.

and Hartnagel, R., 1990; MRID 41446201 & 42089601) with triadimefon there was also no

treatment-related effect on gestation or fertility indices at maternal doses up to 120 mg/kg

bw/day.

There were no overt signs of toxicity in subchronic rat (Mohr, U., 1976; MRID 48624 ) and

subchronic dog (Hoffman, K. and Luckhaus, G., 1974; MRID 48625 & 60226) toxicity studies

at the highest doses tested (100 and 60 mg/kg bw/day, respectively). There were no treatment

related effects noted on adrenals, thyroid, pituitary, ovary or testis tissues examined at the

highest doses tested.

In 21 month mouse oncogenicity (Bomhard, E., 1986; MRID 40752101 & 40865101 ), 24

month rat toxicity/oncogenicity (Bomhard, E. and Schilde, B., 1991; MRID 42153901) and 24

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month dog toxicity (Hoffman, K. and Groning, P., 1978; MRID 00032539 & 00126261) studies,

there were no treatment related effects noted on any reproductive organ examined at any dose

level (highest doses tested: mouse, 765 and 550 mg/kg bw/day for females and males,

respectively; rat, 199 and 114 mg/kg bw/day for females and males, respectively; dog, 68.8 and

60.4 mg/kg bw/day for females and males, respectively).

In the mouse oncogenicity study, the LOAEL is 119/76 mg/kg bw/day (f/m), the middle dose

tested, based on liver hypertrophy in both sexes and other sex specific liver anomolies, with a

treatment related increase in heptacellular adenomas. In the rat combined chronic

toxicity/oncogencity study, the LOAEL is equivalent to 199/114 mg/kg bw/day (f/m), the highest

dose tested, based on: increased food consumption and incidences of fat in hepatocytes in both

sexes; increased alanine aminotransferase levels in males; increased absolute and relative liver

weight and cholesterol levels in females and decreased body weight and body weight gains in

females. The carcinogenic potential of triadimefon in rats was considered equivocal by US EPA

based on the incidence of thyroid adenomas and cystic hyperplasia in both sexes, which were

suggested by BCS to be within historical control range (Sander, E. and Schilde, B., 1993; MRID

43089301). For the 2 year dog study, the LOAEL is 34.70/33.67 mg/kg bw/day( m/f), the middle

dose tested, based on increased alkaline phosphatase levels in both sexes; and decreased food

consumption and increased cholesterol in females.

The Cancer Assessment Review Committee (CARC) classified triadimefon into category C

"possible human carcinogen" - based on a statistically significant increase in thyroid adenomas

in male Wistar rats and statistically significant increases in hepatocellular adenomas in

both sexes of the NMRI mouse. However, it was concluded that a quantified carcinogenic risk

assessment for triadimefon is not appropriate and risk assessment will be based on the cPAD

and Margin of Exposure approach only.

Other Scientifically Relevant Information

Estrogen System OSRI

»

Mammalian assays (estrogen receptor (ER) binding, ER transcriptional activation, uterotrophic,

pubertal female) have been included in the EDSP Tier 1 battery to assess the potential of a

chemical to affect the estrogen signaling pathway. None of the assays as specifically described

in EPA's OPPTS 890 Guideline Series have been conducted with triadimefon. However, as

shown in Tables 1a & 1 b, data relevant to ER binding and transcriptional activation,

uterotrophic, and pubertal female assays are available from guideline studies, all of which have

been reviewed by EPA and classified as acceptable. Estrogenic and anti-estrogenic activities

have been examined independently by two research groups in an MCF-7 cell proliferation assay

(Vinggaard et al., 1999; Okubo et al., 2004). In addition, triadimefon was included in a Japanese

investigation that screened 200 pesticide chemicals for estrogen and androgen activities

(Kojima et al., 2004). The three publications relevant to ER activity are detailed below. Also,

data from the two regulatory multi-generation rat reproduction studies and relevant results from

a recent publication authored by US EPA scientists reporting effects of conazole fungicides on

reproductive development in female rats (Rockett et al., 2006) are presented to adequately

address the requirement for a female pubertal assay (Table 2).

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In vitro Estrogen Receptor Binding Assay and Estrogen Receptor Transactivation Assay

Although no studies have been conducted according to the 890 Guidelines issued by EPA,

specifically OPPTS 890.1250 and 890.1300, estrogenic and anti-estrogenic activities have been

examined independently by two research groups in an MCF-7 cell proliferation assay

(Vinggaard et al., 1999; Okubo et al., 2004). A recombinant yeast strain (Saccharomyces

cerevisaie), stably transfected with the human estrogen receptor a and a reporter plasmid

encoding for the enzyme (3-galactosidase (originally developed by Glaxo-Wellcome) was also

used by Vinggaard to examine triadimefon's estrogenic activity. Although triadimefon did not

produce any response in the yeast screen, the lack of activity was attributed to cytotoxicity.

In both cell proliferation experiments, human breast cancer estrogen-sensitive MCF-7 cells were

exposed to a series of triadimefon concentrations (Vinggaard: 0.001, 0.1,1 and 10uM,

dissolved in medium; Okubo: 6 concentrations ranging from 1 to 1000uM, dissolved in ethanol:

combining both studies, a total of 8 concentrations were tested ranging from 10~9 to 10~3 M). In

both experiments, 17 (3-estradiol was used as the positive control and reference to calculate

relative proliferation potency (RPP) and relative proliferation efficiency/effect (RPE). The Okubo

investigation included cytotoxicity information; triadimefon was reported not to be cytotoxic at

concentrations up to 1000uM. In both experiments, triadimefon induced an increase cell

proliferation and similar RPEs were reported. Both publications identified triadimefon as a weak

estrogen receptor agonist.

Additional experiments were conducted and reported in the Okuba et al. publication. The effect

of antiestrogen ICI 182,780 on cell proliferation induced by triadimefon was also examined as

well as the antiestrogenic activity of triadimefon as determined by the suppression of cell

proliferation induced by 30 pM 17 (3-estradiol. Addition of agonist ICI 182,780 significantly

decreased the cell-proliferative effect of triadimefon to approximately the control (0.1% ethanol)

level. In the antiestrogen assay, triadimefon was antiestrogenic at 100 and 1000 uM. Finally, the

binding affinity of triadimefon for human ERa or AR was determined with ligand screening

receptor systems. There was so little binding of triadimefon to ERa that an IC50 value could not

be calculated, and the IC50 value of triadimefon to AR was 902 ± 143 uM with an RBA (relative

binding affinity compared to positive control mibolerone) of 0.005 (IC50 values being the

concentration of compound that inhibits binding of 17 (3-estradiol to ER or that of testosterone to

AR by 50%).

Results of another ER assay with triadimefon have been reported in the literature. Kojima et. al.

(2004) investigated the potential for 200 pesticides, including triadimefon, to bind to the alpha

and beta ER using transfected Chinese Hamster Ovary-K1 cells. Triadimefon was tested at

concentrations ranging from ~10~8 to 10"5 M, and the REC20 (concentration of chemical that

results in 20% of the agonist activity of 10"10 M E2) value and relative estrogenic activity at

10~5 M concentration (RLA) were measured. Triadimefon was not reported to show estrogenic

activity. However, it must be noted that the authors tested fewer than the eight concentrations

required by the OPPTS 890.1250 Guideline and did not provide information on solubility or

cytotoxicity. Thus, these results cannot be considered reliable on their own, but are consistent

with the conclusion that estrogen receptor activation is not an important mode of action for

triadimefon.

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Another study examined the interaction of triadimefon with the rainbow trout estrogen receptor,

using a recombinant yeast cell line, as well as rainbow trout hepatocyte aggregate cultures

(Petit at al. 1997). Test concetrations for triadimefon (and 49 other indepently tested

compounds) ranged from 10~8 to 10"4 M. The yeast cell line showed similar results as above

with no activity with the receptor, most likely to due to cell death. With the trout hepatocyte cell

line, the study does not provide the results of all tested concentrations, and therefore

interpretation of the results in limited. However, it appears that triadimefon did not displace

estradiol binding to the trout estrogen receptor, however vitellogenin induction was noted. The

authors suggest that that activity (vitellogenin induction) was due to either a metabolite or to an

indirect gene activiation pathway.

Based upon the consistency of the findings in three independently conducted in vitro ER assay

experiments, the requirement for the EDSP Tier 1 in vitro estrogen receptor binding assay and

estrogen receptor transactivation assay should be waived.

In vivo Female Pubertal Assay:

The effects of triadimefon on the reproductive development in the female Wistar Han rat were

also investigated by US EPA researchers (Rockett et al., 2006). Mated female rats were

administered triadimefon via the diet (100, 500, or 1800 ppm) from gestation day 6 through

lactation. Female offspring were exposed via lactation until PND 22 and then received the same

dietary concentrations as their dams until PND 98. Thus, the exposure duration for the F1

females exceeds that required by the pubertal assay and therefore provides a more severe

assessment of a compound's potential to affect endocrine-related endpoints. Body weight (BW)

and anogenital distance (AGO) at PND 0, age and BW at vaginal opening (VO), estrous

cyclicity, and body and various organ weights at necropsy were said to be measured. Liver,

ovaries and thyroid were examined microscopically.

Triadimefon administered at 500 ppm in the diet had no significant adverse effect on dams or

offspring. Offspring anogenital distance on day of birth was unaffected at all treatment levels;

VO was comparable to controls for low and mid-dose females (consistent with the findings in the

triadimefon developmental neurotoxicity study (Sheets, L, 2008) where no effect on VO in

females or pre-putial separation (PPS) in males was noted in pups born of mothers receiving

triadimefon levels at up to 800 ppm in the diet from GD6 to LD21). The reported delay in VO in

high dose females is confounded by the absence of body weight data in the publication. The

authors state that body weights at birth and VO were unaffected by triadimefon but at necropsy

(PND 99) weights of high dose animals were significantly lower than the controls. It seems

improbable that the developing pup body weights for the high dose group were unaffected by

treatment, given both the statistically significant decrease in food consumption from weaning to

necropsy as reported in the publication detailed in Table 3 (in particular from PND 29-43,

p<0.001) in addition to the reported statistically significant reduced body weights

(p<0.001PND1, 8 and <0.0001 PND 22 to 92) reported for high dose male pups in the male

cohort study (Goetz et.al. 2007), which is summarized later. In addition, as presented earlier, in

both regulatory multi-generation reproduction studies, adult rats receiving 1800 ppm of

triadimefon in their diet as well as offspring had significantly reduced body weights over most if

not all of their life spans. Therefore the significance of the reported VO delay in high dose

offspring is questionable in the absence of body weight data. Further, in the Rockett publication,

relative liver and ovary weights were reported increased in female offspring at the high dose.

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Only mean relative organ weights are mentioned in the report when discussing significant

differences between high dose animals and control; absolute organ weights and body weights

are needed to understand the toxicological significance of changes in relative organ weights.

Histopathology showed liver and thyroid effects considered to be treatment related. There were

no unusual observations for ovaries; all ovaries contained follicles at all stages of development,

had normal appearing corpora lutea and showed no signs of edema, cystic follicles or retained

oocytes.

In the two multi-generation rat studies (Loser and Lorke, 1979: Eiben, 1984) P adults were

exposed to triadimefon in the diet for at least 10 weeks prior to mating and then throughout

mating, gestation, and lactation periods. Indirect exposure for the F1 offspring covers the in

utero and lactation periods with direct exposure starting around PND 11 or 12 when pups begin

to consume treated diet and from PND 28, continues for at least 10 weeks premating throughout

mating, gestation, and lactation periods for pups selected to become F1 parents. Thus, the

exposure duration for the F1 generation exceeds that required by the pubertal assay and

therefore provides a more severe assessment of a compound's potential to affect endocrinerelated endpoints. The results of these studies have been discussed above. There were no

fertility or sex organ related findings in either males or females that suggest estrogen disruption.

The pubertal assay is designed to detect effects on pubertal development and thyroid function.

As detailed in Table 2, all of the sexual developmental endpoints included in a female pubertal

assay have been assessed for triadimefon. With the exception of clinical chemistry and thyroid

hormone measurements, the endpoints in the female pubertal assay are captured in the "higher

tiered" two-generation rat reproduction studies and research results presented by Rockett et al.

(2006). Complete clinical chemistry data are available for multiple timepoints in the chronic rat

and dog studies. Thyroid weight and histology are available from chronic studies in rats, mice

and dogs as discussed earlier. Thyroid hormone level measurements are not available for

female rats but have been measured in male rats by EPA researchers and will be discussed

later in relation to the androgen assays. The results from all of these studies, in particular the

two-generation rat reproduction studies, developmental neurotoxicity study and 2006 publication

by Rockett, et al., should be considered adequate to address the data requirement for a female

pubertal assay. Therefore, the need to conduct this assay should be waived.

In vivo Uterotrophic Assay

EPA states that uterotrophic response is not due exclusively to estrogenic chemicals so that a

Uterotrophic response should be confirmed by corroborating information such as ER binding or

transcriptional activation. Findings in published transcriptional activation studies did not suggest

that estrogen receptor activation is an important mode of action for triadimefon. In addition,

neither the multi-generation reproduction studies nor the research by Rockett et al. suggest

estrogenic activity. Therefore, the need to conduct this assay should be waived.

Conclusion for ER Assays

Adverse effects that would be expected from exposure to an estrogen disrupting chemical are

shown below. None of the effects are consistent with what has been observed in mammalian

toxicology studies with triadimefon.

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Mechanism Potential Effects in Males Potential Effects in Females

Estrogenicity Delayed preputial separation

(marked, or in absence of significant

body weight decreases)

Reduced fertility

Reduced sperm counts

Decreased male reproductive organ

weights (except prostate weight,

which may be increased)

Histopathological findings of the

testes (e.g., Leydig cell proliferation

and/or tumors)

Increased incidence, growth of

pituitary tumors

Precocious vaginal opening

Persistent estrus

Increased time to mating

Reduced gestation duration

Increased uterine weights

Histopathological findings of the

female reproductive organs (e.g.,

vaginal cornification, uterine

hypertrophy and hyperplasia)

Increased mammary tumors

(adenomas or adenocarcinomas)

Increased incidence, growth of

pituitary tumors

Ovarian, uterine and vaginal tumors

in female offspring

Anti-estrogenicity Decreased epithelium height in

testicular tubules

Testicular weight increases

(short term)

Testicular atrophy

(from long-term exposure)

Infertility and testicular atrophy

(from moderate-term exposures)

Delayed vaginal opening

Delayed start of estrous cycling;

irregular or absent estrous cyclicity

Reduced fertility

Decreased corpora lutea,

implantations

Decreased female reproductive

organ weights

Decreased mammary tumor

incidence

In summary, scientific information indicates that an estrogenic or anti-estrogenic compound

would produce among other findings disturbances in male fertility/sperm count or motility,

estrous cyclicity and/or alterations in the weight and histopathology of specific reproductive

organs (e.g., ovary, uterus, testis, or epididymis). Adequate information on triadimefon effects

on all parameters is available in existing regulatory studies and publications authored by EPA

researchers. Given the level of available information, it is considered unnecessary to conduct

either of the in vitro estrogen system assays or to sacrifice the additional animals needed to

conduct the female pubertal assay or the uterotrophic screening assay. Therefore, the

requirement for the estrogen-based EDSP Tier 1 mammalian assays should be waived.

While the rainbow trout hepatocyte cell lines may have demonstrated an induction of

vitellogenin (Petit et al. 1997), the mechanism for such induction was not clear. For this reason,

the Triadimefon Consortium has agreed to conduct the fish short-term reproduction test.

Androgen System OSRI

Mammalian assays (androgen receptor (AR) binding, Hershberger, pubertal male) have been

included in the EDSP Tier 1 battery to assess the potential of a chemical to affect the androgen

signaling pathway. None of the assays as specifically described in EPA's OPPTS 890 Guideline

Series have been conducted with triadimefon. However, as shown in Tables 1a & 1 b, data

relevant to AR binding, Hershberger and pubertal male assays are available from guideline

studies, all of which have been reviewed by EPA and classified as acceptable. Androgenic

activity has been examined with a ligand screening androgen receptor system by Okubo et al.,

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2004 as reported above. In addition, triadimefon was included in a Japanese investigation that

screened 200 pesticide chemicals for estrogen and androgen activities (Kojima et al., 2004).

Also, data from the two regulatory multi-generation rat reproduction studies and relevant results

from a recent publication authored by US EPA scientists reporting on the reproductive toxicity of

conazole fungicides in male rats (Goetz et al., 2007) are presented to adequately address the

requirement for a male pubertal assay (Table 3).

In vitro Androgen Binding Assay

As reported above, both Okuba et al., 2006 and Kojima et al., 2004 included triadimefon in

androgen screening assays. Although neither was conducted according to the EPA protocol for

the Tier 1 assay, the fact that both assays showed essentially no binding should be sufficient

evidence that triadimefon does not act at the androgen receptor.

In vivo Male Pubertal Assay

The reproductive toxicity of triadimefon on in the male Wistar Han rat was investigated by US

EPA researchers (Goetz et al., 2007). Mated female rats were administered triadimefon via the

diet (100, 500, or 1800 ppm) from gestation day 6 through lactation. Male offspring were

exposed via lactation until PND 22 and then received the same dietary concentrations as their

dams until PND 92. Thus, the exposure duration for the F1 males exceeds that required by the

pubertal assay and therefore provides a more severe assessment of a compound's potential to

affect endocrine-related endpoints. BW and AGO were measured on PNDO. Food consumption

and BW were measured weekly. Age and BW were recorded on the day of PPS. One male from

each litter was taken to necropsy at PND1, 22, 50 and 92 to assess effects on organ weights

(brain, hypothalamus, pituitary, thyroid, liver, testis, epididymis, ventral prostrates and seminal

vesicles), histology (brain, pituitary, thyroid, liver, testis, epididymis and ventral prostrates) and

hormone measurements (estradiol, testosterone, total triiodothyronine (T3), total thyroxine (T4),

thyroid stimulating hormone (TSH) and luteinizing hormone (LH). Due to the limited numbers of

high dose males surviving to weaning, a PND50 necropsy was not done for the high dose

group. One testis and one epididymis from each male at PND22, 50 and 92 necropsy were used

for sperm morphology and motility analysis. Mating assays were conducted on PND78 or older

treated males with untreated females. Ejaculated sperm counts after natural breeding were

measured for high dose males.

There were no treatment related effects on growth or survival at 500 ppm and below. High dose

male offspring had reduced body weight from birth and remained less than control throughout

the study (statistically significant at all times). Survival rate in the high dose group was reduced

to 47%; there were no deaths in animals that survived past PND8. AGO was significantly

increased in males in the 1800 ppm group, and PPS was delayed, a finding confounded by

reduced pup size/BW. There were no effects noted in mid- and low-dose pups.

Sperm head and tail morphology and sperm motility were unaffected by treatment at all levels.

As observed in the multigeneration studies, males receiving 1800 ppm apparently did not mate;

no effect on insemination index or percent fertility was noted for males receiving 100 or 500

ppm, with all fertility parameters in the pregnant females comparable to controls. Elevated

serum testosterone levels were reported for high and mid- males at necropsy, but luteinizing

hormone and estradiol levels were unaffected, again consistent with multi-generation study

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findings. T4 levels in 1800 ppm males were decreased at PND92 only but neither T3 nor TSH

levels were affected. Relative, but not absolute, liver weights were increased in PND1 offspring

at all dose levels, in mid-dose males at PND50 and only in high dose males at PND92. Absolute

and relative increases in testes weights were observed in 500 ppm males at PND22 and 50 and

relative weights only were increased in 1800 ppm males at PND92. Relative prostrate weight

was increased in 100 and 500 ppm groups on day 92; absolute epididymal and seminal vesicle

weights were decreased for 1800 ppm males at PND92. Pituitary, thyroid, testis, epididymis and

prostate histology results showed no alterations across the treatment groups; liver histology

showed high dose males with strong panlobular hypertrophy at PND92 (4/4 animals), which is

consistent with findings in the second multigeneration reproduction study.

In two multi-generation rat studies (Loser and Lorke, 1979: Eiben, 1984), P adults were exposed

to triadimefon in the diet for at least 10 weeks prior to mating and then throughout mating,

gestation, and lactation periods. Indirect exposure for the F1 offspring covers the in utero and

lactation periods with direct exposure starting around PND 11 or 12 when pups begin to

consume treated diet and from PND 28, continues for at least 10 weeks premating throughout

mating, gestation, and lactation periods for pups selected to become F1 parents. Thus, the

exposure duration for the F1 generation exceeds that required by the pubertal assay and

therefore provides a more severe assessment of a compound's potential to affect endocrinerelated endpoints. The results of these studies have been discussed above. There were no

patterns of fertility or sex organ related findings in either males or females that suggest

androgen disruption.

The pubertal assay is designed to detect effects on pubertal development and thyroid function.

As detailed in Table 3, all of the sexual developmental endpoints included in a male pubertal

assay have been assessed for triadimefon. The results of these studies have been discussed

above. There was no obvious pattern of fertility or sex organ related findings in either males or

females that indicate disruption of the androgen system.

With the exception of weights of accessory tissues and clinical chemistry, the endpoints in the

male pubertal assay are captured in the two-generation rat reproduction studies and the 2007

published research by Goetz et al.. In addition, complete sperm analyses was performed in both

regulatory and research studies detailed above as well as in a study published by Tully, et al. in

2006 in which adult male SD rats (6 per dose group) received triadimefon by gavage from PND

60 to 74 at dose levels of 5, 50 or 115 mg/kg bw/day without adverse impact on sperm

morphology or motility, testis or epididymis weights or testis or thyroid histopathology.

Complete clinical chemistry data are also available for multiple timepoints in the chronic rat and

dog studies, as is additional thyroid weight and histology from chronic studies in rats and dogs.

The results from all of these studies, in particular the two-generation rat reproduction studies,

developmental neurotoxicity study (Sheets et al., 2008) and 2007 publication by Goetz et al.

should be considered adequate to address the data requirement for a male pubertal assay.

Therefore, the need to conduct this assay should be waived.

In vivo Hershberger Assay

The Hershberger assay is a short-term in vivo assay to detect androgenic or antiandrogenic

chemicals or chemicals that inhibit 5a-reductase by measuring weight changes in five

androgen dependent tissues: ventral prostate (VP), seminal vesicle (SV) (plus fluids and

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coagulating glands), levator ani-bulbocavernosus (LABC) muscle, paired Cowper's glands

(COW) and the glans penis (GP). Sufficient data on these specific tissues are not available

in guideline or in research studies reported in the public domain, and therefore the Consortium

will conduct the Hershberger assay.

Conclusion for AR Assays

Adverse effects that would be expected from exposure to an androgen disrupting chemical are

shown below. The effects are not consistent with what has been observed in mammalian

toxicology studies with triadimefon.

Mechanism Potential Effects in Males Potential Effects in Females

Androgenicity Increased or decreased male

reproductive organ weights

Reduced sperm counts

Testicular atrophy

Increased anogenital distance

Accelerated vaginal opening

Reduced fertility

Altered differential follicle count

Histopathological findings of the

female reproductive organs

(e.g., vaginal agenesis)

Induced male sex accessory

tissues

Anti-androgenicity Delayed preputial separation

Reduced anogenital distance

Ectopic testes

Hypospadias/epispadias

Reduced fertility

Reduced reproductive organ weight

(i.e., prostate, seminal vesicles)

Retained nipples/areolas

Histopathological findings of the

reproductive organs (e.g., epididymal

agenesis, testicular tumors)

• Altered pup sex ratios between

external and internal sexing

In summary, scientific information indicates that an androgenic compound would produce

among other findings changes in sexual development, estrous cyclicity, fertility and other

reproductive endpoints, as well as to produce weight and histopathological changes to

reproductive and sexual accessory organs. Adequate information on triadimefon effects on most

parameters is available in existing regulatory studies and publications. Given the level of

available information, it is considered unnecessary to conduct either the androgen receptor

binding assay or to sacrifice the additional animals needed to conduct the male pubertal in vivo

androgen screening assays. In order to address uncertainties regarding organ weight effects

and increased testosterone levels reported in the Goetz publication, a Hershberger assay will be

conducted.

Thyroid System OSRI

Assays (rat pubertals and amphibian metamorphosis) have been included in the EDSP Tier 1

screening battery to assess the potential of a chemical to interfere with thyroid function.

Although these assays as specifically described in EPA's OPPTS 890 Guideline Series have not

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been conducted with triadimefon, adequate information exists to assess triadimefon potential to

interfere with the hypothalamic-pituitary-thyroidal (HPT) axis. As described previously,

information on thyroid weight and histology are available from guideline studies including

lifetime feeding studies in rats and mice, and a chronic dog study as well as a number of

publications by EPA researchers. Rockett et al. (2006) and Goetz et al. (2007) evaluated

triadimefon effects on thyroid weight and histopathology in developing female and male rats,

respectively, exposed in utero, throughout lactation, and up to PND 92 for males and PND 98

for females. In addition, thyroid hormone level measurements at various postnatal ages have

been conducted by EPA researchers (Goetz et al., 2007, Tully et. al, 2006 both previously

discussed and Wolf et al., 2006 discussed below).

No histopathological changes were observed in dog or mouse thyroid (or pituitary) glands in the

long term regulatory studies discussed earlier (Bombard, 1986, Hoffman and Groning, 1978). In

the rat lifetime feeding study (Bomhard and Schilde, 1991), histopathologic changes were

observed in the thyroid in rats administered triadimefon in the diet at 1800 ppm:114/199

(male/female) mg/kg bw/day (HOT), but only in animals sacrificed at study termination, i.e. after

approximately 2 years of exposure; no effects on the thyroid were observed in animals that were

sacrificed after 1 year. The changes consisted of an increased incidence of cystic hyperplasia

and an increased incidence of thyroid adenomas in both sexes, which were suggested by BCS

to be within historical control range (Sander, E. and Schilde, B., 1993). EPA's Cancer Peer

Review Committee concluded that the carcinogenic potential of triadimefon is equivocal based

on these findings.

In the 2006 study by EPA researchers Wolf et al., the triadimefon mode of action for thyroid

tumorigenesis was investigated. Seven week old Wistar Han male rats received triadimefon in

the diet at concentrations of 100, 500 or 1800 ppm for 4, 30 or 90 days (13 animals per dose

group). Thyroid glands (among others) were taken at necropsy and scored for colloid depletion,

follicular cell hypertrophy and/or follicular cell hyperplasia. Total circulating serum thyroid

hormone levels were analyzed as well as cholesterol, triglycerides and high density lipoproteins.

Hepatic enzyme activity was assessed. The histology of the thyroid was only different from

control in rats treated with the high dose of triadimefon for 30 days. These rats had colloid

depletion and follicular cell hypertrophy, which were not different from control for rats treated for

90 days. From hormone level and enzyme activity findings the authors concluded that although

triadimefon induces UDPGT activity and reduces serum T4 levels, there is no associated

increase in TSH levels or persistently increased thyroid follicular cell proliferation. However,

BCS suggests that the fact that a compensatory TSH increase was not seen may have been

because the impact on the thyroid over the duration of dosing was not severe enough or the

timing of blood sampling was not optimal. Based upon the finding of increased UDPGT activity,

the alteration of thyroid hormone balance secondary to liver induction remains a plausible

mechanism.

Other downstream effects which might be indicative of interference of the thyroid system include

changes in endpoints such as acoustic startle and motor activity in the DNT study and effects on

fetal/pup weight and survival. No effects on any of these parameters were observed in the

offspring of dams administered diet containing triadimefon at levels of 100 and 300 ppm from

gestation day 6 through lactation day 21 (Sheets et al., 2008). At 800 ppm, pre-weaning body

weight and body weight gain were reduced for high dose pups for both sexes and there was an

increased peak amplitude during auditory startle reflex testing in high dose females considered

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equivocal. In developmental toxicity studies in rats (Unger, 1982, Nagumo et al., 1981) and

rabbits (Clemens and Hartnagel, 1990), there were no treatment-related effect on gestation or

fertility indices at the highest doses tested. Rat fetal weights were unaffected by treatment;

rabbit fetal weights were slightly reduced in high dose pups.

Adverse effects that would be expected from exposure to a thyroid function disrupting chemical

are shown below.

Mechanism Potential Effects in Males Potential Effects in Females

Thyroid hormone

modulation

Decreased T3 and/or T4 levels, increased TSH levels

Increased thyroid weights

Decreased colloid content and thyroid follicle size

Follicular cell hypertrophy

Thyroid follicular cell hyperplasia

Thyroid follicular cell tumors

The Amphibian Metamorphosis Assay (AMA) involves the use of tadpoles to determine if

chemicals affect the hypothalamic-pituitary-thyroid (HPT) axis during metamorphosis and

consequently results in developmental effects (USEPA, 2009a). The AMA represents a

generalized vertebrate model to the extent that it is based on the conserved structure and

function of the HPT axis (USEPA 2009b). A review of existing thyroid assays was reported by

OECD in a detailed review paper (OECD 2006). This report points out the conserved nature of

the HPT axis. As outlined above, a number of studies have been conducted to investigate the

potential effects of triadimefon on thyroid systems of vertebrates. These studies were also done

with conserved models so an additional evaluation with an additional species, with the highly

conserved HPT axis, is unnecessary to evaluate the thyroid activity potential in vertebrates.

The substantial existing data from studies should be deemed adequate for a screening level

thyroid evaluation. Therefore, the conduct of an AMA is unnecessary and would result in the

expenditure of resources and unnecessary sacrifice of animals.

Results of guideline and published studies indicate that repeated exposure to triadimefon at

sufficient doses does result in microscopic changes in the rat thyroid. Adequate information on

triadimefon effects on all parameters is available in existing regulatory studies and publications.

Given the level of available information, it is considered unnecessary to conduct either of the rat

pubertal or the amphibian metamorphosis assays. Therefore, the requirement for the thyroid

function Tier 1 assays should be waived.

Steroid Synthesis OSRI

Triadimefon was included in EPA's aromatase assay validation study and results are reported in

EPA's December 11, 2007 "Integrated Summary Report on Aromatase"" and fulfill the

requirement for the aromatase assay.

The endpoints discussed above for the estrogen and androgen systems are also relevant for

assessment of a compound's potential to interfere with steroidogensis. These endpoints are

shown in Table 1a and those considered as functionally equivalent to address the purpose of

the steroidogenesis assays are indicated. In addition, a study by EPA researchers (Goetz, et

al., 2009) reports results of in vitro assays in rat testis and H295R cells as well as in vivo serum

and testis testosterone levels in male rats after 14 or 30 days dietary exposure.

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Regarding the in vitro assay, only 5 rather than 7 concentrations were tested in the H295R cell

assay, and only 1 control was used instead of the 2 specified for OPPTS 890.1550. In addition

to testosterone and estradiol, progesterone levels were also measured. The author concluded

that triadimefon is a weak inhibitor of testosterone production in vitro. This stands in contrast to

the in vivo findings were testosterone levels were elevated (discussed below). The published

study is considered adequate to fulfill the requirement for the in vitro steroidogenesis assay.

Regarding the in vivo investigation, male Wistar han rats (PND 60) were administered

triadimefon in the diet at 1800 ppm for 30 days. Body weight and food consumption were

measured weekly; animals were tail bled on day 14 for testosterone measurements. On day 30

animals were decapitated and trunk blood collected for serum measurements for testosterone,

estradiol, LH and prolactin. Testes were weighed and snap frozen; liver, epididymis, ventral

prostate, seminal vesicles and pituitary were collected and weighed. Body weights were lower in

treated rats relative to control throughout the course of the study. Serum testosterone levels

were not increased after 14 days but were significantly increased at 30 days (ca. 290% change),

as was intra-testicular testosterone (210% change). Serum levels of estradiol, LH and prolactin

were somewhat elevated at 30 days, but increases were not statistically significant. Liver

weights, both absolute and relative, were significantly increased after 30 days of exposure (27

and 37%, respectively). Absolute pituitary and paired epididymal weights, but not relative, were

reported to be somewhat decreased (9.8 and 5.2%, respectively, p<0.05). Androgen dependent

ventral prostate and seminal vesicle weights were unaffected, which the author states are

consistent with other studies which do not show much evidence of AR antagonism in androgendependent tissues (citing Goetz, et al., 2007; Tully et al., 2006 ; Okubo et al., 2004).

Goetz et al. conclude that triadimefon appears to inhibit CYP17A1 in vitro, decreases

testosterone levels in vitro, but increases rat serum and testis testosterone levels in vivo after

sufficient duration of exposure by an unresolved mechanism.

Adverse effects that would be expected from exposure to a chemical inhibiting steroidgenesis or

aromatase are shown below.

Mechanism Potential Effects in Males Potential Effects in Females

Reduced steroid

biosynthesis

Similar to anti-androgenicity

Possible increased serum

cholesterol levels

Increased Leydig cell tumors

Similar to anti-estrogenicity/

aromatase inhibition

Aromatase

inhibition

Increased time to mating

Decreased male mounting behavior

Decreased body weight (chronic)

Increased testis weight (chronic)

Increased body weight

Decreased uterine weight

Increased ovary size

Polycystic ovaries

Stromal hyperplasia in ovary

(chronic)

Hyalinization in ovary (chronic)

Increased ureter and bladder

infection

Decreased mammary tumor

incidence (in Sprague Dawley rats)

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Given the level of available information from US EPA published research, the requirement for

aromatase and steroidgenesis assays is fulfilled.

Overall Conclusion

The Triadimefon Consortium believes that existing data on triadimefon are for the most part

adequate to assess the potential for this compound to interact with estrogen, androgen and

thyroid systems. However, in order to address equivocal findings and supply additional studies

that will provide complementary data and sufficient scientific redundancy to assure that

sufficient definitive data are available to perform a weight of evidence (WOE) evaluation of

whether any Tier 2 testing would be necessary, the Triadimefon Consortium will conduct a Fish

Short term Reproduction Assay and a Hershberger Bioassay. The Triadimefon Consortium

requests waivers from all other Tier 1 studies as justified below.

Endocrine Disruptor Screening Program Tier 1

Test

Justification

In Vitro Screening Assays

OPPTS 890.1250: Estrogen Receptor Binding Assay

Using Rat Uterine Cytosol (ER-RUC)

OPPTS 890. 1300: Estrogen Receptor

Transcriptional Activation (Human Cell Line (HeLa9903))

OPPTS 890.1150: Androgen Receptor Binding

(Rat Prostate Cytosol)

OPPTS 890. 1200: Aromatase

(Human Recombinant)

OPPTS 890. 1550: Steroidogenesis

(Human Cell Line -H295R)

Functional equivalence of OSRI

Functional equivalence of OSRI

Functional equivalence of OSRI

Included as a test substance in EPA's validation

of the assay

Functional equivalence of OSRI plus publication

assessed as sufficiently equivalent to protocol

In Vivo Screening Assays

OPPTS 890. 1600: Uterotrophic Assay

OPPTS 890.1450: Pubertal Development and

Thyroid Function in Intact Juvenile/ Peripubertal

Female Rats

OPPTS 890.1500: Pubertal Development and

Thyroid Function in Intact Juvenile/ Peripubertal Male

Rats

OPPTS 890.1100: Amphibian Metamorphosis (Frog)

Functional equivalence of OSRI

Functional equivalence of OSRI

Functional equivalence of OSRI

Functional equivalence of OSRI

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REFERENCES

The following studies have been submitted previously to EPA and are cited by MRID number:

MRID No. 32539 Hoffman, K. and Groning, P. (1978) Meb 6447 Long-Term Toxicity Study on

Dogs (Two-Year Feeding Study): Report No. 7882; Report No. 66656. (Unpublished study

received Apr 28, 1980 under 3125-318; prepared by Bayer, AG, submitted by Mobay Chemical

Corp., Kansas City, Mo.; CDL099413-D).

MRID No. 32541 Loser, E. and Lorke, D. (1979) Meb 6447 Multigeneration Reproduction

Study on Rats: Report No. 8297; Report No. 67752. (Unpublished study received Apr 28, 1980

under 3125-318; prepared by Bayer, AG, submitted by Mobay Chemical Corp., Kansas City,

Mo.;CDL: 099413-F)

MRID No. 48624 Mohr, U. (1976) Meb 6447: Subchronic Toxicity Study on Rats (Twelve -

Week Feeding Experiment): Report No. R 840 a; Report No. 44300. (Unpublished study

received Aug 15, 1977 under 3125-318; prepared by Medizinische Hochschule Hannover,

Abteilung fur Experi- mentelle Pathologie, West Germany, submitted by Mobay Chemical Corp.,

Kansas City, Mo.; CDL231312-C)

MRID No. 48625 Hoffman, K.and Luckhaus, G. (1974) Meb 6447: Subchronic Toxicity Study

on Dogs (Thirteen-Week Feeding Experiment): Report No. 5071; Report No. 44737.

(Unpublished study received Aug 15, 1977 under 3125-318; prepared by Bayer, AG, submitted

by Mobay Chemical Corp., Kansas City, Mo.; CDL231312-D).

MRID No. 60226 Hoffmann, K., Luckhaus, G. and Lorke, D. (1974) Meb 6447: Subchronic

Toxicity Study on Dogs (Thirteen-Week Feeding Experiment): Report No.5071; Report No.

44737. (Unpublished study received Dec 15, 1977 under 3125-EX-149; prepared by Bayer, AG,

W. Germany, submitted by Mobay Chemical Corp., Kansas City, Mo.; CDL: 232490-D).

MRID No. 89023 Nagumo, K., Teraki, Y. and Chiba, T. (1981) Teratogenicity

Test of MEB 6447 in Pregnant Rats: Submitter 80257. (Unpublished study received Jan 5,

1982 under 1F2474; prepared by St. Marianna Univ., First Dept. of Anatomy, Laboratory of

Embryology, submitted by Mobay Chemical Corp., Kansas City, Mo.; CDL: 070570-A)

MRID No. 126261 Hoffmann, K., Groning, P. and Lorke, D. (1978) MEB 6447: Long-term

Toxicity Study on Dogs (Two-year Feeding Study): Report No. 7882; 66656. (Unpublished study

received Mar 11, 1983 under 3125-346; prepared by Bayer AG, W. Ger., submitted by Mobay

Chemical Corp., Kansas City, MO; CDL:071468-B)

MRID No. 149336 Unger, T., Van Goethem, D. and Shellenberger, T. (1982) A Teratological Evaluation of Bayleton in Mated Female Rats: MRI Project No. 7272-B. Unpublished

study prepared by Midwest Research Institute. 52 p.

MRID No. 155075 Eiben, R. (1984) MEB 6447 (Triadimefon): Two-generation Study with

Rats: Supplementary Study: Report No. 12712. Unpublished study prepared by Bayer AG,

Institute of Toxicology. 168 p.

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MRID No. 40752101 Bombard, E. (1986) MEB 6447: Carcinogenicity Study on NMRI Mice (21-

Month Administration in the Feed): Report No. 87287. Unpublished study prepared by Bayer

AG. 1190 p.

MRID No. 40865101 Bombard, E. and Hahnemann, S. (1986) MEB 6447: Common Name:

Triadimefon, the Active Ingredient of Bayleton: Carcinogenicity Study on NMRI

Mice: Project ID 87287. Unpublished study prepared by Bayer Ag. 5 p.

MRID No. 41446201 Clemens, G. and Hartnagel, R. (1990) Teratology Study in the Rabbit

with MEB 6447 (Triadimefon): Report No.: MTD0149. Unpublished study prepared by Miles,

Inc. 155 p.

MRID No. 42089601 Clemens, G.; Hartnagel, R. (1991) Teratology Study in the Rabbit

with MEB 6447: Supplement to MRID 41446201: Lab Project Number: 100026: MTD0149.

Unpublished study prepared by Miles, Inc., Tox. Dept. 11 p.

MRID No. 42153901 Bomhard, E.; Schilde, B. (1991) MEB 6447: Chronic Toxicity and

Cancerogenicity Studies on Wistar Rats with Administration in Diet over a Period of 105 Weeks:

Lab Project Number: 20774: 101922. Unpublished study prepared by Bayer Ag., Dept. of

Toxicology. 1029 p. Also see MRID No. 43089301

MRID No. 43089301 Sander, E. and Schilde, B. (1993) Supplement: The Incidence of

Thyroid Follicular Cell Tumors in Wistar Rats: (BAYLETON MEB 6447): A Compilation of

Historical Data: Lab Project Number: T/3027626: 20774/A. Unpublished study prepared by

Bayer AG. 57 p.

MRID No. 47377101 Sheets, L.; Gilmore, R.; Hoss, H. (2008) A Developmental Neurotoxicity

Study with Technical Grade Triadimefon in Wistar Rats. Project Number: 07/D72/IL, 201766.

Unpublished study prepared by Bayer CropScience. 1179 p.

MRID No. 92188018 Eigenberg, D. (1990) Mobay Corporation Phase 3 Summary of MRID

00149336. A Teratological Evaluation of Bayleton (Triadimefon)in Mated Female Rats: Project

7272-B (Mobay Report 82270). Prepared by MIDWEST RESEARCH INSTITUTE. 22 p.

MRID No. 92188019 Eigenberg, D. (1990) Mobay Corporation Phase 3 Summary of MRID

00155075 and Related MRIDs 00032541. MEB 6447 (Triadimefon) Multigeneration

Reproduction Study on Rats; (Triadimefon) Two-generation Study with Rats (Bayleton): Projects

8297 and 12712 (Mobay Reports 67752 and 82270). Prepared by BAYER AG. 38 p.

MRID No. 92188020 Loser, E. (1990) Mobay Corporation Phase 3 Reformat of MRID

00032541 and Related MRIDs 00155075. MEB 6447 (Triadimefon) Multigeneration

Reproduction Study on Rats: Project Number 67752; Report No. 8297. Prepared by BAYER

AG. 173 p.

The following guidelines and review paper were used for reference:

OECD, 2006. Draft Detailed Review Paper on Thyroid Disruption Assays. OECD Environment,

Health and Safety Publication Series on Testing and Assessment. No. not assigned. Paris,

France.

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USEPA, 2009a. Test Order/DCI #: EDSP-109901-156 for Triadimefon. Date Issued:

12/17/2009.

USEPA, 2009b. Office of Prevention, Pesticides and Toxic Substances. Test Guideline,

890.1100: Amphibian Metamorphosis (Frog). EPA 740-C-09-002. October 2009. U.S.

Environmental Protection Agency. Washington, D.C.

Other Published literature:

Goetz, A., Ren, H., Schmid, J., Blystone, C., Thillainadarajah, I., Best, D., Nichols, H., Strader,

L, Wolf, D., Narotsky, M., Rockett, J. and Dix, D. (2006). Toxicol. Sci. 95(1): 227-239.

Goetz, A., Rockett, J., Ren, H., Thillainadarajah, I. and Dix, D. (2009). Syst. Biol. Reproof. Med.

55: 214-226.

Klimisch, H.-J., Andreae, M and Tillman, U. (1997). A Systematic Approach for Evaluating the

Quality of Experimental Toxicological and Ecotoxicological Data. Regulatory Tox. and Pharm.

25: 1-5.

Kojima, H., Katsura, E., Takeuchi, S., Niiyama, K., and Kobayashi, K. (2004). Screening for

estrogen and androgen receptor activities in 200 pesticides by in vitro reporter gene assays

using Chinese Hamster Ovary cells. Environ. Health Perspect. 112: 524-531.

Okubo, T., Yokoyamo, Y., Kano, K., Soya, Y. and Kano, I. (2004). Arch. Environ. Contam.

Toxicol. 46: 445-453.

Petit, F., Le Goff, P., Crevadi, J-P., Valotaire, T., and Pakdel, F. (1997). Two complementary

bioassays for screening the estrogenic potency of xenobiotics: recombinant yeast for trout

estrogen receptors and trout hepatocyte cultures. J. Mol. Endocrin. 19: 321-335.

Rockett, J., Narotsky, M., Thompson, K., Thillainadarajah, I., Blystone, C., Goetz, A., Ren, H.,

Best, D., Murrell, R., Nichols, H., Schmid, J., Wolf, D. and Dix, D. (2006). Effect of conazole

fungicides on reproductive development in the female rat. Reprod. Toxicol. 22: 647-658.

Tully, B., Bao, W., Goetz, A., Blystone, C., Ren, H., Schmid, J., Strader, L., Wood, C., Best, D.,

Narotsky, M., Wolf, D., Rockett, J. and Dix, D. (2006). Gene expression profiling in liver and

testis of rats to characterize the toxicity of triazole fungicides. Toxicol. Appl. Pharmacol. 215:

260-273.

Vinggaard, A.M., Breinholt, V. and Larsen, J.C. (1999). Screening of selected pesticides for

oestrogen receptor activation in vitro. FoodAddit. Contam. 16(12): 533-542.

Wolf, D., Allen, J., George, M., Hester, S., Sun, G., Moore, T., Thai, S-H., Delker, D., Winkfield,

E., Leavitt, S., Nelson, G., Roop, B., Jones, C., Thibodeaux, J. and Nesnow, S. (2006). Toxicity

profiles in rats treated with tumorigenic and non-tumorigenic triazole conazole fungicides:

propiconazole, triadimefon and myclobutanil. Toxicologic. Pathology 34: 895-902.

Page 22 of 46
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Table 1a: Functional Equivalence Matrix - Apical Endpoints in Existing Tridimefon Studies

Relevant to Evaluation of Potential Effects Resulting from Modes of Action Assessed in the In

Vitro Assays

OPPTS 870.3800 Two-Generation Rat

Reproduction

(combined MRIDs 0032541, 155075)

• Reproductive indices (FO and F1

parents)

• Litter parameters (F1 and F2J

• Pup Sex Ratio

• External genitalia anomalies (F1

and F2}

• Organ wts ovaries (FO and F1

parental 9)

• Organ histopathology: uterus,

ovaries, vagina (F2 pups only) (F1

parental, F2 and F3 pups 9)

• Organ wts: epididymides + testes

(FO and F1 parental CT)

• Organ histopathology: seminal

vesicles and prostate (F1 parental

only), epididymides, testes (F1

parental, F2 and F3 pups C?)

• Organ histopathology: pituitary (F1

parental and F2 pups) adrenals (F1

parental, F2 and F3 pups) and

thyroid (F3 pups)

• Ovary follicular status at necropsy

(F1 parental, F2 and F3 pups §)

• Andrological parameters (sperm

count, motility, production and

anomalies - FO and F1 parental C?)

OPPTS 870.6300 Developmental

neurotoxicity (MRID 473771 01)

• Pup Sex Ratio

• Vaginal Opening

• Preputial Separation

• External genitalia anomalies

OPPTS 870.3100 Subchronic toxicity in

rats (MRID 48624)

• Organ wts and histopathology:

ovaries

• Organ wts and histopathology:

ER binding

and

transcriptional

activation

^

S

^

•/

</

V

.

-

V

-

V

s

-

s

s

AR

binding

S

S

s

s

-

-

s

</

-

s

s

I

^

h

 ^

.

Aromatase

S

s

s

s

s

s

</

s

V

s

V

s

s

s

s

V

Steroidogenesis

V

V

s

s

V

s

s

s

s

s

s

s

s

s

s

V

Page 23 of 46
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testes

• Organ wts and histopathology:

adrenal, pituitary, thyroid

OPPTS 870.3150 Subchronic toxicity in

dogs (MRID 48625, 60226)

• Organ wts and histopathology:

ovaries, uterus (histopath only)

• Organ wts and histopathology:

prostate, testes, epididymides

(histopath only)

• Organ wts and histopathology:

adrenal, pituitary, thyroid

OPPTS 870.41 OOb Chronic toxicity in

dogs (MRIDs 32539, 126261)

• Organ wts and histopathology:

ovaries; histopath only mammary

glands, uterus

• Organ wts and histopathology:

prostate, testes; histopath only

seminal vesicles, epididymides

• Organ wts and histopathology:

adrenal, pituitary, thyroid

OPPTS 870.4200 Mouse carcinogenicity (MRID 40752101, 40865101)

• Organ histopathology: mammary

glands, uterus, ovaries, vagina

• Organ histopathology: seminal

vesicles, epididymides, prostates,

testes

• Organ histopathology: adrenal,

pituitary, thyroid

OPPTS 870.4300 Rat combined toxicity/carcinogenicity (MRID 42153901 )

• Organ histopathology: mammary

glands, uterus, ovaries, vagina

• Organ wts and histopathology:

testes; histopath only seminal

vesicles, epididymides, prostate

• Organ wts and histopathology:

adrenal,; histopath only pituitary,

thyroid

ER binding

and

transcriptional

activation

-

-

S

-

-

V

-

-

s

-

-

V

-

AR

binding

S

-

.

s

-

-

V

-

m

s

-

_

V

Aromatase

V

S

S

V

V

s

s

s

V

s

s

V

s

s

Steroidogenesis

^

v'

S

S

S

V

S

S

s

s

s

s

s

s

Page 24 of 46
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Table 1b: Functional Equivalence Matrix - Apical Endpoints in Existing Triadimefon Studies

Relevant to Parameters Assessed in the In Vivo Assays

OPPTS 870.3800 Two-Generation Rat

Reproduction

(combined MRIDs 0032541, 155075)

• Growth (body weight)

• Reproductive indices (FO and F1

parents)

• Pup Sex Ratio

• External genitalia anomalies (F1 and

F2)

• Organ wts: ovaries (FO and F1

parental 9)

• Organ histopathology: uterus,

ovaries, vagina (F2 only) (F1 parental

and F2and FSpups 9)

• Organ wts: seminal vesicles

epididymides + testes ( FO and F1

parental cf)

• Organ histopathology: seminal

vesicles (with coagulating glands and

their fluids), epididymides (with

contents), prostate, testes (F1

parental and F2 and F3 pups cf)

• Organ wts: adrenal and pituitary (FO

and F1 parents)

• Organ histopathology: adrenal and

pituitary (F1 parents and F2pups)

• Andrological parameters (sperm

count, motility, production and

anomalies - FO and F1 parental C?)

OPPTS 870.6300 Developmental

neurotoxicity (MRID 473771 01J

• Pup Sex Ratio

• Vaginal Opening

• Preputial Separation

• External genitalia anomalies

OPPTS 870.3100 Subchronic toxicity in

rats (MRID 0048624)

• Organ wts and histopathology:

ovaries

• Organ wts and histopathology: testes

• Organ wts and histopathology:

adrenal, pituitary, thyroid

Pubertal

Female

</

S

S

s

s

V

-

-

s

s

-

s

s

-

s

V

.

-

Pubertal

Male

^

v'

V

S

-

-

S

S

v'

S

S

V

-

s

</

.

V

-

Uterotrophic

-

/

•/

V

S

V

V

S

s

s

-

s

s

-

s

V

s

s

Hershberger

-

</

S

V

S

V

s

s

s

V

s

V

-

s

s

s

s

s

Page 25 of 46
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OPPTS 870.3150 Subchronic toxicity in

dogs (MRID 0048625, 60226)

• Organ wts and histopathology:

ovaries, uterus (histopath only)

• Organ wts and histopathology:

prostate, testes, epididymides

(histopath only)

• Organ wts and histopathology:

adrenal, pituitary, thyroid

OPPTS 870.41 OOb Chronic toxicity in dogs

(MRID 32539, 126261)

• Organ wts and histopathology:

ovaries; histopath only mammary

glands, uterus

• Organ wts and histopathology:

prostate, testes; histopath only

seminal vesicles, epididymides

• Organ wts and histopathology:

adrenal, pituitary, thyroid

OPPTS 870.4200 Mouse carcinogenicity

(MRID 40752101, 40865101)

• Organ histopathology: mammary

glands, uterus, ovaries, vagina

• Organ histopathology: seminal

vesicles, epididymides, prostates,

testes

• Organ histopathology: adrenal,

pituitary, thyroid

OPPTS 870.4300 Rat combined toxicity/carcinogenicity {MRID 4291 53901)

• Organ histopathology: mammary

glands, uterus, ovaries, vagina

• Organ wts and histopathology: testes;

histopath only seminal vesicles,

epididymides, prostate

• Organ wts and histopathology: adrenal; histopath only pituitary, thyroid

•

Pubertal

Female

S

-

-

s

-

s

s

-

V

s

-

s

Pubertal

Male

_

V

-

-

S

S

_

^

s

_

V

V

Uterotrophic

V

S

S

s

s

-

s

-

-

s

-

•/

Hershberger

^

S

S

s

s

-

.

s

-

m

V

s

Page 26 of 46
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Table 2: Female pubertal assay endpoints compared to reported endpoints for triadimefon from

multigeneration rat reproduction studies (MRIDs 32541,155075), a developmental neurotoxicity

study (MRID 4737101) and relevant findings from publications "Effect of conazole fungicides on

reproductive toxicity in the female rat" (Rockett et.al., Reproductive Toxicology 22 (2006) 647-658)

Female Pubertal

Assay

Multigeneration Rat Reproduction Studies

MRID 32541 MRID 155075

Rockett et. al.

Reproductive Toxicology

22 (2006)

Exposure Duration

PND 22 through 42

for females

Dose levels (diet)

FO animals - throughout 10 weeks (32541) or 14

weeks (155075) premating, mating, gestation,

and lactation periods

F1 animals - in utero (as fetus) and lactation

periods (as neonates) followed by exposure

during 10 (32541) or 14 (155075) weeks

premating, mating, gestation and lactation

periods for F1 parents

F2 animals - in utero (as fetus) and lactation

periods (as neonates)

F3 animals as for F2 (32541 )

0,50,300,1800 0,50,1800

Mated dams -from GD6 -

LD21

F1 females - in utero (as

fetus) and lactation periods

as neonates) and from

PND 22 through 98

0, 100, 500, 1800

Endpoints Included

Daily body weight

Age and body weight

at sexual

development (i.e.

vaginal opening*)

Organ weights

• Uterus (blotted)

• Ovaries (paired)

• Thyroid (after

fixation)

• Liver

• Kidneys (paired)

• Pituitary

• Adrenals (paired)

Histology

• Uterus

• Ovaries

• Thyroid (colloid

area and follicular cell

height)

• Kidney

Estrous cyclicity

• Age at first vaginal

estrus after vaginal

Weekly body weight

(all animals)

Histology F3 pups at

weaning (PND28)

• Brain

• Uterus

• Ovaries

• Liver

• Kidneys

• Adrenals

• Thyroid

Weekly body weight (all

animals)

Organ weights (FO, F1

parents)

• Brain

• Ovaries

• Liver

• Kidneys

Histology F1 parent, F2

pups at weaning (PND28)

• Brain

• Uterus

• Ovaries

• Pituitary

• Liver

• Kidneys

• Adrenals

• Vagina (F2 only)

Not clearly reported

Anogenital distance at

PNDO; bodyweight not

shown

Age at VO; body weight

not shown

Organ weights (relative

reported PN 99)

Brain

Hypothalamus

Uterus (drained)

Ovaries (paired)

Liver

Others taken and

weighed but not reported in

publication: hippocampus,

pituitary, thyroid

Histology

• Liver

• Thyroid (colloid depletion

and follicular cell

hypertrophy and/or

hyperplasia)

• Ovaries

Estrous cyclicity

Percent of females experiencing regular, extended

Page 27 of 46
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opening

• Length of cycle

• Percent of animals

cycling

• Percent of animals

cycling regularly

Clinical (serum)

chemistry

• Standard blood

panel, including

creatinine and blood

urea nitrogen

Thyroid hormones

Data not available from reproduction study but

clinical chemistry data are available from multiple

timepoints in chronic rat and dog studies

or abnormal cycles during

weeks 1-2, 5-6 or 9-10

after VO

Serum estradiol at at PND

99

Measured in male cohort

study, see Table 3

*reported in the triadimefon guideline developmental neurotoxicity study

Page 28 of 46
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Table 3: Male pubertal assay endpoints compared to reported endpoints for triadimefon from

multigeneration rat reproduction studies (MRIDs 32541,155075), a developmental neurotoxicity

study (MRID 4737101) and relevant findings from publication "Disruption of testosterone

homeostasis as a mode of action for the reproductive toxicity of triazole fungicides in the male

rat" (Goetz et.al., Toxicological Sciences 95(1), 227-239 (2007).

Male Pubertal

Assay

Exposure Duration

PND 23 through 53

for males

Dose levels (diet)

Multigeneration Rat Reproduction Studies

MRID 32541 MRID 155075

FO animals -throughout 10 weeks (32541) or 14

weeks (155075) premating, mating, gestation, and

lactation periods

F1 animals - in utero (as fetus) and lactation

periods (as neonates) followed by exposure during

10 (32541) or 14 (155075) weeks premating,

mating, gestation and lactation periods for F1

parents

F2 animals - in utero (as fetus) and lactation

periods (as neonates)

F3 animals as for F2 (32541 )

0, 50, 300, 1800 0,50,1800

Goetz et. al.

Reproductive

Toxicological Sciences

95(1) (2006)

Mated dams -from GD6

-LD21

F1 males - in utero (as

fetus) and lactation

periods (as neonates)

and from PND 22

through 120

0,100,500,1800

Endpoints Included

Daily body weight

Age and body weight

at sexual development (i.e. preputial

separation*)

Organ weights

• Seminal vesicle

plus coagulating

glands (with and

without fluid)

• Ventral prostate

• Dorsolateral

prostate

• Levator

ani/bulbocavernosus

muscle complex

• Epididymides (left

and right separately)

• Testes (left and

right separately)

• Thyroid (after

fixation)

• Liver

• Kidneys (paired)

• Pituitary

• Adrenals (paired)

Histology

• Epididymis (one)

Weekly body weight (all

animals)

Histology F3 pups

• Brain

Weekly body weight (all

animals)

Organ weights (FO, F1

parents)

• Brain

• Testicles

• Liver

• Kidneys

Histology F1 parents, F2

pups at weaning

Weekly from birth

Anogenital distance and

body weight at PNDO;

Age and body weight at

PPS

Organ weights, rel and

abs(PND1,22, 50, 92, 1

male/litter)

• Liver" ***

• Testes**

• Ventral prostate**

• Epididymis**

•Seminal vesicle**

•Brain**

•Pituitary

• Others taken and

weighed but not

reported in publication:

hypothalamus,

hippocampus, thyroid**

Histology

• Liver** ***

Page 29 of 46
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• Testis (one)

• Thyroid (colloid

area and follicular

cell height)

• Kidney

Clinical (serum)

chemistry

• Standard blood

panel, including

creatinine and blood

urea nitrogen

Thyroid hormones

• Testes

• Epididymis

• Liver

• Kidneys

• Adrenals

• Thyroid

(PND28)

• Brain

• Testes

• Epididymis (F1 only)

• Prostate (F1 only)

• Pituitary

• Liver

• Kidneys

• Adrenals

Data not available from reproduction study but

clinical chemistry data are available from multiple

timepoints in chronic rat and dog studies

Serum testosterone,

Follicular stimulating

hormone

Sperm morphology &

motility

• Testes**

• Ventral prostate

• Thyroid** ***

• Epididymis

• Pituitary

T3, T4, TSH***

Serum testosterone**,

Estradiol, Luteinizing

hormone** at PND50

and 92

Sperm morphology &

motility**

*reported in the triadimefon guideline developmental neurotoxicity study

** also examined in Tully, et.al., in 2006

*** also examined in Wolf, et.al., 2006

Page 30 of 46
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Table 4: Functional Equivalence Matrix - Apical Endpoints in Existing Studies and Publications

Relevant to Amphibian Metamorphosis

Guideline Studies and Publications

Multi-Generation Rat Reproduction

(MRID 0032541 and 155075)

• F1 and F2 pup survival and growth

• 4 wk old F3b pups histopathology

thyroid, adrenals (MRID 0032541)

Developmental toxicity (MRIDs)

• Fetal survival and weight

Developmental neurotoxicity

(MRID)

• Pup survival and growth

• Acoustic Startle

• Motor Activity

• Surface righting

Goetz et. al. Reproductive

Toxicological Sciences 95(1) (2006)

• Pup survival and growth (bw)

• Serum hormone levels T3, T4,

TSH

• Organ wts and histopathology

thyroid, pituitary

Rat subchronic toxicity (MRID

• Organ wts and histopathology:

adrenal, pituitary, thyroid

Wolf et.al., Toxicologic Pathology, 34

(2006)

• Male rat growth from 7 wks (bw)

• Serum hormone levels T3, T4,

TSH (ages 53, 79, 139d)

• Histopathology thyroid (ages as

above)

Dog subchronic toxicity (MRID

• Organ wts and histopathology:

adrenal, pituitary, thyroid

Rat combined toxicity/carcinogenicity (MRID)

• Organ wts and histopathology:

adrenal, pituitary, thyroid

Chronic toxicity in dogs (MRID)

• Organ wts and histopathology:

adrenal, pituitary, thyroid

Mouse carcinogenicity (MRID)

• Organ wts and histopathology:

adrenal, pituitary, thyroid

Amphibian Metamorphosis

Developmental Stage,

Length and Weight

Measurements

</

•/

•/

S

s

s

s

•/

Thyroid Gland

Histology

</

~j

s

s

s

s

s

s

s

</

Page 31 of 46
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APPENDIX A

TITLE PAGES WITH MRID NUMBERS

OF REPORTS SUBMITTED PREVIOUSLY TO EPA

Page 32 of 46
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MRID 32539, 126621

BAYER AG 5600 Wuppertal 1

INSTITUT PUR TOXIKOLOGIE October 24, 1978

Report No.: 7882

t:<v,•.••-,

KEB 6447

LONG-TERM TOXICITY STUDY ON DOGS

(Two-year feeding study)

by

K. Hoffmann

and

P. Grb'ning

This Report may not be reproduced by any means, in whole

or in part. If required, further copies may be ordered

from the authors

Page 33 of 46
Bayer CropScience

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MRID 32541

BAYER AG Wuppertal-Elberfeld

INSTITUT Fl)R TOXIKOLOGIE April 12, 1979

Report No.: 8297

Confidential

Copyrtghl by Baye: A<3

Ihe use. L'liHE.y.iB:; o: diJtridulion

le only perm.llat/ with ir.o consent

Of Bayer AG

MEB 6447

MULTIGENERATION REPRODUCTION STUDY ON RATS

by

Dr. E. Lb'ser

Histopathological examinations conducted by

Consultox Laboratories Ltd., London

This Report may not be reproduced by any means, in whole

or in part. If required, further copies may be ordered

from the author

Page 34 of 46
Bayer CropScience

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MRID 48624

MEDIZINISCHE HOCHSCHULE HANNOVER

Abteilung flir experimentelle Pathologie

(Department of Experimental Pathology)

Leiter (Director): Prof. Dr. med. U. Mohr

BAYER AG

Institut fur Toxikologie Hanover

Fr.-Ebert-Str. 217 - 319 November 5, 1976

5600 Vuppertal-Blberfeld

Confr-: •• -if.!

Coj!V;h-: . ;-.i ; -.--si Af.

UK L'.-:,'. ;...,:v.:.;u., Of di»!;

Is oniy Msniiiiiud wi:ti tho

of Bayt-r AC.

I4EB 6447

Subchronic Toxicity Study on Rats

(Twelve-week feeding experiment)

Report No. R 640 a

R840a/MO-02-005634

Page 35 of 46
(*44 Bayer CropScience

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MRID 48625, 60226

BAYER AG Wuppertal-Elberfeld

INSTITUT FOR TOXIKOLOGIE December 16, 1974

Report No.: 5O71

Ihs •>*<<. iiAis.->>.:w or rS.'rthBtlon

Is only ne.-ir.!!'..5d wi'Ji !!>t c«awool

>.( Bnysr ACI.

MEB 6447

SUBCHRONIC TOXICITY STUDY ON DOGS

(Thirteen-week feeding experiment)

by

K. Hoffmann

and

G. Luckhaus

This report may not be reproduced by any means in whole or

in part. If required, further copies may be ordered from

the authors

Jill

5071 /MO-02-005453

Page 36 of 46
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MRID 89023

TERATOGENICITY TEST OF MEB 6447

IN PREGNANT RATS

March 13, 1981

Kesao Nagumo*, Yoshimi Teroki**, Tsugumichi Chiba

and Muneo Miyasoka

Laboratory of Embryology, the First Department of Anatomy,

St. Marianne University School of Medicine,

Kowosaki, Konagawa Prefecture, Japan

* Professor emeritus ** Assistant Professor

MO-02-010180 / MO-02-010180

Page 37 of 46
Bayer CropScience

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MRID 149336

\

A TERATOLOGICAL EVALUATION OF BAYLETON®

IN MATED FEMALE RATS

TOXICOLOGY REPORT NO. 324

by

Timothy M. Unger

Daniel Van Goethem

Thomas E. Shellenberger

MIDWEST RESEARCH INSTITUTE

425 Volker Boulevard

Kansas City, Missouri 64110

FINAL REPORT

August 31, 1982

MRI Project No. 7272-B

BC324/MO-02-005574

For

Mobay Chemical Corporation

Environmental Health Center

Stilvell, Kansas 66085

MIDWEST RESEARCH INSTITUTE '25 VOLKEP BOUUEVAHO. KANSAS CITY. MISSOURI 54110 • 816 753-7600

Page 38 of 46
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MRID 155075

BAYER AG

INSTITUTE OF TOXICOLOGY

Wuppertal-Elberfeld, 30.5.1984

Report no.: 12712

MEB.

(Triadimefon)

TWQ-GENERATIOH STUDY WITH RATS

(Supplementary study)

by

Dr. R. Eiben

Appendix;

Histopathologioal report by Life Science Research, England

Until they are published, the findings contained in this

report may only be used with the approval of BAYER AC.

Further reproduction of all or part of this report is not

permissible.

127121 MO 02-005612

Page 39 of 46
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MRID 40752101, 40865101

BAYER AG Hay 17, 1988

Fachbereich Toxikologie

Friedrich-Ebert-Str. 217-333

D-5600 Wuppertal 1

Federal Republic of Germany

Bayer Report No.: 16719 -

Co;-,.. ,— - »*•:.-.. ;'. •

 s««r '» IV ' ""SS"Of)0

MEB 6447

(Common Dame: Triadinefon, the Active Ingredient of BAYLETON)

CAKCIMOGENICITY STUDY ON NMRI MICE

(21-Month Administration in the Feed)

Report Section 1 of 3

by

Dr. E. Bomhard

and

Dr. S. Hahnemann

Study No.: T9017407

Study Completion Date: April 1986

Page 40 of 46
Bayer CropScience

MRID 41446201

Report MTD0149

Mobay 5o96

Date; March 22, 1990

US0046

Teratology Study in the Rabbit

with MEB 6447 (Triadimefon)

Authors

G.R. Clemens and R.E. Hartnagel Jr.

Data Requirement

U.S. EPA-FIFRA

Section 158.135

Guideline No. 83-3

Performing Laboratory

Toxicology Department

Miles Inc.

P.O. Box 40

Elkhart, IN 46515

Pages 1 to 155

RLE

509 6

MTD0149/MO-02-OOS963

-1- MTD0149

Page 41 of 46
HH Bayer CropScience

MRID 42089601

Supplemental Submission to EPA MRID 41446201

Study Title

Teratology Study in the Rabbit

with MEB 6447 (Triadimefon)

Report No.

HTD0149

Date

October 28, 1991

Data Requirement

U.S. EPA-FIFRA

Section 158.135

Guideline No. 83-3

Authors

G. R. Clemens and R. B. Hartnagel Jr.

Performing Laboratory

Toxicology Department

Miles Inc.

P.O. Box 40

Elkhart, IN 46515

Laboratory Project I.D.

Report No. 100026

US0046

MTD0149/MO-02-006369

Page 1 of 11

r*; r

Page 42 of 46
Bayer CropScience

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MRID 42153901

BAYER AG

Fachbereich Toxllcologie

Friedrich-Bbert-StraBse 217 - 333

D-5600 Wuppertal 1

Wuppertali 2^.10.

Report no. 2'Q"??*

MEB6447

Chronic Toxicity "and Cancerogenicity Studies on Wistar Rats

With Adiainigtration in Diet Over a Period of 10S Weeks

Part 1 of 3 parts

by

or. E. Bombard

and

veterinarian B. Schllde

Study No.: T 3027626

20774/M

Until they are published, the findings contained in this report

may only be used with the approval of BAYER AG, Further reproduction of all or part of this report ia prohibited.

Page 43 of 46
Bayer CropScience

US0046

MRID 43089301

BAYER AG

Fachbereich Toxikologie

Friedrlch-Ebert-Str. 217-333

42096 Wuppertal

Wuppertal, den 13.12.1993

Bericht-Nr.: 20774 A

MEB 6447

Chronic Toxicity and Cancerogenicity Study

on Wistar Rats with Administration in Diet

Over a Period of 105 Weeks

Addendum: The Incidence of Thyroid Follicular

Cell Tumors in wistar-Rats

- A Compilation of Historical Data -

Dr. E. Sander

and

B. Schilde

Study No.: T3027626

Prior to publication, the findings contained in this report may

only be used with the approval of BAYER AG. Further reproduction

of all or part of this report is not permitted.

Page 44 of 46
Bayer CropScience

MRID 47377101

BAYKR CROPSCIBNCE LP

07-1)72-1 L

STUDY TITLE

A Developmental Neurotoxicity Study with

Technical Grade Triadimefon in Wistar Rats

Data Requirement

U.S. EPA, OPPTS 870.6300

and OECD Test Guideline 426 (Draft)

Health Canada PMRA DACO No. 4.5.14

Authors

L.P. Sheets, Ph.D., R.G. Gilmore, B.S.

Author, Pathology Report

H.E. Hoss, DVM, M.S.

Study Completion Date

Report: February 20, 2008

Sponsor

Bayer AC

Bayer CropScience

Alfred Nobel Str. 50

40789 Monhcim

Germany

Test Facility

Bayer CropScience LP

Toxicology

17745 South Metcalf Avenue

Stilwell, Kansas 66085-9104

Study Number

07-D72-IL

Page I of 1179

M-297829-OM

US0046

Kcpnn Number

201766

Page 45 of 46
Bayer CropScience

MRID 47377101 supplement

HAY'lift CROKSCUiNCI: If

.VI UDY WO-HTMIUS0046

KH'OKTNI-MHHK

201766-1

Supplemental Submission To:

US EPA MRJD No. 47377101

STUDY TITLE

A Developmental Neuroto.vidty Study with

Technical Grade Triadimefon in Wistar Rats

Data Requirement

U.S. EPA, OPPTS 870.6300

and OECD Test Guideline 426 (Draft)

Health Canada PMRA DACO No. 4.5.14

Authors or Supplemental Kepurt

D. I... Van Coeihein and L. P. Sheets

Study Completion Date

Original Report: February 20, 2008

SuppliMiifniiil Repon: July 28, 2008

Sponsor

Bayer AG

Bayer CropScience.

Alfred Nobel Sir. M.I

40789 Monheini

Germany

Test Facility

Bayer CropScience L.P

Toxicology

17745 South Metculf Avenue

Stilwell, Kansas 66085-9104

Study .'Number

07-D72-II,

I «f 16

M-297829-02-1

Page 46 of 46