UNITED STATES ENVIRONMENTAL PROTECTION AGENCY WASHINGTON, D.C. 20460

OFFICE OF CHEMICAL SAFETY AND POLLUTION PREVENTION

MEMORANDUM

Date: June 29, 2015

SUBJECT: EDSP Weight of Evidence Conclusions on the Tier 1 Screening Assays for the
List 1 Chemicals

PC Code: See table, Attachment A Decision No.: NA Petition No.: NA
Risk Assess ment T y pe: NA
TXR No.: See table

, Attachment A
MRIDNo.:NA DP Barcode: NA Registration No.: NA Regulatory Action: NA
Case No.: NA
CAS No.: NA
40CFR: NA

FROM: Greg Akerman 

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Health Effects Divison

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Environmental

# Fat**~~ fec** **~~ ~n** **~** Office of Pesticide Program

THR O UG H: Je ss Rowland

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## ~~~ Deputy Divi s ion Director
Health Effects Division
Office of Pe s ticide Programs

TO: Jolene Trujillo Chemical Review Manager
Pe s ticide Re-Evaluation Division Office of Pesticide Programs Ver .A pr. 2010

EPA has completed its Weight of Evidence (WoE) assessment eval u ating results of the
Endocrine Screening Program (EDSP) Tier l screening assays for the List l chemicals.
The WoE documents for the 52 chemicals are li sted in Attac hm ent A a l ong with the chemical
and report identifiers

.

Page I of2Page 2 of 2 Attachment A. EDSP List 1 Chemicals

Chemical Name PC Code TXR Number 2,4-D 030001 0057151
Abamectin 122804 0057152 Acephate 103301 0057153
Acetone 044101 0057154
Atrazine 080803 0057155
Benfluralin 084301 0057156
Bifenthrin 128825 0057157 Captan 081301 0057158 Carbaryl 056801 0057159
Carbofuran 090601 0057160
Chlorothalonil 081901 0057161 Chlorpyrifos 059101 0057162 Cyfluthrin 128831 0057163 Cypermethrin 109702 0057164
DCPA 078701 0057165
Diazinon 057801 0057166
Dichlobenil 027401 0057167
Dimethoate 035001 0057168
EPTC 041401 0057169
Esfenvalerate 109303 0057170 Ethoprop 041101 0057171
Fenbutatin-Oxide 104601 0057172
Flutolanil 128975 0057173 Folpet 081601 0057174 Glyphosate 417300 0057175 Imidacloprid 129099 0057176 Iprodione 109801 0057177 Isophorone 847401 0057178
Linuron 035506 0057179
Malathion 057701 0057180 Metalaxyl 113501 0057181 Methomyl 090301 0057182
Metolachlor 108801 0057183
Metribuzin 101101 0057184
MGK-264 057001 0057185 Myclobutanil 128857 0057186
Norflurazon 105801 0057150 o-Phenylphenol 064103 0057146 Oxamyl 103801 0057142
PCNB 056502 0057138
Permethrin 109701 0057149
Phosmet 059201 0057145 Piperonyl Butoxide 067501 0057141
Pronamide 101701 0057137 Propargite 097601 0057148 Propiconazole 122101 0057144 Pyriproxyfen 129032 0057140
Simazine 080807 0057136
Tebuconazole 128997 0057143 Tetrachlorvinphos (TCVP) 083701 0057147
Triadimefon 109901 0057139
Trifluralin 036101 0057135

### EDSP: WEIGHT OF EVIDENCE ANALYSI S OF POTENTIAL INTERACTION WITH THE ESTROG EN, ANDROGEN OR THYROID PATHWAYS

CHEMICAL: IPRODIONE

O FFICE OF 

### P ESTICIDE 

### P ROGRAMS



### O FFICE OF 

### S CIENCE 

### C OORDINATION AND 

### P OLICY



### U.S E NVIRONMENTAL 

### P ROTECTION 

### A GENCYIPRODIONE FINAL

ii TABLE OF CONTENTS

Abbreviations...................................................................................................iii
Executive Summary..........................................................................................1

I. Introduction.......................................................................................4

II. Sources of Scientific Data and Technical Information......................................5

A. EDSP Tier 1 Screening Assays............................................................5

B. Other Scientifically Rele vant Information (OSRI)......................................5

III. Weight of Evidence (WoE) Evaluation.......................................................5

A. EDSP Tier 1 Screening Assays.............................................................8

B. Effects on Hypothalamic-Pituitary-Gonadal (HPG) Axis..............................9
1. Effects on Estrogen Pathway.....................................................9
2. Effects on Androgen Pathway..................................................14

C. Effects on Hypothalamic-Pituitary-Thyroidal (HPT) Axis...........................23

IV. Committee’s Assessment of Weight of Evidence..........................................25

A. Systemic/Overt Toxicity in the in vivo Tier 1 assays and OSRI .....................25

1. Tier 1 in vivo Assays.............................................................26

2. OSRI...............................................................................26

B. Estrogen Pathway...........................................................................28

C. Androgen Pathway.........................................................................28

D. Thyroid Pathway...........................................................................29

E. Conclusions.................................................................................29

V. EDSP Tier 2 Testing Recommendations....................................................29

VI. References.......................................................................................31

LIST OF TABLES: Table 1. Tier 1 Assay for Iprodione....................................................................8
Table 2. Estrogenic/Anti-Estr ogenic Pathway for Iprodione ....................................11
Table 3. Androgenic/Anti-Androge nic Pathway for Iprodione .................................18
Table 4. Thyroid Pathway for Iprodione ............................................................24

APPENDIX 1. EDSP Tier 1 Screening Assays..............................................................35
APPENDIX 2. Other Scientifically Relevant Information (OSRI)....................................42

APPENDIX 3. References Not Utilized in the Iprodione WoE Analysis.............................65IPRODIONE FINAL

iii Abbreviations

Abbreviation Terminology

A Androgen (hormonal pathway)

ADME Absorption, Distribution, Metabolism, Excretion

ALP Alkaline Phosphatase

ALT Alanine Aminotransferase

AMA Amphibian Metamorphosis Assay

ARTA Androgen Receptor Transcriptional Activation

AST Aspartate Aminotransferase

ANOVA Analysis of Variance
AOP Adverse Outcome Pathwa y A R Andro g en Rece p to r B max Bindin g at maximum BROD Benz y lox y resorufin-O-dealk y lase BUN Blood Urea Nitro g e n CA R Constitutive Androstane Rece p to r CFR Code of Federal Regulations

CG Cowper’s Gland

ChE Cholinesterase
ChEI Cholinesterase inhibition
CMC Carboxymethyl cellulose
CTA Comparative Thyroid Assay
CV Coefficient of Variation
CYP Cytochrome 450

DE R Data Evaluation Recor d DMSO Dimeth y l Sulfoxide DNA Deox y ribonucleic Aci d DO Dissolved Ox yg en DP Dorsolateral Prostrate
E Estro g en hormonal p athwa y EDSTAC Endocrine Disru p tor Screenin g and Testin g Advisor y Committee
EDRT Endocrine Disru p tor Review Team EDSP Endocrine Disru p tor Screenin g Pro g ram EE Ethin y l Estradiol ELISA Enz y me Linked Immunosorbent Assa y EOGRTS Extended One-Generation Re p roductive Toxicit y Stud y ( Rat )
E R Estro g en Rece p to r EROD Ethox y resorufin-O-dealk y lase ( or deeth y lase ) ERTA Estro g en Rece p tor Transcri p tional Activatio n EtOH Ethanol
F Female
F1 First filial g enerationIPRODIONE FINAL

iv Abbreviation Terminology

F2 Second filial g eneration
Fcd Fecundit y FIFRA Federal Insecticide, Fungicide and Rodenticide Act
FOB Field Observation Batter y F Q PA Food Q ualit y Protection Act
Frt Fertilit y FSH Follicle Stimulatin g Hormone FSTRA Fish Short-Term Re p roduction Assa y FT Flutamide GD Gestation Da y GGT Gamma- g lutam y l Trans p e p tidase GnRH Gonadotro p in-releasin g hormone GP Glans Penis GSI Gonado-Somatic Index
H Hi g h
HLL Hind Limb Len g th HPG H yp othalamic-Pituitar y -Gonadal Axis
HPLC/MS/MS High Pressure Liquid Chroma tography/Mass Spectroscopy

HPT H yp othalamic-Pituitar y -Th y roidal Axis
I Inade q uate IC50 Inhibitor y Concentration at 50% of res p onse

ICCVAM Interagency Coordinating Committee on the Validation of Alternative Methods

K d Equilibrium Dissociation Constant

K OW Octanol/Water Partition Coefficient

L Low dose LABC Levator Ani-Bulbocavernosus LAGDA Larval Am p hibian Growth and Develo p ment Assa y LC50 Lethal Concentration in 50% of test organisms

LD Lactation Day

LH Luteinizing hormone

LOAEC Lowest Observed Adverse Effect Concentration

LOAEL Lowest Observed Adverse Effect Level
LOQ Limit of Quantitation

M Male
MDL Minimum Detection Level MEOGRT Medaka Extended One Generation Re p roduction Test
MH Medium hi g h
ML Medium low
MoA Mode of Action MOE Mar g in of Ex p osure MRID Master Record IdentifierIPRODIONE FINAL

v Abbreviation Terminology

MROD Methoxyresorufin-O-dealkylase
MTC Maximum Tolerated Concentration
MTD Maximum Tolerated Dose
N Ne g ative
NE Not examined/evaluate d
NF sta g e Nieuwkoo p and Faber’s Sta g in g Atlas NOAEC No Observed Adverse Effect Concentratio n NOAEL No Observed Adverse Effect Level NS Not Statisticall y Si g nificant N R Not Re p orte d OCSPP Office of Chemical Safet y Pollution and Prevention
OECD Organization for Economic Co-Operation and Development

OPP Office of Pesticide Programs

ORD Office of Research and Develo p ment OSCP Office of Science Coordination and Polic y OSRI Other Scientificall y Relevant Informatio n P Positive
P Parental g eneratio n PC Positive Control
PC 10 Positive Control at 10% of response

PC 50 Positive Control at 50% of response

PND Post-Natal Da y POD Point of De p arture PPS Pre p utial Se p aration PROD Pentaox y resorufin-O-dealk y lase ( or de p ent y lase )
PX R Pre g nane X rece p to r QC Quality Control

RBA Relative Binding Affinity

RBC Red Blood Cells
RfD Reference Dose
RPC max Relative to Positive Control at maximum

SAP Scientific Advisor y Panel
SC Solvent Control

s.c Subcutaneous SDH Sorbitol deh y dro g enase SDWA Safe Drinking Water Act

SEP Standard Evaluation Procedure
SD Standard Deviation or Sprague-Dawley

SVL Snout-to-Vent Len g th SV Seminal Vesicles
T Th y roid ( hormonal p athwa y) T1WoERC EDSP Tie r 1 Wei g ht of Evidence Review Committee
T3 TriiodothyronineIPRODIONE FINAL

vi Abbreviation Terminology

T4 Thyroxine (tetraiodothyronine)

TP Testosterone Pro p ionate
T R Th y roid Rece p to r TSH Thyroid Stimulating Hormone

UDPGT Uridine Diphosphate Glucuronyltr ansferase (also known as UGT)

VC Vehicle Control
VO Vaginal Opening

VP Ventral Prostate VTG Vitello g eni n WoE Wei g ht-of-EvidenceIPRODIONE FINAL

Page 1 of 66 Executive Summary

The Endocrine Disruptor Screening Programs (EDSP) Tier 1 assay ba ttery is designed to provide the necessary empirical data to evaluate the pote ntial of chemicals to interact with the estrogen (E), androgen (A) or thyroid (T) signaling pa thways. This interacti on includes agonism and antagonism at the estrogen and androgen recep tors, altered steroidogenesis, as well as hypothalamic-pituitary-gonadal (HPG) and hypothalamic-pituita ry thyroid (HPT) axes. In
addition to the available Tier 1 assay data, other scientifically relevant information (OSRI), including general toxicity data and open literature studies of sufficient quality were considered in this weight of evidence (WoE) assessment.

In determining whether a pesticide chemical inte racts with E, A, or T hormone pathways, the number and type of effects i nduced, the magnitude and pattern of responses observed across studies, taxa, and sexes we re considered. Additi onally, the conditions un der which effects occur were considered, in part icular, whether or not endocrine-rela ted responses occurred at dose(s) that also resulted in general systemic toxicity or overt toxicity.

On March 5, 2014, the EDSP Tier 1 Assay Wei ght of Evidence Review Committee (T1WoERC) of the Office of Pesticide Programs (OPP) and the Office of Science Coordination and Policy (OSCP) conducted a weight-of-evidence (WoE) analys is of the potential in teraction of iprodione with the E, A or T hormone pathways. The T1 WoERC conclusions from the WoE evaluation in this report are presented by pathway (E, A and then T) beginning with the results of the Tier 1 in

vitro assays followed by in vivo mammalian and wildlife results, then the results of the cited
OSRI for mammalian and wildlife studies (40 CFR Part 158 and literature).

For the estrogen pathway, the Tier 1 in vitro aromatase, estrogen receptor (ER) binding, ER transactivation assay (ERTA) and the in vivo uterotrophic assays were negative. However, there appears to be a potential for interaction with th e estrogen pathway based on effects seen in the female pubertal assay [(delay in vaginal opening (VO), delay in age at first estrus and uterine histopathology) as well as OSRI studies (uterus atrophy and decrea sed number of corpora lutea). Decreases in fecundity/fer tility or reproductive/offspring parame ters were observed in fish shortterm reproduction assay (FSTRA) and birds (Par t 158 bobwhite quail study); however, it is noted that these effects have limited diagnostic utility for understanding a potential interaction with the estrogen pathway. Based on the evidence from male assays (OSRI), this potential interaction with the estrogen pathway may be due to al teration of the steroidogenesis pathway.

Evidence for potential interaction with the androg en pathway was from the result of the OSRI [( in vitro androgen receptor (AR) bindi ng/AR transactivation assay (ARTA) and steroidogenesis,
and in vivo Hershberger and the male pubertal assays )] and the FSTRA. These effects included decreases in the androgen-sensi tive organ weights in the Hershbe rger assay, delays in age andIPRODIONE FINAL

Page 2 of 66 weight at preputial separation (PPS) and decreased serum testosterone and androgen-sensitive organ weights in the male pubertal assay, as we ll as similar results in the Part 158 mammalian toxicity and special m echanistic studies.

Effects on male fish gonadal hi stopathology and effects on fertil ity were observed as well as reproductive effects in birds; again, it is noted th at effects on fecundity an d fertility have limited ability for informing whether iprodione in teracts with the androgen pathway. Although, iprodione did not demonstrate competitive bindin g to the AR, iprodione metabolites (produced in cell culture) showed evidence of direct interactio n with the AR. Additionally, there is evidence of iprodione potentially impacting steriodogenesis. Mechanistic st udies and studies published in the open literature suggest that iprodione may inte rfere with the active tran sport of cholesterol, the biosynthetic precurs or of testosterone, into Leydig cells, thereby resulting in potential decrease in overall steroid hormone levels, in cluding testosterone. Evidence for steroidogenesis inhibition is demonstrated by alterations in hor mone levels (testosterone, LH progesterone and aldosterone), decreases in androgen-sensitive tissu es weights, delayed PPS and increased adrenal gland weights in mammals.

There is no convincing evidence of an interac tion with the thyroid pa thway. In the female pubertal assay, serum T 4 concentrations were reduced; howev er, there were no effects on thyroid stimulating hormone (TSH), thyr oid weight or histopathology. Additionally, no thyroid-related
effects were observed in the Pa rt 158 mammalian toxicity studies. No thyroid-related effects were noted in the amphibian metamorphosis assay (AMA).

Overall, based on the weight of the evidence there appears to be a potential for iprodione to alter steriodogenesis, which may affect the estrogen and androgen pathways in mammals and wildlife.

For iprodione a point of departure (POD) of 0.5 mg/kg/day is used for human health risk assessment. The study by Blystone et al. (2007) was accepted to satisfy the requirement for the male pubertal assay. In that study there was a dose-dependent decr ease in testosterone levels in males at 50 mg/kg/day, the lowest obs erved effect level (LOAEL). Therefore, a 10X Uncertainty Factor (UF L ) was applied ( i.e ., for the use of a LOAEL) resulting in an extrapolated NOAEL of 0.5 mg/kg/day ( i.e , the POD). This POD is used for establishing the chronic Reference Dose (RfD) and to calculate the Margins of Exposure (MOEs) for dietary and non-dietary exposure risk assessments, re spectively. The POD is approximately 300-fold lower than the lowest dose (150 mg/kg/day) that caused estrogen-related effects in the female pubertal assay. Consequently, the ch ronic RfD and the MOEs are likely to be protective of the
endocrine-related effects s een in the pubertal assay.

Therefore, mammalian EDSP Tier 2 testing is no t recommended for iprodione since additional testing is not expected to impact EPA’s current regulatory point of departures and endpoints for human health risk assessments.IPRODIONE FINAL

Page 3 of 66 For wildlife, the current chronic fish NOAEC value is 0.26 mg/L based on a fathead minnow early life-cycle study (LOAEC is 0.55 mg/L base d on survival). Effects on fecundity were
observed in the FSTRA at ≥ 0.084 mg/L. In addition, effects on fertility were observed at 0.60 mg/L and potential treatment-re lated effects on male gonadal hi stopathology were observed at 0.084 mg/L. Therefore, the EDSP Tier 2 Medaka Extended One Generation Reproduction Test (MEGORT) is recommended.

For birds, Part 158 avian reproduction studies w ith both northern bobwhite quail and mallard duck are available for iprodione. In both spec ies, effects on reproductive parameters were observed. The type of data obtained from Pa rt 158 avian reproduction studies (OCSPP 850.2300) are considered sufficient for evaluating potential reproductive effects to birds from iprodione exposure. Relevant to EDSP, additional testing is not recommended.IPRODIONE FINAL

Page 4 of 66 I. Introduction

The Endocrine Disruptor Screening Programs (EDSP) Tier 1 assay ba ttery is designed to provide the necessary empirical data to evaluate the pote ntial of chemicals to interact with the estrogen (E), androgen (A) or thyroid (T) signaling pa thways. This interacti on includes agonism and antagonism at the estrogen and androgen recep tors, altered steroidogenesis, as well as hypothalamic-pituitary-gonadal (HPG) and hypothalamic-pituita ry thyroid (HPT) axes. In
addition to the available Tier 1 assay data, other scientifically relevant information (OSRI, including general toxicity data and open literature studies of sufficient quality were considered in this weight of evidence (WoE) assessment.

In determining whether a pestic ide chemical interacts with E, A or T hormone pathways, the number and type of effects i nduced, the magnitude and pattern of responses observed across studies, taxa and sexes were considered. Additionally, the conditions under which effects occur were considered, in part icular, whether or not endocrine-rela ted responses occurred at dose(s) that also resulted in general systemic toxicity or overt toxicity.

On March 5, 2014, the EDSP Tier 1 Assay Wei ght of Evidence Review Committee (T1WoERC) of the Office of Pesticide Programs (OPP) and the Office of Science Coordination and Policy (OSCP) conducted a weight-of-evidence (WoE) analys is of the potential in teraction of iprodione with the E, A or T hormone pathways. The T1 WoERC conclusions from the WoE evaluation in this report are presented by pathway (E, A and then T) beginning with the results of the Tier 1 in

vitro assays followed by in vivo mammalian and wildlife results, then the results of the cited
OSRI for mammalian and wildlife studies (40 CFR Part 158 and literature).

Iprodione has an Log octanol water partition coe fficient of 3.1, and bioconcentration factors for bluegill sunfish are 51–116 with half -live depuration rates of <1 day. Iprodione is not expected to volatilize substantially as its vapor pressure is relatively low (2.7 x 10 -7 torr). The major routes of dissipation for iprodione in the environm ent are by hydrolysis in neutral and alkaline environments (half-lives: pH 7 = 4.7 days; pH 9 = 27 minutes) and microbial degradation under
both aerobic and anaerobic conditions.

The available information considered in determin ing the potential interact ion of iprodione with the E, A and/or T pathways include submitted EDSP Tier 1 assays and/or other scientifically relevant information (OSRI) such as general toxi city studies and other pu blished articles. These data are summarized in Sections III.A through I II.C. An analysis of the data submitted to the Agency, using the WoE approach outlined by th e Agency (USEPA, 2011), is presented in
Section IV. The EDSP Tier 2 studies recommendations are presented in Section V.IPRODIONE FINAL

Page 5 of 66 II. Sources of Scientific Data and Technical Information

A. EDSP Tier 1 Screening Assays

The Tier 1 assays and/or other scientifically relevant information (OSRI) submitted to satisfy the agency’s test order are shown below in Table 1. Executive Summaries are presented in Appendix

1.

B. Other Scientifically Relevant Information (OSRI)

In response to the Agency’s Test Orders, data believe d to be relevant to one or more of the Tier 1 assays were submitted as OSRI by the Test Orde r recipients and/or the public. This included studies published in the open literature and/or data submitted to support pesticide registration ( e.g. , Part 158 guideline studies). The Agency’s re view of the initial OSRI is provided in the Report of the Endocrine Disruptor Review T eam on iprodione (USEPA, 2010). Since then, the Agency has also conducted a more recent search of the available scientif ic literature for any
additional relevant information. Su mmaries of the Tier 1 assays and the studies cited as other scientifically relevant information (OSRI) ar e presented in Appendix 1 and 2, respectively. Additionally, literature/studies c onsidered but not util ized for the WoE analysis are listed in Appendix 3.

III. Weight of Evidence (WoE) Evaluation

The principles, criteria and approach used in the WoE determination on the potential of a
substance to interact with endoc rine-related processes (i.e., E, A, or T hormone pathways) were as described in the WoE guidance document (USEPA, 2011) and presen ted at the 2013 FIFRA Scientific Advisory Panel (SAP) (USEPA, 2013). The weight of evidence process identifies how
the individual lines of evidence are asse mbled and integrated along two concepts ( i.e

., complementarity and redundancy) within the co nceptual framework of an adverse outcome pathway). Broadly, there are four main st eps outlined in the guidance which provide the foundation for WoE evaluations. The first step is to evaluate the indivi dual studies for their scientific quality and relevance in evaluating potential endocrine interaction(s). The second step is to integrate the data along different levels of biological organization while examining the extent of complementarity ( i.e ., the concordance of endpoints within an assay that measures multiple endpoints) and redundancy ( i.e ., the concordance of endpoint s/responses across assays) in the observed responses across these different le vels of biological orga nization. The third step
is to characterize the main lines of evidence as well any conclusions. Finally, the last step is to
evaluate whether additional te sting is needed based on the evidence and conclusions described
above.IPRODIONE FINAL

Page 6 of 66 As mentioned, the first step is to assemble and evaluate the available scientific data. Data for the
EDSP Tier 1 WoE evaluation falls into one of two categories: 1) EDSP Tier 1 data, or 2) other scientifically relevant information (OSRI). The EDSP Tier 1 data include a battery of 11 assays consisting of in vitro and mammalian and wildlife in vivo assays. The Tier 1 assays were designed specifically to evaluate a number of key biological events including potential effects on receptor binding (estrogen and androgen agonist and antagonist), steroidogenesis, and other effects on the HPG and HPT axes. OSRI may in clude published literature studies as well as studies conducted under USEPA (o ften referred to as Part 158 data) or OECD guidelines submitted in support of pesticide registrations. E ach study is evaluated for scientific quality and relevance for informing interactions with the E, A, or T pathway. Additionally, the consistency of the responses in the individual study is evaluated. For the Tier 1 in vivo assays, often multiple endpoints are measured in each assay.

Evaluation of the potential confounding effects of overt toxicity in the study, as well as the relative degree of diagnostic utility of a speci fic endpoint for discerni ng whether or not the chemical has interacted with the endocrine syst em, are considered. The co llective response of the individual endpoints, as well as the conditions under which they were expressed, are considered when evaluating an overall indi cation of potential interaction as measured by the study.

The second step in this WoE process is to form ulate hypotheses and integr ate the available data along different levels of biological organization. Two keys elements in the integration of data, as well as characterizing the extent to which the available data suppor t a hypothesis that a chemical has the potential to interact E, A, or T path ways are the concepts of complementarity and redundancy. These two concepts provide a basis for considering the plau sibility, coherence, strength, and consistency of the body of evidence. The current EDSP Tier 1 screening assays are
meant to evaluate whether or not a chemical can in teract with E, A, and T consisting of different levels of biological organization from a molecular initiating ev ent, such as receptor binding, through potential adverse effects in apical endpoint s such as sexual develo pment and fecundity at the whole organism level. The extent of expressi on of responses to high er levels of biological organization can indirectly provide information on potential compensatory capabilities of an individual organism.

After the data have been assembled and integrated, the third step is to characterize the main lines of evidence along with the conclusions; this characterization involves three components. The first component is whether the data provide releva nt, robust and consistent evidence in terms of complementarity and redundancy, as well as biologi cal plausibility. Second, is at what level of biological organization were the responses observed and whether organisms exhibit compensatory responses at higher levels of biological organization? Finally, under what conditions did the responses occur including co nsideration of whether the responses were observed in the presence of overt or systemic toxicity? The presence of overt and/or systemicIPRODIONE FINAL

Page 7 of 66 toxicity introduces uncertainty in the ability to distinguish eff ects specifically related to an
endocrine-related effect from a non-endocrine toxic response.

This uncertainty in distinguishi ng whether the responses were endoc rine-related was discussed at the FIFRA SAP meeting in July 2013 that evaluated scie ntific issues associ ated with the WoE evaluation of the EDSP Tier 1 screening proce ss. In October 2013, the SAP stated that , “ In summary, the Panel agreed that little, if any, we ight should be placed on signs of endocrine disruption in the presence of overt toxicity. All effects in endocrine se nsitive tissues should be evaluated in terms of primary interactions with the endocrine system vs. secondary effects related to toxicity in non-endocrine organs or overall disruptions in homeostasis” (USEPA, 2013).

For these WoE analyses, overt toxicity was gene rally defined in accordance with EPA’s current approach as used by OPP in reviewing 40 CFR Part 158 studies for both human and ecological risk assessments. Specifically, in these analyses, the effects that EPA considered to be potential evidence of overt toxicity include d, but were not limited to: mortal ity; clinical signs such as tremors, ataxia and abnormal swimming (fish and aquatic-phase amphibians); and body weight
decreases of ≥ 10% in mammals. Additionally, other effects including morphological ( e.g., organ weights/histopathology), biochemical ( e.g., blood chemistry), and other clinical signs ( e.g. , lethargy) were also considered when evaluating overt toxicity, es pecially if the effects were extreme. In some instances, one parameter ( i.e ., death or >10% decrease in mammalian body weight) was sufficient to consider a dose/concentra tion to be overtly toxic. However, in other instances, more than one parameter was needed to determine overt toxicity. For example, in the FSTRA, generally, body weight decr eases were considered along with other responses when assessing potential overt toxicity. Additionally, effects which were considered to be signs of systemic toxicity were also captured and these ef fects were generally cons idered as less severe forms of toxicity ( e.g., changes in organ weights or blood chemistry). The circumstances for
which a dose/concentration was co nsidered overtly toxic for a pa rticular study are described in
Section IV.A.

In summary, EPA considers multiple lines of evidence in including the observed responses in the Tier 1 assays and OSRI in the context of a chemical’s physical/chemi cal properties and its
known modes of action in its overal l characterization of a chemical ’s potential to interact with the E, A or T pathway. Adequately addressing the three main components described above is fundamental to the WoE process and in determ ining whether additional data are needed. In addition to characterizing the WoE, reviewers also considered: 1) uncertainties and their potential impact to conclusions; 2) discussion of key studies; 3) description of inconsistent or conflicting data; 4) overall strength of eviden ce supporting a conclusion; and, 5) what, if any, additional data are needed and why. Assessing th e need for additional data is based on a caseby-case analysis which took all availa ble toxicity data into account.IPRODIONE FINAL

Page 8 of 66 The WoE approach involved consideration of data (i.e., lines of evidence) from the EDSP Tier 1 assays and OSRI which are depicted in Tables 2 - 4

. These tables contain data that are considered scientifically and biol ogically relevant with regard to a treatment -related effect which supports a conclusion of whether a substance has the potential to interact with the E, A, or T pathway. Effects that occurred in the presence of overt toxicity are discussed in the text for each respective pathway (E, A or T) but are not reported in the table for E, A or T.

A. EDSP Tier 1 Screening Assays

The Tier 1 assays submitted in response to the agency’s test order for iprodione are shown below
in Table 1

.

Table 1: Tier 1 Screening Assays for Iprodione.
Tier 1 Assays Test Guideline Test Order Status

ER Binding Assay (Rat uterine cytosol) OCSPP 890.1250 Requirement Satisfied (MRID No. 48617202)

ER α Transcriptional Activation Assay (Human cell line HeLa 9903) OCSPP 890.1300; OECD 455 Requirement Satisfied (MRID No. 48617203)

AR Binding Assay (Rat prostate cytosol) OCSPP 890.1150 Requirement Satisfied by OSRI (Blystone et al ., 2007 and 2009; Fail et al ., 1994)

Steroidogenesis Assay OCSPP 890.1550; OECD 456 Requirement Satisfied by OSRI (Blystone et. al. 2007)

Aromatase Assay (Human recombinant microsomes) OCSPP 890.1200 Requirement Satisfied (MRID No. 48617201)

Uterotrophic Assay (Rat) OCSPP 890.1600; OECD 440 Requirement Satisfied (MRID No. 48617205)

Hershberger Assay (Rat) OCSPP 890.1400; OECD 441 Requirement Satisfied by OSRI (Blystone et al ., 2009)

Pubertal Female Assay (Rat) OCSPP 890.1450 Requirement Satisfied (MRID No. 48671502)

Pubertal Male Assay (Rat) OCSPP 890.1500 Requirement satisfied by OSRI (Blystone et. al. 2007)

Fish Short-term Reproduction Assay OCSPP 890.1350; OECD 229 Requirement Satisfied (MRID No. 48617204)

Amphibian Metamorphosis Assay (Frog) OCSPP 890.1100; OECD 231 Requirement Satisfied (MRID No. 48671501)IPRODIONE FINAL

Page 9 of 66 B. Effects on Hypothalamic-Pit uitary-Gonadal (HPG) Axis

1. Effects on Estrogen Pathway

The potential for iprodione to interact with the estrogen pathway is summarized in Table 2. The table depicts the lines of evidence indicating pot ential interaction with the pathway. The various targets of the estrogen pathway across the releva nt Tier 1 assays are delineated to facilitate determination of potential for estrogenic, anti-estr ogenic, or HPG axis effects. This table also includes HPG-relevant findings from data evaluated as OSRI. Effects that occurred in the presence of overt toxicity are discussed in the text but are not reporte d in the table and not considered further in the WOE assessment.

Iprodione was negative (or not interactive) in the in vitro ER binding, ERTA, and aromatase assays. Iprodione was also negati ve in an OSRI ERTA (Kojima et al ., 2004) in Chinese hamster ovary (CHO) cells. Iprodione di d not induce changes in uter ine weight in the Tier 1 in vivo uterotrophic assay.

The test order for steroidogenesis as say was satisfied by OSRI (Blystone et al., 2007; MRID No.48279201). The authors examined ipr odione effects on steroidogenesis in vitro in testicular sections from male rats prev iously exposed to iprodione in vivo . Although estradiol
concentrations were not measured in this study, there was an effect of iprodione on steroidogenesis based on the observe d dose-dependent decreases in testosterone concentrations, and decreasing trends (not st atistically significant, NS) in the concentrations of 17 α -hydroxy progesterone and progesterone. Th e authors noted that the observed decreases suggest that iprodione affects steroidogenesi s at CYP11A (P450scc; cholestero l side-chain cleavage enzyme), StAR (steroidogenic acute regulat ory protein), or a preceding even t. Disruption of these events could affect production of all steroid hormone s, including estrogens.

In the female pubertal assay there were no si gnificant treatm ent-related effects on estrous cyclicity parameters, or in ovari an or uterine weights. A delay in the age at VO (3.9 days), and associated delay in the onset of first estrus (3.5 days), was obse rved at 300 mg/kg/day, in the absence of overt toxicity. Increa se in body weight at VO (11%) and decrease in pituitary weight (12%) were also observed at 300 mg/kg/day. Dose-d ependent increased incidence of minimal to mild decrease uterine horn diameter with associated decreased stromal development was observed at 150 mg/kg/day (2/16 females) and at 300 mg/kg/day (5/16 females) compared to controls (0/16). While no overt toxicity was observed in th e female pubertal assay, dosedependent increases over control were observed in liver weight (8 to 14%) and adrenal gland weight (24% to 42%) at 150 and 30 0 m g/kg/day, without clinical chemistry or histopathological
correlates that would indicate adverse liver ch anges; adrenal glands were not examined histopathologically.IPRODIONE FINAL

Page 10 of 66 In the FSTRA, decreased fecundity of 45 a nd 52% was observed at the intermediate (0.084 mg a.i./L) and high concentrations (0.60 mg a.i./L), respectively, p<0.05) and a significant (p<0.05) decrease in fertility of 12% was observed at the high concentration, in the absence of overt toxicity. There were no significant changes in gonada l histopathology and plasma vitellogenin (VTG) concentration.

In the mammalian Part 158 studies , estrogen-related effects were observed; however, in most
instances the effects co-occurred with overt toxi city or severe systemic toxicity. In the rat subchronic study, increased incidence of the follo wing observations were made at the top two doses (266 and 408 mg/kg/day), which also s howed overt toxicity (30%–50% decrease body weight): thin uterine horns (gross observation, 5/10 and 10/10), uterus atrophy (2/10 and 9/10) and decreased number of corpora lutea in the ova ries (2/10 and 4/10). In the same study, relative uterus weights were decreased 26% at 408 mg/kg/day compared to controls. In the mouse subchronic study, increased incidence of thin uterine horns (gross observation, 4/10), uterus atrophy (4/10) and no cor pora lutea in the ovaries (2/10), along with a decrease (29%) in absolute uterus weights were observed at 1303 mg/kg/day and coin cided with high incidence of swollen abdomen (7/10) and a 98% increase in ad justed liver weight. Higher incidences/percent changes of the same reproductive and liver e ffects were observed at a higher dose (2608 mg/kg/day) but co-occurred w ith clinical signs suggestive of overt toxicity (lethargy, piloerection). In the mouse carcinogenicity study, uterine weights (adj usted weights) were decreased by 69% at 100 weeks. Fewer high-dose females (16/50) displayed uteri that were thickened compared to control (39/50). Absolute and relative ovarian weights were decreased at 52 weeks at all doses, but there was no dose respon se. At 100 weeks, absolute and relative ovary weights were decreased by 53% (N S) at the high dose. At the hi gh dose, luteinization of the interstitial cells of the ovary was seen at 52 weeks (8/13, versus 0/13 in control) and at 100 weeks (23/50 versus 6/50 in control). In addition, the incidence of luteoma of the ovaries was increased at the high dose compared to control.

In Part 158 avian reproduction studies, there wa s a reduction in duck hatchling survival at Day 14 (p<0.05 compared to control) and a re duction in quail hatchl ing body weight (p<0.05, 6.0 g vs. 6.3 g control) and number of eggs laid (524 vs. 692 in the control) at the high dose (1000 ppm). However, the reduction in mallard reproduction was observed along with 5 mallards exhibiting loss of coordination or wing droop. In the Part 158 fish early-life cycle study, survival and/or growth was affected at ≥ 0.55 mg/L.IPRODIONEFINALPage 11 of 66Table 2. Estrogenic/Anti-Estrogenic Pathway for IprodioneLines of Evidence Indicating Potential Interaction withthe Estrogenic/Anti-Estrogenic Pathway for Iprodione1Study Type / LiteratureCitation ER Binding ER Activation Steroidogenesis Sex Steroid Hormone Uterine Weight/Histology Ovarian Weight/GSI Ovarian/Gonad Staging and Histopathology Pituitary Weight Estrous Cyclicity Age & Weight at VO 2° Sex Characteristics Fertility(Frt) /Fecundity (Fcd)/ Reproductive success (Repro) Vitellogenin (VTG) Systemic Toxicity Observed 2 Overt Toxicity Observed 3EDSP Tier 1 AssayER Binding(MRID 48617202)NERTA(MRID 48617203)NAromatase(MRID 48617201)NSteroidogenesisRequirement satisfied by OSRI (Blystoneet al.2007; MRID No.48279201; see below)Uterotrophic(MRID 48617205)NX(H)Female Pubertal Rat(MRID 48671502)P4NN↓12%(H)Age↑3.5days(H)Age,↑3.9days;BW,↑11%;(H)↑LW(M, H)↑AW(M, H)NFSTRA(MRID 48617204)NENNNFrt:↓12%(H);Fcd:↓45,52% (M, H)NNOSRIERTA(Kojimaet al., 2004)NIPRODIONEFINALPage 12 of 66Table 2. Estrogenic/Anti-Estrogenic Pathway for IprodioneLines of Evidence Indicating Potential Interaction withthe Estrogenic/Anti-Estrogenic Pathway for Iprodione1Study Type / LiteratureCitation ER Binding ER Activation Steroidogenesis Sex Steroid Hormone Uterine Weight/Histology Ovarian Weight/GSI Ovarian/Gonad Staging and Histopathology Pituitary Weight Estrous Cyclicity Age & Weight at VO 2° Sex Characteristics Fertility(Frt) /Fecundity (Fcd)/ Reproductive success (Repro) Vitellogenin (VTG) Systemic Toxicity Observed 2 Overt Toxicity Observed 3Steroidogenesis(Blystoneet al.2007;MRID 48279201)NE5Subchronic oral toxicity(Rat; MRID 42960701)NNNNX(M,H)X(M, H)Subchronic oral toxicity(Mouse;MRID 48125502)NNNNX(ML,M,MH,H)X(MH,H)Chronic oral toxicity(Dog; MRIDs 00144391,41327001 and42211101)NNNNX(M,H)NChronic oral toxicity /carcinogenicity(Rat; MRID 42787001)NNNNX(M,H)NCarcinogenicity(Mouse;MRID 42825002)P6(H)NP7(H)NX(H)NDevelopmental toxicity(Rat; MRIDs 00162984,40514901)NENENENENNE NDevelopmental toxicity(Rabbit;MRID 00155469)NNENENENNEX(H)IPRODIONEFINALPage 13 of 66Table 2. Estrogenic/Anti-Estrogenic Pathway for IprodioneLines of Evidence Indicating Potential Interaction withthe Estrogenic/Anti-Estrogenic Pathway for Iprodione1Study Type / LiteratureCitation ER Binding ER Activation Steroidogenesis Sex Steroid Hormone Uterine Weight/Histology Ovarian Weight/GSI Ovarian/Gonad Staging and Histopathology Pituitary Weight Estrous Cyclicity Age & Weight at VO 2° Sex Characteristics Fertility(Frt) /Fecundity (Fcd)/ Reproductive success (Repro) Vitellogenin (VTG) Systemic Toxicity Observed 2 Overt Toxicity Observed 3Two-generationreproduction(Rat; MRID 41871601)NENENNENX(M,H)X(M, H)Avian reproduction(Duck;MRID 00086840)NNX(H)Avian reproduction(Quail;MRID 00099126)P8(H)N N1. Key to responses: L=Low treatment, ML=Medium-low treatment, M=Medium treatment, MH=Medium-high treatment, H=High treatment.Arrows (↓or↑) indicate the direction of the response. Ashaded cell indicates that is parameter is not routinely evaluated or is not applicable in this assay.Changes in weight are absolute unless otherwise indicated. LW= Liver weight; AW= Adrenal weight2. The systemic toxicity in the Tier 1 assays are presented in this column (e.g. KW= kidney weight). The systemic toxicity for the OSRI is indicated by an X inthis column. For details see Section IV. A3. The overt toxicity in the Tier 1assays are presentedin this column (e.g.↓BW). The overt toxicity for the OSRI is indicated by an X in this column. Fordetails see Section IV. A4. Dose-dependent increased incidence of minimal to mild decrease uterine horn diameter with associated decreased stromal development was observed at 150mg/kg/day (2/16 females) and at 300 mg/kg/day (5/16 females) compared to controls (0/16).5. The effects of iprodione on steroidogenesis was evaluated in the OSRI (Blystone et al. 2007; MRID 48279201); however, estradiol production was notevaluated in this study.6. Uterine weights (adjusted weights) were decreased by 69% at 100 weeks. Fewer high-dose females (16/50) displayed uteri thatwere thickened compared tocontrol (39/50).7. At the high dose, luteinization of the interstitial cells of theovary was seen at 52 weeks (8/13, versus 0/13 in control) and at 100 weeks (23/50 versus 6/50 incontrol). In addition, the incidence of luteoma of the ovaries was increased at the high dose compared to control.8. A reduction in quail hatchling body weight (p<0.05, 6.0 g vs. 6.3 g control) and number of eggs laid (524 vs. 692 in the control) at the high dose (1000 ppm).P Positive findingsN Negative findings (in vitro) / No effect (in vivo)NE Not examinedIPRODIONE FINAL

Page 14 of 66 2. Effects on Androgen Pathway

The potential for iprodione to interact w ith the androgen pathway is summarized in Table 3. The table depicts the lines of evidence indicating pot ential interaction with the pathway. The various targets of the androgen pathway across the relevant Tier 1 assays are delineated to facilitate determination of potential for andr ogenic, anti-androgenic, or HPG axis effects. This table also includes HPG-relevant findings from data evaluated as OSRI. Effects that occurred in the presence of overt toxicity are discussed in the text but are not reporte d in the table and not considered further in the WOE assessment.

The requirements for the in vitro AR binding and steroidogenesis assays were satisfied by OSRI studies (Blystone et al . 2009 and 2007; Fail et al. 1994). In an AR binding assay similar to the Tier 1 assay (Fail et al. (1994), iprodione did not bind to the AR; however a metabolite of iprodione bound 50% of the available receptor at approximately 79 μM. In another study by Blystone et al. 2009 iprodione competitively bound to th e AR in a whole cell system using
human AR with an IC 50 of 86.0 μ M. Blystone et al. (2009) also investigat ed AR transcriptional activation (ARTA) in absence (androgenic assay) and presence (anti-andr ogenic assay) of the strong androgen dihydrotestosterone . Iprodione was negative in the androgenic ARTA assay up to the limit of cytotoxicity, but was positiv e for anti-androgenic activity at 30 and 100 μM. However, iprodione was negative for AR agonism and antagonism in an ARTA considered as OSRI that was conducted in CHO cells (Kojima et al ., 2004). In an ex vivo steroidogenesis assay (Blystone et al., 2007), testosterone production was decreas ed (p<0.05) at ip rodione doses of

≥ 100 mg/kg/day and progesterone levels were wa s decreased (p<0.05) at an iprodione dose of 200 mg/kg/day.

The requirements for the in vivo Hershberger, and male pubertal assays were satisfied by OSRI studies. In a Hershberger assay testin g only anti-androgenic activity (Blystone et al., 2009), organ weight decreases (compared to control) we re observed in the ventra l prostate (30% at 200 mg/kg/day) and levator ani-bu lbocavernosus (LABC) muscle (10%, 11% and 21% at 50, 100 and 200 mg/kg/day, respectively) in the absence of ove rt toxicity; weights of other male accessory tissues did not change with treatment. At 200 mg/kg/day, these effects co-occurred with increased liver weights (10%) and adrenal gland weights (44%); liv er weight was also increased at 100 mg/kg/day but not at 50 mg/kg/day. In the male pubertal assay (Blystone et al ., 2007), seminal vesicle weight decreased 24% and ep ididymides weight decreased 11% at 200 mg/kg/day, and a dose-dependent delay in age at preputial separation (PPS) was observed (2.0 days at 100 mg/kg/day and 2.8 days at 200 mg/kg/day; p ≤ 0.5) accompanied by a dosedependent increase in body weight at PPS (9-16% of control), in the absence of overt toxicity. Also in the pubertal assay, serum testosterone levels were signifi cantly decreased in all treatment groups in a dose-dependent manner (73%, 83% and 89% of control at 50, 100 and 200 mg/kg/day, respectively). These effects co-occurred with increased liver we ight (14%), increased adrenal gland weight (40%), and decrease serum aldos terone levels (13%).IPRODIONE FINAL

Page 15 of 66 In the FSTRA, there was an increase in inters titial cell hyperplasia a nd asynchronous gonadal development in male fish. While interstitial hyp erplasia was not observed in any of the control males, this lesion was present in males from a ll the iprodione exposed groups: 2 of 7 (29%) dose group 3 males; 3 of 7 (43%) of dose group 2 male s; and 1 of 5 (20%) of dose group 1 males. However, severity of the interstitial hyperplasi a was minimal to mild in all cases and there was
not a clear increase in incide nce or severity with increa sed dose concentration. Mild asynchronous gonad development was observed in 25% of the males in the control compared to
mild incidences in 40–43% of the fish in the iprodione-treated gr oups. At the high dose, fertility was significantly reduced by 12%. However there were no changes in other endpoints such as male secondary sex characteristics, GSI, or male tubercle score.

A variety of effects suggesti ng interaction with the androge n pathway were observed across species in mammalian Part 158 studi es, however in most instances the effects co-occurred with overt toxicity or systemic t oxicity. A common finding was incr eased liver and adrenal gland weights at the highest dose tested, and occasionall y in lower dose groups as well. Serum levels of
testosterone were not determined in any of the Part 158 studies.

In the subchronic oral study in rats (M RID 42960701), the following were observed at 355 mg/kg/day and co-occurred with severe overt to xicity (61% decrease in body weight and early group termination for humane reasons): 4/10 male s had small testicles (o bserved grossly) and
6/10 were found to have seminiferous tubule atrophy; 6/10 had abnormal spermatozoa and 5/10 had spermatozoa absent; 9/10 had prostate atrop hy; 7/10 had reduced seminal vesicle secretion. Prostate atrophy was also observed at 252 mg/kg/ day in 5/10 males but it also co-occurred with overt toxicity (45% decrease body weight and clinical signs). In the same subchronic rat study, males also displayed increased incidence of a variety of adrenal hist opathological changes and decreased pituitary weights (20-35 % of control) at doses that coincided with overt toxicity.

Most of the testicular, prostate and adrenal effects observed in th e subchronic rat assay were also observed in the chronic/carcinogeni city rat study but at a lower dos e (69 mg/kg/day) and in the absence of overt toxicity but in the presence of liver toxicity (MRID 42637801; 42787001). In addition, increased incidence of testicular interstitial tumors was observed at 69 mg/kg/day (29/60 rats compared to 3/60 in controls). In the mouse carcinogenicity study (MRID 42825002),
testicular effects observed incl uded increased number of gro ss observations at 604 mg/kg/day (masses in testis, small and/or flaccid testis ), and microscopic obser vations (generalized vacuolation and hypertrophy of interstitial cells) at ≥ 115 mg/kg/day; these effects co-occurred with liver toxicity. In the chronic dog study (MRID 41327001), males had decreased prostate weights (22–41%) at ≥ 24.9mg/kg/day in the abse nce of overt toxicity but co-occurring with liver toxicity and increased adrenal weight s at the high dose only (145 mg/kg/day).IPRODIONE FINAL

Page 16 of 66 In addition to the required Part 158 studies, a series of special mechanistic studies were conducted with iprodione to better delineate the potential for disruption of the androgen pathway. In a special developmental toxicity study (R epetto-Larsey, 1997), anogenital distance (AGD) was decreased (an indication of anti-androgenic effects) in male pups at 120 and 250 mg/kg/day; overt toxicity (maternal deaths) was observed only at 250 mg/kg/day. Another developmental study by Wolf et al. (1999) reported no changes in AGD or other androgen-related parameters associated with demasculinization at 100 mg/kg/day iprodione (only dose tested).

Three mechanistic studies were conducted to examine iprodione-rela ted effects on hormone plasma levels in male rats. Two studies (Chuzel, 1998, 2001) examined the time course of
testosterone and luteiniz ing hormone (LH) plasma levels afte r administration of a single dose of iprodione. These studies demonstrated early d ecreases in testosterone concentrations with subsequent increases in LH concentrati ons. A third study (Kennels, 2002b) examined testosterone and LH plasma levels for 24-hours post-dose after 14 days of iprodione treatment; the results were similar to the single-dose studi es. In all three studies, hormone plasma levels were generally similar to cont rol at the 24-hour post-dose.

Another two studies examined effects on testoste rone production in male rats after iprodione treatment and human chorionic gonadotrophin (hCG) challenge test. In a study (Fail et al . 1996a)
with consecutive in vivo and ex vivo portions, (and with hCG challenge in the ex vivo portion), in

vivo 175 mg/kg/day iprodione treatment for 14 days yielded generally sim ilar results to other in

vivo rat studies ( e.g. , decreased androgen-sensitive tissue weights, increased adrenal gland and relative testes weights and increase d plasma LH levels) at a dose th at showed overt toxicity (10% decrease in body weight). In the ex vivo portion, iprodione decreased te stosterone secretion from testicular sections in a dose-dependent manner, with or without hCG stimulation; no differences were observed due to the 14-day iprodione in vivo exposure. In another study (Fail et al ., 1996b)
with in vivo hCG challenge, 14-day ip rodione treatment with 175 mg/kg/day produced similar results as described above, incl uding evidence of overt toxicity (13% decrease in body weight).
No differences were observed in testicular or plasma testosterone levels after in vivo hCG challenge.

Finally, two special studies examined effects on Leydig cells after iprodione treatment. An

in vivo rat study designed to examine iprodione treat ment effects on Leydig cell proliferation (Kennel, 2002a) demonstrated a dose-dependent response at ≥ 70 mg/kg/day iprodione, with a possible treatment-rela ted effect as low as 6 mg/kg/day. In an in vitro study using cultured porcine Leydig cells (Benham ed, 1995), exposure to iprodione or two iprodione metabolites
resulted in a reversible inhibiti on of hCG-stimulated testosterone secretion. The results of these studies complement previous fi ndings, and demonstrate that ip rodione-induced effects on Leydig cell (proliferation and inhi bition of testosterone s ecretion) are reversible.IPRODIONE FINAL

Page 17 of 66 As stated previously in the estrogen pathway section, in avian reproduc tion studies, there was a reduction in duck hatchling surviv al at Day 14 and a reduction in quail hatchling body weight at the high dose (1000 ppm). However, the reduc tion in mallard reproduction was observed along with a 5 mallards exhibiting loss of coordination or wing droop.IPRODIONEFINALPage 18 of 66Table 3. Androgenic/Anti-Androgenic Pathway for IprodioneLines of Evidence Indicating Potential Interaction withthe Androgenic/Anti-Androgenic Pathway for Iprodione1Study Type / LiteratureCitation AR Binding / AR activation Steroidogenesis Sex Steroid Hormones Testes Weight/GSI Gonad Staging / Histopathology Epididymides Weight Epididymides Histopathology Pituitary Weight Accessory Sex Organ Weights / 2° Sex Characteristics Fertility(Frt) / Fecundity (Fcd) / Reproductive success (Repro) Age and Weight at PPS Vitellogenin (VTG) Systemic Toxicity Observed 2 Overt Toxicity Observed 3EDSP Tier 1 AssayAR BindingRequirement satisfied by OSRI (multiple studies see below)SteroidogenesisRequirement satisfied by OSRI (multiple studies; see below)HershbergerRequirement satisfied by OSRI (multiple studies; see below)Male Pubertal RatRequirement satisfied by OSRI (multiple studies; see below)FSTRA(MRID 48617204)NENE4NFrt:↓12%(H)NNOSRIARTA(Kojimaet al., 2004)NAR Binding(Blystoneet al., 2007and 2009; Fail et al.,1994)PSteroidogenesis(Blystoneet al., 2007)PIPRODIONEFINALPage 19 of 66Table 3. Androgenic/Anti-Androgenic Pathway for IprodioneLines of Evidence Indicating Potential Interaction withthe Androgenic/Anti-Androgenic Pathway for Iprodione1Study Type / LiteratureCitation AR Binding / AR activation Steroidogenesis Sex Steroid Hormones Testes Weight/GSI Gonad Staging / Histopathology Epididymides Weight Epididymides Histopathology Pituitary Weight Accessory Sex Organ Weights / 2° Sex Characteristics Fertility(Frt) / Fecundity (Fcd) / Reproductive success (Repro) Age and Weight at PPS Vitellogenin (VTG) Systemic Toxicity Observed 2 Overt Toxicity Observed 3Hershberger(Blystoneet al., 2009)NAVP↓30%(H)LABC↓10,11, 21%(L,M,H)↓LW(M,H)↓AWNMale Pubertal Rat(Blystoneet al., 2007)↓73, 83,89%(L, M, H)NNA11%(H)NANASV↓24%(H)↑2 day(M)↑2.8 day(H)↑16% (H)↑LW;↑AW(H)NFSTRA(MRID 48617204)NENE4NFrt:↓12%(H)NNOSRISubchronic oral toxicity(Rat;MRID 42960701)N NNX(M, H)X(M,H)Subchronic oral toxicity(Mouse;MRID 48125502)N N NE N NX(ML,MH, H)X(MH,H)IPRODIONEFINALPage 20 of 66Table 3. Androgenic/Anti-Androgenic Pathway for IprodioneLines of Evidence Indicating Potential Interaction withthe Androgenic/Anti-Androgenic Pathway for Iprodione1Study Type / LiteratureCitation AR Binding / AR activation Steroidogenesis Sex Steroid Hormones Testes Weight/GSI Gonad Staging / Histopathology Epididymides Weight Epididymides Histopathology Pituitary Weight Accessory Sex Organ Weights / 2° Sex Characteristics Fertility(Frt) / Fecundity (Fcd) / Reproductive success (Repro) Age and Weight at PPS Vitellogenin (VTG) Systemic Toxicity Observed 2 Overt Toxicity Observed 3Chronic oral toxicity(Dog, MRIDs00144391,41327001 and42211101)N N NE N NProstate↓22, 41%(M, H)X(H)NChronic oral toxicity /carcinogenicity(Rat;MRID 42787001)↑17–30%(H)(H)NE (H)NProstateatrophy(H)SV:↓(H)X(M, H)NCarcinogenicity(Mouse;MRID 42825002)N(M, H)NE NE NX(H)NSpecial developmentaltoxicity- Rat(Repetto-Larsey, 1997)AGD↓5,14%(M, H)5X(H)Developmental toxicity- Rat(Wolfet al. 1999)N N N N N NNTwo-generationreproduction(Rat;MRID 41871601)NE N NE N NE NENX(M,H)X(M, H)Special Study —hormone synthesis(Chuzel, 1998)↓Test.(L, H);↑LH(H)NIPRODIONEFINALPage 21 of 66Table 3. Androgenic/Anti-Androgenic Pathway for IprodioneLines of Evidence Indicating Potential Interaction withthe Androgenic/Anti-Androgenic Pathway for Iprodione1Study Type / LiteratureCitation AR Binding / AR activation Steroidogenesis Sex Steroid Hormones Testes Weight/GSI Gonad Staging / Histopathology Epididymides Weight Epididymides Histopathology Pituitary Weight Accessory Sex Organ Weights / 2° Sex Characteristics Fertility(Frt) / Fecundity (Fcd) / Reproductive success (Repro) Age and Weight at PPS Vitellogenin (VTG) Systemic Toxicity Observed 2 Overt Toxicity Observed 3Special Study —hormone synthesis(Chuzel, 2001)↓Test.,↑LH(L,H)NSpecial Study —hormone synthesis(Kennels, 2002b)↓Test.,↑LH(L, M, H)NSpecial Study —Testicular effects(Kennel, 2002a)Rel.↑6%(H)↑LeydigcellsNSpecial Study —Testicular effects(Benhamed, 1995)↓Test.Avian reproduction(Duck;MRID 00086840)NNX(H)Avian reproduction(Quail;MRID 00099126)P6(H)N N1. Key to responses: L=Low treatment, ML=Medium-low treatment,M=Medium treatment, MH=Medium-high treatment, H=High treatment.Arrows (↓or↑) indicate the direction of the response. Ashaded cell indicates that is parameter is not routinely evaluated or is not applicable in this assay.Changes in weight are absolute unless otherwise indicated. Changes in weight are absolute unless otherwise indicated. Abbreviations for androgen sensitivetissues: Seminal vesicles (SV), Ventral prostate (VP), Dorsal prostate (DP), Prostate(P), Levator ani-bulbocavernosus (LABC),Epididymides (E).LW = Liver weight; AW = Adrenal weight; Test = testosterone; LH = luteinizing hormone2. The systemic toxicity in the Tier 1 assays are presented in this column (e.g. KW= kidney weight). The systemic toxicity for the OSRI is indicated by an X inthis column. For details see Section IV. AIPRODIONEFINALPage 22 of 663. The overt toxicity in the Tier 1assays are presentedin this column (e.g.↓BW). The overt toxicity for the OSRI is indicated by an X in this column. Fordetails see Section IV. A4. The only histopathological changes noted in gonads of treated males were slight increases in the incidence of minimal to mild interstitial cell hyperplasia andasynchronous gonad development.5. A dose response was observed for the decrease in AGD, however this effect co-occurswith overt toxicity at the high dose (250 mg/kg/day).6. Reduction in duck hatchling survival at Day 14 and a reduction in quail hatchling body weight at the high dose (100 mg/kg-diet)E EquivocalN Negative (in vitro)/No effect (in vivo)NA Not applicableNE Not examinedIPRODIONE FINAL

Page 23 of 66 C. Effects on Hypothalamic-Pitui tary-Thyroidal (HPT) Axis

The current EDSP Tier 1 batte ry does not have a specific in vitro assay to detect chemicals with the potential to affect hypothalamic or pituitary regulation of thyroid hormone production, but it
does include three in vivo assays that provide data to de tect changes in the HPT axis, i.e ., the pubertal female and male (rat) assays, and the AMA (frog).

The potential for iprodione to interact with thyroid regulation is summarized in Table 4. The table depicts the lines of evidence indicating pot ential interaction with the pathway. The various targets of the thyroid pathway across the relevant Tier 1 assays are delineated to facilitate determination of the potential for thyroid or HPT axis effects. This table also includes HPTrelevant findings from data ev aluated as OSRI. This table also includes HPG-relevant findings
from data evaluated as OSRI. Effects that occurred in the presen ce of overt toxicity are discussed in the text but are not reported in the table and not considered fu rther in the WOE assessment.

In the AMA, no effects were noted on the deve lopmental stage, thyroid histopathology, hind limb length (normalized), or on growth endpoints (body weight and snout-vent length). In the female pubertal assay, serum T 4 concentrations were decreased by ≥ 93% (p<0.05) at both doses and pituitary weights (absolute and relative) were decreased (p <0.05) by 12% at the high dose, compared to control; however no changes in thyroid weight or histopathology, or serum TSH concentration were noted. Overt toxicity was no t observed in this study, but increases in liver weights (9-14% at high dose) and adrenal weight s (24-42% at mid and hi gh dose) were observed. In the OSRI used to satisfy the Tier 1 male pubertal assay ( e.g . Blystone et al ., 2007), effects on the thyroid pathway were not evaluated. In th e agency’s evaluation of the OSRI (EDRT report 2009), measures of the potential of iprodione to di srupt the HPT axis in pubertal males will be provided by the female pubertal assay.

Thyroid and pituitary weight and histology we re evaluated in the Part 158 subchronic and chronic rat and mouse studies, a nd the chronic dog study. After treatment with iprodione, thyroid weight was not changed in the subchronic mouse study or the chronic dog study. In the subchronic rat study, thyroid and pituitary weights were decreased (42% and 20–24%, respectively) at the high dose, in the presence of overt toxicity; pituita ry weights were also
decreased 35–48% at the middle dose. In both the chronic rat study and the mouse carcinogenicity study, thyroid weights were in creased by 14 to 18% in males. However, histopathological changes in the thyroid were not present in males or females in any in vivo
studies and no treatment-related ch anges were noted in pituitary weight in the subchronic mouse study, chronic dog or chronic rat studies, or the mouse carci nogenicity study.IPRODIONE FINAL

Page 24 of 66 Table 4. Thyroid Pathway for Iprodione.

Lines of Evidence Indicating Potential Interaction with the Thyroid Pathway for Iprodione 1

Study Type / Literature CitationThyroid WeightThyroid: Gross andHistopathologySerum T4LevelsSerum TSH LevelsPituitary WeightDevelopmentalstage orasynchronous &NHLLGrowth(BW,SVL,)Systemic Toxicityobserved2Overt ToxicityObserved3

EDSP Tier 1 Assays

Male Pubertal Rat EDRT Report: Measures of the potential to disrupt the HPT axis will be provided by the Female Pubertal Assay.

Female Pubertal Rat (MRID 48671502) N N ↓ 97% (L) ↓ 93%

(H) N ↓ 12% (H) N ↑ LW (H); ↑ AW (H) N

AMA (Frog; MRID 48671501) N N N N

OSRI

Subchronic oral toxicity (Rat; MRID 42960701) N N ↓ 15% (females ; L) X (M, H) X (M, H)

Subchronic oral toxicity (Mouse; MRID 48125502) N N N X (L, ML, MH, H) X (ML, MH, H)

Chronic oral toxicity (Dog; MRIDs 00144391, 41327001 and 42211101) N N N X (H) N

Chronic oral toxicity / carcinogenicity (Rat; MRID 42787001) ↑ 14% (male,

H) N N X (M, H) N

Carcinogenicity (Mouse, MRID 42825002) ↑ 18% (male,

H) N N X (H) N

1. Key to responses: L=Low treatment, ML=Med ium-low, M=Medium treatment, MH=Medium-high, High treatment (H). Arrows ( ↓ or ↑ ) indicate the direction of the response. A shaded cell indicates that this para meter is not routinely evaluated or is not applicable in this assay. 2. The systemic toxicity in the Tier 1 assays are presented in this column ( e.g . KW= kidney weight). The systemic toxicity for the OSRI is indicated by an X in this column. For details see Section IV. A 3. The overt toxicity in the Tier 1 assays are presented in this column ( e.g

. ↓ BW). The overt toxicity for the OSRI is indicated by an X in this column. For details see Section IV. A P Positive findings N Negative findingsIPRODIONE FINAL

Page 25 of 66 IV. Committee’s Assessment of Weight of Evidence

This section of the document describes the we ight of evidence (WoE ) determination on the potential of iprodione to interact with endocrine related processes ( i.e., E, A or T hormonal pathways) as well as recommendati ons regarding Tier 2 testing. The results of the Tier 1 assays are considered, along with other scientifically relevant information ( e.g., 40 CFR Part 158 test guidelines and published or publicly available pe er-reviewed studies). WoE analysis in the context of the EDSP follows the Agency’s gui dance (USEPA 2011) and is conducted on a caseby-case basis by first assessing the different lines of evidence ( i.e., specific Tier 1 assays and OSRI), then performing an integrated analysis of those lines of evidence.

The WoE evaluation includes consid erations of biological plausibility of the findings from the different lines of evidence by examining the co nsistency, coherence, and interrelationships among the measured endpoints within and across studies. The available findings from standard toxicology studies on the substance may contribute to the WoE evalua tion in helping elucidate if effects seen in the Tier 1 assay are relate d to perturbations of the endocrine system per se or alternatively a sequelae of system ic effects. Endocrine modes of action may elicit a number of phenotypic consequences other than thos e evaluated in the Tier 1 assays.

Endocrine-related findings in the presence of overt toxicity may result in uncertainty as to whether or not the responses ar e related through an endocrine pathway, therefore non-endocrine toxic responses (including but not limited to mortality or body weight changes) are also
considered in this WoE evaluation. The FIFRA SAP that evaluated scientific issues associated with weight of evidence eval uation of the results of the Tire 1 assays stated that “In summary, the Panel agreed that little, if any, weight shoul d be placed on signs of endocrine disruption in the presence of overt toxicity. A ll effects in endocrine sensitive tissues should be evaluated in terms of primary interactions with the endocrine system vs. secondary effect s related to toxicity in non-endocrine organs or overal l disruptions in homeostasis” (USEPA, 2013).

A. Systemic/Overt toxicity in the in vivo Tier 1 Assay and OSRI

Effects that were considered to be systemic or overt toxicity for the in vivo Tier 1 assays and OSRI studies are described belo w. In addition to the endocrine -related effects described above for the in vivo Tier 1 assays and OSRI, other eff ects observed which may be considered overt/systemic toxicity are also describe below. Generally, one parameter (i.e., death or >10% decrease in mammalian body weight) was sufficient for a dose/concentration to be considered overtly toxic. However, in other instances, mo re than one parameter was needed to determine overt toxicity. Effects which were considered to be signs of system ic toxicity were generally less severe form s of toxicity (e.g., changes in organ weights or blood chemistry).IPRODIONE FINAL

Page 26 of 66 1. Tier 1 Assays

The requirements for Hershberger and the male and female pubertal assays were satisfied by OSRI. The systemic/overt toxicity observed in the OSRI studies are discussed below. In the uterotrophic assay, body weights we re decreased (p<0.05) on Day 4 ( ↓ 12%) in the high dose group compared to the vehicle cont rols, resulting in a decrease (p<0.05) in overall body weight gain ( ↓ 54%).

2. OSRI

Blystone et al ., 2009 (MRID 48279202) reported significant incr eases in liver weight at 100 to 200 mg/kg ( ↑ 6 to 10%; p<0.05) and in adre nal weights at 200 mg/kg ( ↑ 44%, p<0.001).

In a subchronic rat feeding study when admi nistered in the diet at 0, 78, 151, 252, and 355 mg/kg/day for males and 89, 189, 266, and 408 mg /kg/day for females, iprodione caused substantial reductions in body weight ( ≥ 24%) and body weight gains ( ≥ 30%) at the medium and high doses and the high dose group was terminated early (Week 8) due to overt toxicity (body weight loss, decreased food cons umption, and clinical signs). Absolute pituitary weight was decreased (p<0.05) in males by 20 and 35% at th e medium and high doses, respectively, and in females by 15, 24 and 48% at the low, medium an d high doses, respectively, with a significant decrease (p<0.05) in females in the relative weight ( ↓ 26%) at the high dose. In females, absolute uterus weight was decreased (p<0.01) by 74% at the high dose, and absolute ovary weight was decreased (p<0.01) by 21 and 51% in the mid- and high-dose groups. Relati ve liver weights at the medium and high doses were increased (NS) by 15 and 32% in males, and 26 and 29% in females. Histopathological lesions were seen in the testes, epididymides, seminal vesicles, prostate, uterus, ovaries and the adrenal glands.

In a subchronic mouse feeding study, iprodione was administered in the diet with at 0/0, 260/327, 509/659, 1066/1303, or 2130/2608 mg/kg/day for males/females for 13 weeks. Adjusted (for body weight) liver weights were do se-dependently increased (p<0.01) in males at the mid-low ( ↑ 26%), mid-high ( ↑ 98%) and high dose ( ↑ 133%) and in females at the low ( ↑ 19%) mid-low ( ↑ 41%), mid-high ( ↑ 98%) and high dose ( ↑ 149%). Absolute adrenal weights were also dose-dependently increased (p<0.01) in males at the mid-high ( ↑ 28%) and high dose ( ↑ 32%), and in females at the low ( ↑ 28%) mid-low ( ↑ 31%), m id-high ( ↑ 48%) and high dose ( ↑ 30%). Also at the high-dose group, adjusted kidney weights were decreased (males, ↓ 31%; females, ↓ 17%) and absolute spleen wei ghts were increased (males, ↑ 15%; females ↑ 45%). Decreases
were also noted in absolute uterus wei ght in the mid-high- and high-dose groups ( ↓ 29, p<0.05 and 58%, p<0.01) and in absolute and adjusted ovary weights ( ↓ 50–53%, p<0.01) in the high-dose group. No treatment-related changes were se en in absolute or adjusted weights of the testes, pituitary or thyroid glands at any dose. Grossly, the liver was en larged with irregularIPRODIONE FINAL

Page 27 of 66 surface depressions in most animals in the mid-hi gh- and high-dose groups. Histological lesions were seen in the liver, uter us, ovaries and adrenal glands.

In a chronic toxicity study with dogs, ip rodione at the high-dose (male 145.3/female 152.5 mg/kg/day), caused increases in absolute and relative liver weight s [males, 41% and 48%; females, 18% (NS) and 28%] and in alkaline phosphatase, SGOT, SGPT and LDH enzymes in both sexes. In addition, absolute and relative adrenal weights we re increased (p<0.05 or p<0.01) in high-dose males (37% and 42%) and females ( 43% and 51%). Males also had decreases in both absolute (mid-dose, ↓ 22%, p<0.05; high-dose, ↓ 41%, p<0.01) and relative (high-dose, ↓ 39%, p<0.01) prostate weights.

In the combined chronic toxicity/ carcinogenicity study in rats, when administered in the diet at 0, 6.1, 12.4, and 69 mg/kg/day to males and 0, 8.4, 16.5, and 95 mg/kg/day to females body-weight gains were decreased in both sexes at the high-do se compared to control. Overall, body-weight gains decreased by 14% and 8% in the high-dose males and fe males, respectively; and body weight gains at Week 12 were decreased by 14% in high-dose males and by 19% in high-dose females. Food consumption was also decreased sli ghtly at this dose in both sexes. Treatmentrelated changes were seen in absolute liver we ights but adjusted liver weight was increased by 9% (p<0.05) and 11% (p<0.01) in mid- and hi gh-dose males, respectively. At Week 52, highdose males displayed an increase in the incidenc e of lesions in the adrenals, and there was anincreased incidence of centrilobular hepatocyte enlargement in mid- and high-dose males. Highdose females displayed an increase in centrilobula r hepatocyte enlargement and an increase in the incidence of generalized rare faction and fine vacuolation of the zone fasciculata in the adrenals compared to the control and other dose groups.

In a carcinogenicity study, iprodione was administ ered in the diet to mice at doses of 0, 23, 115 and 604 mg/kg/day for males and 0, 27, 138 and 793 mg/kg/day for females. Absolute and relative adrenal weights were increased at the high dose at Week 52 in males ( ↑ 67–78%, p<0.01) and females ( ↑ 14–17%, NS) and at Week 100 in males ( ↑ 14–21%, NS) and females ( ↑ 6–14%, NS). At terminal sacrifice, adrenals we re enlarged in high-dose males (4/50, 8%) and females (5/50, 10%) compared to control males (1/50, 2%) and females (0%). In addition, the majority of high-dose females (12/13) displayed hype rtrophy of the cells of the zona fasciculate; this lesion was not seen in any other dose group. Liver weights (abs olute, adjusted, and relative) were generally increased in a dose-dependent manner, but the increases only attained significance (p<0.01) at 52 weeks fo r adjusted liver weights (male ↑ 52% and female ↑ 66%) and at 100 weeks for absolute liver weight in females ( ↑ 2.81-fold) and adjusted liver weights in males ( ↑ 2.88-fold). There was an increase (p<0.01) in the incidence of liver tumors in both males (26/50) and females (21/50) at the high-dos e compared to control (male, 7/50; females, 2/50), which was accompanied by increases in seve ral liver lesions (cen trilobular hepatocyte enlargement/vacuolation, area(s) of enlarged eosinophilic hepatocytes, pigm ented macrophages,IPRODIONE FINAL

Page 28 of 66 centrilobular necrosis, and amyloid deposits). SGOT and SGPT concentrations were elevated at the high-dose in both sexes compared to cont rol at the interim sacrifice (only time these enzymes were examined).

In a special developmental toxicity study (Repetto-Larsey, 1997) following gavage administration to rats at doses of 0, 20, 120, or 250 mg/kg/day during GD 6 through 19, deaths occurred (9/25) in addition to decreased body-weight gain an d food consumption/efficiency.

In the avian reproduction studies, th e mallard exhibited significant ove rt toxicity in the form of loss of coordination and wing droop.

Overt toxicity was observed in a fathead minnow early life stage study at the mid-high and high exposures (1.1 and 2.2 mg/L), based on decreases in larval survival ( ↓ 42 and 96%). Additionally, wet body wei ght was decreased ( ↓ 26 and 70%) as well as body length ( ↓ 11 and 31%).

B. Estrogen Pathway

For the estrogen pathway, the Tier 1 in vitro aromatase, ER binding, ERTA and the in vivo uterotrophic assays were negative. However, there appears to be a potential for in teraction with the estrogen pathway characterized by effects seen in the female pubertal assay (delay in VO, delay in age at first estrus a nd uterine histopathology) as well as OSRI studies (uterus atrophy and decreased number of corpora lutea). Decreases in fecundi ty/fertility or reproductive/offspring parameters were observed in fish (FSTRA) and birds (Part 158 bobwhite quail study), however, it is noted that these e ffects have limited diagnostic utility for understanding potential interacti on with the estrogen pathway. Ba sed on the evidence from male assays (OSRI), this potential interaction with th e estrogen pathway may be due to alteration of the steroidogenesis pathway.

C. Androgen Pathway

The overall results of the OSRI ( in vitro AR binding/ARTA and steroidogenesis, and in vivo Hershberger and pubertal male rat study) and the FSTRA, indicate a potential for interaction with the androgen pathway. These effects include d decreases in the androgen-sensitive organ weights in the Hershberger a ssay, delays in age and weight at PPS and decreased serum testosterone and androgen-sensitiv e organ weights in the male pubertal assay, as well as similar results in the Part 158 mammalian toxicity and special mechanistic studies. Effects on male fish gonadal hi stopathology and effects on fertil ity were observed as well as reproductive effects in birds; again, it is noted th at effects on fecundity an d fertility have limited ability for informing directi on interaction with the androge n pathway. Although iprodione did not demonstrate competitive binding to the AR, ip rodione metabolites (produced in cell culture)IPRODIONE FINAL

Page 29 of 66 showed evidence of direct interaction with the AR. Additionally, there is evidence of iprodione potentially impacting steriodogenesis. Mechanis tic studies and studies published in the open literature suggests that iprodione may interfere with the active transport of cholesterol, the biosynthetic precursor of testoste rone, into Leydig cells, thereby re sulting in potential decrease in overall steroid hormone levels, in cluding testosterone. Evidence for steroidogenesis inhibition is demonstrated by alterations in hormone levels (testosterone, LH progest erone and aldosterone), decreases in androgen-sensitive tissues weight s, delayed PPS and increased adrenal gland weights in mammals.

D. Thyroid Pathway

In the female pubertal assay, serum T 4 concentrations were redu ced at 150 and 300 mg/kg/day ( ↓ 97% and ↓ 93%); however, there were no effects on TS H, thyroid weight or histopathology. Additionally, no thyroid related effects were observed in the Part 158 mammalian toxicity studies. No thyroid-related effects were noted in the AMA. Therefore, there is no convincing evidence of an interaction with the thyroid pathway.

E. Conclusions

Based on the weight of the evidence for iprodion e, there appears to be a potential to alter steriodogenesis which may affect the estrogen and androgen pathways in mammals and wildlife. Iprodione does not appear to potentially interact with the thyroid pathway.

V. EDSP Tier 2 Testing Recommendation

Overall, based on the weight of the evidence there appears to be a potential for iprodione to alter steriodogenesis, which may affect the estrogen and androgen pathways in mammals and wildlife.

For iprodione a point of departure (POD) of 0.5 mg/kg/day is used for human health risk assessment. The study by Blystone et al. (2007) was accepted to satisfy the requirement for the male pubertal assay. In that study there was a dose-dependent decr ease in testosterone levels in males at 50 mg/kg/day, the lowest obs erved effect level (LOAEL). Therefore, a 10X Uncertainty Factor (UF L ) was applied ( i.e ., for the use of a LOAEL) resulting in an extrapolated NOAEL of 0.5 mg/kg/day ( i.e , the POD). This POD is used for establishing the chronic Reference Dose (RfD) and to calculate the Margins of Exposure (MOEs) for dietary and non-dietary exposure risk assessments, re spectively. The POD is approximately 300-fold lower than the lowest dose (150 mg/kg/day) that caused estrogen-related effects in the female pubertal assay. Consequently, the ch ronic RfD and the MOEs are likely to be protective of the
endocrine-related effects s een in the pubertal assay.IPRODIONE FINAL

Page 30 of 66 Therefore, mammalian EDSP Tier 2 testing is no t recommended for iprodione since additional testing is not expected to impact EPA’s current regulatory point of departures and endpoints for human health risk assessments.

For wildlife, the current chronic fish NOAEC value is 0.26 mg/L based on a fathead minnow early life-cycle study (LOAEC is 0.55 mg/L base d on survival). Eff ects on fecundity were
observed in the FSTRA at ≥ 0.084 mg/L. Furthermore, effects on fertility were observed at 0.60 mg/L and potential treatment-re lated effects on male gonadal hi stopathology were observed at 0.084 mg/L. Therefore, the EDSP Tier 2 Meda ka Extended One Generation Reproduction Test (MEGORT) is recommended.

For birds, Part 158 avian reproduction studies w ith both northern bobwhite quail and mallard duck are available for iprodione. In both spec ies, effects on reproductive parameters were observed. The type of data obtained from Pa rt 158 avian reproduction studies (OCSPP 850.2300) are considered sufficient for evaluating potential reproductive effects to birds from iprodione exposure. Relevant to EDSP, additional testing is not recommended.IPRODIONE FINAL

Page 31 of 66 VI. References

Benhamed, M. (1995). Effects of iprodione and its metabolites on testosterone secretion in cultured Leydig cells. INSERM U 407, Sophi a Antipolis Cedex, France. INSERMIU 407/95001, R014672. Unpublished study. (MRID 43830601).

Benhamed, M. (1996). Effects of iprodione and its metabolites RP 36112 and RP 36115 on testosterone secretion in cultu red Leydig cells: Sites of action. INSERM U 407, Sophia Antipolis Cedex, France. INSERMIU 407/96001, C024294. Unpublished study. (MRID 44171901).

Blacker, A. and Broadmeadow, A. (1989). Ip rodione: 52 Week Toxici ty Study in Dietary Administration to Beagle Dogs-Addendu m to Study Asssigned MRID No. 00144391: Lab Project Number: NS 12/14. Unpub lished study prepared by Life Science Research, Ltd. in association with Rhone-Poulenc Ag Co. 51 p. (MRID 41327001).

Blystone, C.R., Lambright, C.S., Furr, J., Wils on, V.S., and Gray Jr., L.E. (2007). Iprodione delays male rat pubertal development, reduces serum testosterone levels, and decreases ex vivo testicular testosterone production. Toxicol. Lett. 174: 74–81. (MRID 48279201).

Blystone, C.R., Lambright, C.S., Cardon, M.C., Furr, J., Rider, C.V., Hartig, P.C., Wilson, V.S., and Gray, Jr., L.E. (2009). Cumulative and antag onistic effects of a mixture of the antiandrogens vinclozolin and iprodione in the pubertal male rat. Toxi col. Sci. 111: 179–188. (MRID 48279202).

Breier, J.M., Olmstead, A., Bowers , L.M. (2012) weight of evidence to evaluate the potential for iprodione to affect estrogen, androgen or thyroid systems. Ba yer CropScience, Arlington, V. Report No. USO277, MRID# 48685301

Broadmeadow, A. (1984). Iprodione: 52-week toxici ty study in dietary administration to Beagle dogs. Life Science Research Ltd., Eye,Suffol k, England. C021579. Unpublished study. (MRID 00144391).

Chambers, P.R, Crook, D., Gibson, W.A., Gopina th, C, and Ames, SA (1992). Iprodione: Potential tumorigenic and toxic effects in prolon ged dietary administrati on to rats. RhonePoulenc Agrochimie, Lyon, France. R014586. Unpublished study. (MRID 42637801).

Chambers, P.R, Crook, D., Gibson, W.A., Read, RM., and Gopinath, C. (1993). Iprodione: Potential tumorigenic effects in prolonged di etary administration to mice. RhonePoulenc Agrochimie, Lyon, France. C021564. Unpublished study. (MRID 42825002).

Chuzel, F. (1998). Hormonal Measurements in Adult Male Sprague-Dawley Rats Following Administration of Iprodione by Gavage. Rhone-P oulenc Agro, Centre de Recherche. Study No. SA 98283. Unpublished study. (MRID 44729201).IPRODIONE FINAL

Page 32 of 66 Chuzel, F. (2001). Iprodione: Hormonal measur ements in adult male Sprague-Dawley rats following a single administration of iprodione by gavage. Av entis Crop Science, Sophia Antipolis Cedex, France. Study No. SAOI OI6, C019327. Unpublished study. (MRID 45859601).

Davis, J.P. (2012). The Uterotrophic A ssay (OPPTS 890.1600) for Iprodione. Unpublished study conducted by Integrated Laboratory Systems, Inc., Durham, NC. La boratory Project Study No.: C199-100, January 3, 2012. (MRID 48617205).

Davis, J. P. (2012). Pubertal Development and Thyroid Function in Intact Juvenile/Peripubertal Female Rats for Iprodione. Unpublished conduc ted by Integrated Laboratory Systems, Inc., Durham, NC. Laboratory Report N o. C199-300, April 3, 2012. (MRID 48671502).

Fail, P.A., Anderson, S.A., and Pearce, S.W. (1994). Toxicity testing of a fungicide, iprodione in adult male CD Sprague Dawley rats. Laborator y of Reproductive Endocrinology, Center for Life Sciences and Toxicology, Research Triangle Institute, Research Triangle Park, NC. RTI Identification No. 65C-5703, C024395. Unpublished study. (MRID 43535002).

Fail, P.A., Anderson, S.A., and Pearce, S.W. (1996a). Toxicity testing of a fungicide, iprodione: Endocrine toxicology studies of testes from adult male CD Sprague-Dawley rats exposed to iprodione in vitro. Laboratory of Reproductive and Endocrine Toxicology, Research Triangle Institute, Research Triangle Park, NC. RTI Identification No. 65C-6169; Rt95-IPDA; RTI-532, C024292. (MRID 44171903).

Fail, P.A., Anderson, S.A., and Pearce, S.W. (1996b). Toxicity testing of a fungicide, iprodione: Endocrine toxicology studies of testes from adult male CD Sprague-Dawley rats exposed to iprodione in vivo. Laboratory of Reproductive and Endocrine Toxicology, Research Triangle Institute, Research Triangle Park, NC. RTI Identification No. 65C-6169; Rt95-IPDA; RTI-527, C024293. Unpublished study. (MRID 44171904).

Fink, R.; Beavers, J.B.; Joiner, G.; et al. ( 1981). Final Report One- generation Reproduction Study—Mallard Duck: Iprodione Technical: Project No. 171-103. Unpublished study received Oct 26, 1981 under 46153-1; prepared by Wildlife International, Ltd., and Rhone-Poulenc, Inc., submitted by Precision Compounding, Inc., Newa rk, N.J.; CDL:246150-C. (MRID 00086840).

Fink, R.; Beavers, J.B.; Joiner, G.; et al. ( 1981). Final Report: One- generation Reproduction Study —Bobwhite Quail: Iprodione Techni cal: Project No. 171-102. Unpublished study received Oct 26, 1981 under 46153-1; prepared by Wildlife Internati onal, Ltd. and RhonePoulenc, Inc., submitted by Precision Com pounding, Inc., Newark, N.J.; CDL:246150-B. (MRID 00099126).

Fryer, S.; Duckworth, J.; Gibson, W.; et al. (1993 ). Iprodione Sub-Acut e Toxicity to Rats by Dietary Administration for 13 Weeks: Amended Report: Lab Project Number: RNP 322/90767. Unpublished study prepared by Huntingdon Re search Centre Ltd. 214 p. (MRID 42960701).IPRODIONE FINAL

Page 33 of 66 Genwood, S.M. (1991). Two-genera tion reproduction study with ip rodione technical in rats. Hazleton Laboratories America, Inc., Madis on, WI. HLA Identifica tion No. HLA 6224-154. Unpublished study. (MRID 00162983).

Kangas, L. (1991). A 52-week dietary toxi city study of iprodione in the Beagle dog. BioResearch laboratories Ltd., Senneville, Queb ec, Canada. Laboratory project identification No. 84296, C021573. Unpublished study. (MRID 4221110).

Kennel, P. (2002a). Measurement of Leydig cell proliferation following a 14-day treatment period with iprodione by gavage in the rat. Ba yer CropScience, Sophia Antipolis Cedex, France. Study No. SA01315, C025273. Unpublished study. (MRID 45859602).

Kennel, P. (2002b). Iprodione: Hormonal measurements following a 14-day treatment period with iprodione by gavage in the rat. Bayer CropScience, Sophia Antipolis Cedex, France. Study No. SA01287, C024675, July 12, 2002. Unpublished study. (MRID 45859603).

Kojima, H., Katsura, E., Takeuchi, S., Niiyami, K., and Kobayashi, K. (2004) Screening for estrogen and androgen receptor activi ties in 200 pesticides by in vitr o reporter gene assays using Chinese hamster ovary cells. Environ. Health Perspect . 112 (5): 524-531.

Lee, M.R. (2012). Iprodione-Amphibian Meta morphosis Assay with African Clawed Frog ( Xenopus laevis ) Following OPPTS Test Guideline 890.1100 and OECD Test Guideline No. 231. Unpublished study performed by Smithers Vi scient, Wareham, MA. Smithers Viscient Study No. 13798.6271. Protocol/Project No. EBIDP002. Study sponsored by Bayer CropScience, Research Triangle Park, NC. March 28, 2012. (MRID 48671501).

Repetto-Larsay, M. (1997). Iprodione: Toxicology study in pregnant rat by gavage to examine sex differentiation. Rhone-Poulenc Agro CRS A, Cedex, France. Study No. SA96448, C022658. Unpublished study. (MRID 44365001).

Rodwell, D. (1985). A Teratology Study in Rabb its with Iprodine: Fi nal Report: Project No. WIL-21028. Unpublished study prepared by WIL Research Laboratories, Inc. 158 p. Unpublished study. (MRID 00155469).

Surprenant, D. (1988). The Toxicity of Iprodione Technical to Fathead Minnow ( Pimephales promelas ) Embryos and Larvae: Rept. No. 88-2-2639. Unpublished study prepared by Springborn Life Sciences, Inc. 59 p. (MRID 40550801).

Tesh, J.M., McAnulty, P.A., Deans, C.F., Wilby, O.K., and Tesh,SA (1986). Iprodione (technical grade): Teratology study in the rat. Rhone-Poulenc Agrochimie, Lyon, France. C023030. Unpublished study. (MRID 00162984).

Tesh, J.; McAnulty, P.; Lambert, E.; et al. (1987). Iprodione (Techni cal Grade): Teratology Study in the Rat: Supplementary Litter Data: Laboratory Identifica tion No. 87/RHA064/755. Unpublished study prepared by Life Sc ience Research. 95 p. (MRID 40514901).IPRODIONE FINAL

Page 34 of 66 USEPA (2010). Iprodione — Report of the Endoc rine Disruptor Review Team·Test Order #: EDSP-I09801-174. December 16, 2010. Office of Chem ical Safety and Pollution Prevention. U.S. Environmental Protection Agency, Washington, DC.

USEPA (2011). Endocrine Disruptor Screeni ng Program — Weight of Evidence: Evaluating Results of EDSP Tier 1 Screening to Identify th e Need for Tier 2 Testing. Office of Chemical Safety and Pollution Prevention Science Coordina tion and Policy. U.S. Environmental Protection Agency, Washington, DC.

USEPA (2013) Tran s mittal of Meeting Minutes of the FIFRA Scientific Advisory Panel Meeting held July 30 - August 1, 2013 on Weight-of

- Evidence: Evaluati ng Results of EDS P Tier 1 Screening. Office of Chemical Safety and Pollution Pr evention. U.S. Environmental Protection Agency, Washington, DC.

Wilga P.C. (2011). Iprodione: Human Recombinant Aromatase Assay. Unpublished study conducted by CeeTox, Inc., Kalamazoo, MI. Laboratory Report No.: 9077V-100375AROM, December 20, 2011. (MRID 48617201).

Willoughby, J.A. (2011). Iprodione: Estrogen Re ceptor Binding (Rat Uterine Cytosol). Unpublished study conducted CeeTox, Inc., Ka lamazoo, MI. Laboratory Study No.: 9077V100375ERB, December 21, 2011. (MRID 48617202).

Willoughby, J.A. (2011). Iprodione: Estrogen Recep tor Transcriptional Activation (Human Cell Line (HeLa-9903)). CeeTox, Inc., Kalam azoo, MI. Laboratory Report No.: 9077V100375ERTA, December 21, 2011. Unpublished. (MRID 48617203).

York, D. (2012). Iprodione — Short-Term Re production Assay with the Fathead Minnow ( Pimephales promelas ) Following OPPTS 890.1350 and OECD 229 Guidelines. Unpublished study performed by Smithers Viscient, Ware ham, MA. Lab Study No.: 13798.6272, Bayer Study No. EBIDP001. Study sponsored by Bayer CropScience, Research Triangle Park, NC. January 5, 2012. (MRID 48617204).IPRODIONE FINAL

Page 35 of 66 APPENDIX 1. EDSP Tier 1 Screening Assays

Amphibian Metamorphosis Assay (OCSPP 890.1100)

The 21-day assay (MRID 48671501) of iprodione (98.1% purity) on amphibian metamorphosis of African clawed frog ( Xenopus laevis ) was conducted under fl ow-through conditions. Amphibian larvae at Nieuwkoop-Faber (NF) stage 51 (80 per control and treatment group) were exposed to a control and nominal iprodione c oncentrations of 0 (neg ative control), 0.0070, 0.070, and 0.70 mg a.i./L. Mean-measured concentrations were <LOQ (<0.0027, control) 0.0054, 0.066, and 0.61 mg a.i./L. The test system was maintained at 21 to 23°C and a pH of 7.3 to 8.2.

Unless otherwise specified, all conclusions are based on comparison to the negative (FETAX) control. One mortality was observed in the negati ve control, and no mort alities occurred in the treatment groups. Spinal curvature was observed in 41% of control tadpo les and between 14 and 29% of treatment tadpoles with incidence decreasing with increasing mean-measured
concentrations.

Iprodione had no significant effects on developmental [ i.e ., NF stage, normalized hind limb length (HLL)] or growth [ i.e. , HLL, snout-vent length (SVL), wet body weight] parameters on either Day 7 or 21 at any treatment level. No asynchronous development was observed at Day 7 or Day 21. Mild thyroid gland hypertrophy, fol licular cell hypertr ophy, and follicular cell hyperplasia were observed in the co ntrol and all treatment groups; th ese effects did not appear to be treatment-related. Late stage (NF stage > 60) tadpoles were excluded from analyses for continuous endpoints (SVL, we ight, and normalized HLL).

Androgen Receptor (AR) Bi nding Assay (OCSPP 890.1150)

The requirement for a Tier 1 AR bindi ng assay was satisfied by OSRI (Blystone et al ., 2007 and 2009; and Fail et al ., 1994). The results from these studies are presented with the OSRI in Appendix 2.

Aromatase Assay (OCSPP 890.1200)

In an in vitro aromatase (CYP 19) assay (MRID 48617201) , iprodione (98.1% a.i. Lot # AE F062470-01-08/SIN0000217) was incubated with hum an recombinant aromatase and tritiated androstenedione (1- β [ 3 H(N)]-androst-4-ene-3,17-dione; [ 3 H]-ASDN) in ≤ 1% dimethyl sulfoxideIPRODIONE FINAL

Page 36 of 66 (DMSO) at log concentrations of 10

− 11 to 10

− 4 M for 15 minutes to assess the potential of iprodione to inhibi t aromatase activity.

Aromatase activity was determined by measuring the amount of tritiated water produced at the
end of a 15-minute incubation for each concentration of chemical. Tritia ted water was quantified using liquid scintillation counting (LSC). Th ree runs were conducted and each run included a full activity control, a bac kground activity control, a pos itive control series (10

− 10 to 10

− 5 M) using a known inhibitor (4-hydroxyandroste nedione; 4-OH ASDN) in duplicate, and the iprodione series (10

− 11 to 10

− 4 M) with three repetiti ons per concentration.

Aromatase activity in the full activ ity controls averaged 0.616 ± 0.028 nmol

· mg-protein

− 1

· min

− 1

. The response of each full activity control within a run was between 90 to 110% of the average full activity. Activity in the background contro ls ranged 0.24% to 0.39% and averaged 0.30% of the full activity controls.

For the positive control substance (4-OH ASDN), aromatase activity results were within the recommended ranges for the performance criteria. The estimated log IC 50 for 4-OH ASDN averaged

− 7.27 M and the Hill slope was

− 0.94.

Precipitation was noted at the highest c oncentrations of iprodione tested (10

− 3 M); therefore, the maximum concentration used for data analysis was 10

− 4 M. For iprodione, aromatase activity averaged 97.26% and 82.39% of contro l activity at concentrations of 10

− 10 and 10

− 4 M, respectively.

Because the mean aromatase activity for ip rodione was 82.39 ± 0.76% at the highest test
concentration of 10

− 4 M, iprodione is classified as a Non-i nhibitor of aromatase activity in this assay. However, as there is no indication in the st udy report that half-log concentrations such as

10 –3.5 M were evaluated for solubility, there is less confidence in the non-i nhibitor classification. Based on the shape of the res ponse curve, if a soluble 10 –3.5 M concentration had been tested, it is possible that the lowest portion of the curv e would fall within the 50%- 75% activity range resulting in a equivocal clas sification for this chemical.

Estrogen Receptor (ER) Bi nding Assay (OCSPP 890.1250)

In an estrogen receptor (ER) binding assa y (MRID 48617202), uterine cytosol from SpragueDawley rats was used as the source of ER to conduct saturation and competitive binding experiments. A competitive bi nding experiment was conducted to measure the binding of a single concentration of [ 3 H]-17 β -estradiol (1 nM) in the presen ce of increasing concentrations (10

− 11 to 10

− 4 M) of iprodione (98.1% a.i., Lot # AE F062470-01-08 / SIN0000217). DMSO was
used as the solvent vehicle at a final concentrat ion of approximately 4%. A total of three runsIPRODIONE FINAL

Page 37 of 66 were performed, and each run included 19- norethindrone as a weak positive control, octyltriethoxysilane as a negative control, and 17 β -estradiol as the na tural ligand reference
material.

The saturation binding experiment was conducted to demonstrate that the ER in the rat uterine cytosol was present in reasonable numbers and wa s functioning with appropriate affinity for the
radiolabeled reference estr ogen prior to routinely conduc ting ER competitive binding experiments. For this experiment, the appropr iate protein concentration for the saturation binding assay was determined to be in the range of 160-320 μ g/tube, as it bound 25-35% of the [ 3 H]-17 β -estradiol. The mean K d for [ 3 H]-17 β -estradiol was 0.331 ± 0.061 nM and the mean

B max was 0.549 ± 0.100 nM (74.55 ± 3.03 fmol/100 μ g) protein for the prepared rat uterine cytosol. The data produced consis tent and linear Scatchard plots (r 2 >0.98).

In the competitive binding experiment, the estimated mean log IC 50 was

− 9.00 M for the natural ligand (17 β -estradiol) and

− 5.47 M for the weak positive control (19-norethindrone). Compared to the natural ligand, the mean RBA was 0.029% for 19-norethi ndrone. All performance criteria
were met for 17 β -estradiol, 19-norethindrone, and the nega tive control, octyltriethoxysilane.

No precipitation was detected at iprodione concentrations of ≤ 10 –4 M (highest soluble concentration) in any run. Ipr odione was tested over a concentra tion range that fully defined the top of the curve. The mean specific binding of the radiolabeled ligand in the presence of iprodione was >87% at concentrations ≤ 10

− 4 M in all three runs; therefore, an estimated mean log IC 50 and relative binding affinity (RBA) we re not calculated for iprodione.

Based on the results of the three runs, iprodione is classified as Not Interactive in the Estrogen Receptor Binding Assay.

ER α Transcriptional Activatio n (ERTA) Assay (OCSPP 890.1300)

In an estrogen receptor transcript ional activation assay (MRID 48617203), hER α -HeLa-9903
cells cultured in vitro were exposed to iprodione (98.1% a.i. Batch # AE F062470-01- 08/SIN0000217) in logarithmically in creasing concentrations from 10

− 11 to 10

− 4 M in 0.1% DMSO in two independent runs. The experiment s were performed using 96-well plates and each iprodione concentration was tested in triplicate (3 wells/p late) in each run. Cells were exposed to the test agent for 24±2 hours to induce reporter (luciferase) gene products. Luciferase expression in response to activation of the estrogen recepto r was measured using a proprietary luciferase

assay

Iprodione was tested up to and including the limit of cytotoxicity, which was observed at 10

− 4 M in the first run. Cytotoxicity was observed in cells treated with two of the reference compoundsIPRODIONE FINAL

Page 38 of 66 (corticosterone and 17 α -methyltestosterone). A ≥ 20% reduction in cell viability was observed
with 10

− 4 M iprodione, 10

− 5 M 17 α -methyltestosterone and 10

− 4 M corticosterone in the first valid independent run.

In the main assays, the responsiveness of th e cells to the very weak positive control 17 α

methyltestosterone was lower than the expected values, indicating a decrea sed sensitivity of the assay to very weak agonists. Although the conditi ons of this assay were not optimal to detect very weak activity, iprodione responses were similar to those of the negative control corticosterone and not comparable to the responses of 17 α -methyltestosterone, which was able to reach 8.1-52.8% PC. Iprodione was only able to reach a maximum of 0.9- 3.8% PC when tested up to the highest concentration possible ba sed on cytotoxicity. Because the RPC Max < PC 10 in two assay runs, iprodione was considered negative for estrogen receptor transcriptional activation in this test system.

Fish Short-term Reproductive Assay (FSTRA) (OCSPP 890.1350)

The 23-day short-term reproduction assay (MRI D 48617204) of iprodione with fathead minnows ( Pimephales promelas) was conducted under flow-through conditions. Adult fish in 16 spawning groups (2 males and 4 females in each group; 6 m onths old; 1 spawning gr oup per replicate and 4 replicates per treatment group) were exposed to iprodione (technical gr ade, 98.1% purity) at nominal concentrations of 0 (control), 0.0085, 0.085, and 0.85 mg a.i./L; time-weighted average (TWA) concentrations were <LOQ (<0.0030), 0.0086, 0.084, and 0.60 mg a.i./L. The test system
was maintained at 24 to 26 o C and a pH of 6.7 to 7.5. On Day 2, the test solution delivery in replicate C of the 0.60 mg ai/L treatment was rest ricted, resulting in low dissolved oxygen levels and mortality of fish in this replicate; therefor e, this one replicate was excluded from statistical analysis.

There was no significant effect on fish mortality (p>0.05). The ove rall survival in the highest treatment group, 0.60 mg a.i./L, was 72% due to the delivery malfunction which resulted in reduced dissolved oxygen and two male and one fe male mortalities. There were no significant effects (p>0.05) on growth parameters of weight or length in males or females at any treatment level. Secondary sex characteristics (includi ng body color, behavior, body shape, and the presence of the dorsal fat pad) were evaluated, but notable effects were not observed. There were
no treatment-related clini cal signs of toxicity.

Spawning occurred in the control gr oup at least every four days in two of the four replicates and fecundity was 20 eggs/female/day/replicate (ra nge 12.5 to 27.6); fertilization success in the control was 92% (range 89 to 94%). Iprodione significantly (Jonckheere-Terpstra; p<0.05) reduced fecundity 45% and 52% relative to the control at the 0.084 and 0.60 mg a.i./LIPRODIONE FINAL

Page 39 of 66 concentrations. Fertility was al so significantly (Dunnett’s; p<0.05) reduced 12% from control at the 0.60 mg a.i./L treatment level.

There were no significant effects (p>0.05) in the gonadal somatic i ndex (GSI) of male or female fish at any treatment level. Male median nuptial tubercle scores in the iprodione-treated groups did not significantly differ from the control (p>0.05); there were no tubercles noted for females. Plasma vitellogenin (VTG) was not significantly different (p>0.05) in male or female fish at any treatment level compared to the control. Plasma sex steroids were not m easured in this study.

There did not appear to be any effect on gonadal st age development in male or female fish at any treatment level. However, iprodione may have caused some histopathological changes in males, including an increase in the incidence of mi nimal to mild interstitial cell hyperplasia and asynchronous gonad development. Minimal cell h yperplasia was observed in 29% and 20% of male fish in the 0.0086 and 0.60 mg ai/L levels, re spectively, and 43% exhibi ted mild interstitial cell hyperplasia in the 0.084 mg ai/L level, compared to none exhibiting this effect in the control. Similarly, 25% of males in the control exhibited only a minimal condition of asynchronous gonad development, while minimal to mild conditions were noted for 40-43% of male fish in the iprodione-treated groups. No treat ment-related histologi cal changes in females were reported.

Hershberger Assay (OCSPP 890.1400)

The requirement for a Tier 1 Hershberge r assay was satisfied by OSRI (Blystone et al ., 2009). The results from this study are pres ented with the OSRI in Appendix 2.

Female Pubertal Assay (OCSPP 890.1450)

In a Female Pubertal Assay (MRID 48671502), 16 Sprague-Dawley Crl:CD ® (SD) IGS rats/dose group were treated daily via oral gavage with iprodione (98.1%, Batch # AE F062470-01-08) in corn oil at doses of 0, 150 or 300 mg/kg/day fr om post-natal day (PND) 22-42 or 43. Animals were examined for vaginal opening (VO) daily beginning on PND 22 and age and weight at day
of attainment were recorded. After sacr ifice on PND 42 or 43, total serum thyroxine (T 4 ) and thyroid stimulating hormone (TSH) levels were analyzed using a radioimmunoassay. Adrenal, liver, thyroid, pituitary, and urogenital orga n weights were recorded and microscopic examination were performed of the ovaries, uterus, t hyroid, and kidneys.

There were no treatment-related effects on morta lity, clinical signs, or gross pathology. Final body weights and body weight gains were similar to controls across both treated groups.

At 300 mg/kg/day, age at initiation of VO was increased (p<0.05) by 3.5 days (PND 36.3 treated vs. PND 32.8 controls), and age at attainment of VO was incr eased (p<0.05) by 3.9 days (PNDIPRODIONE FINAL

Page 40 of 66 36.8 treated vs. PND 32.9 controls ). Although body weight at atta inment of VO was increased (p<0.05) by 11% at 300 mg/kg/day, th is increase resulted from a co mparison of the weights over an approximately 4-day span (PND 37 treated vs . PND 33 controls). When the 300 mg/kg/day body weights were directly compared to the cont rols on these specific days, mean body weights
were 92.7% of control on PND 33 and 93.4% of c ontrol on PND 37. Therefor e, the delay in age
at initiation and attainment of VO was attributable at least in part to a de crease in body weight at 300 mg/kg/day. Also at this dose, a delay (p<0.05) of 3.5 days (PND 37.2 treated vs. PND 33.7 controls) in the onset of first estr us relative to control was observe d; this delay was attributed to the delay in VO. Mean cycle le ngth was not affected by treatment at either dose level. There was no change in the number of animals cycling; however, there was a significant effect on numbers of animals regularly cycl ing between the treated groups and control (p<0.05). It was stated that the significance of this effect wa s driven by the low number of animals regularly cycling at 150 mg/kg/day (11/ 16) and not by those in the 300 mg/kg/day group (14/15).

Relative liver weights were incr eased (p<0.05) by 8% at 150 mg/ kg/day, and absolute, adjusted and relative liver weights were increased (p< 0.05) by 9 to 14% at 300 mg/kg/day. Absolute, adjusted and relative adrenal weights were all increased (p<0.05) by 24% at 150 mg/kg/day, and by 36 to 42% at 300 mg/kg/day. Absolute and adju sted, but not relative, pituitary weights were decreased (p<0.05) by 12% at 300 mg/kg/day. Kidney, ovary and t hyroid weights were unaffected by treatment.

Serum T 4 concentrations were redu ced at 150 and 300 mg/kg/day ( ↓ 97% and ↓ 93%, respectively; p<0.0001); however, TSH levels were unaffected by treatment. The repo rt states that T 4 and TSH were conducted using radioimmunoassays; no fu rther information is provided on the exact method used, or on method validation a nd quality control procedures.

Iprodione was not associated with any changes in thyroid follicular cell height or colloid area. There were no effects of treatment on the ovary or kidney. In the uterus, iprodione treatment
was associated with minimal to mild decrea ses in uterine horn diameter characterized by decreased stromal development in 2/16 animals at 150 mg/kg/day and in 5/16 animals at 300 mg/kg/day.

The doses tested were consider ed adequate based the results of range-finding and uterotrophic studies, and on the estrogen-related effects seen at the high dose in this assay.IPRODIONE FINAL

Page 41 of 66 Male Pubertal Assay (OCSPP 890.1500)

The requirement for a Tier 1 AR bindi ng assay was satisfied by OSRI: Blystone et al. (2007) and results from the female pubertal assay (for t hyroid parameters). Results from the Blystone et al. 2007 study are presented in Appendix 2.

Steroidogenesis Assay (OCSPP 890.1550)

The requirement for a Tier 1 steroidogenesis as say was satisfied by OSRI (Blystone et al., 2007). The results from this study are pres ented with the OSRI in Appendix 2.

Uterotrophic Assay (OCSPP 890.1600)

In a Uterotrophic Assay (MRID 48617205) , iprodione (98.1% a.i., lot # AE F062470-01-08) in aqueous 0.4% methylcellulose was administered daily via oral gavage to groups of eight immature female Sprague Dawley rats at dose levels of 0 (vehicle), 200, or 400 mg/kg/day on post-natal days (PND) 19-21. A positive contro l group was treated with a daily dose of 17 α

ethynyl estradiol (EE) at 1 μ g/kg/day in corn oil by oral gava ge. Body weights were determined daily. All animals were terminated on P ND 22 approximately 24 hours after the final dose
administration to determine wet and blotted uterine weights.

No adverse, treatment-related effects were observe d on mortality or clinical signs. Body weights were decreased (p<0.05) on Day 4 ( ↓ 12%) in the high dose group compared to the vehicle controls, resulting in a decrease (p<0.05) in ove rall body weight gain ( ↓ 54%). This finding
indicates that the dose levels select ed for the study were adequate.

No abnormal gross observations were seen in the uterus. Uterine weights in the iprodione treated groups were comparable to the vehicle controls. Absolute wet and blotted uterine weights for the positive control (EE) group were each increased (p<0.05) by 270%.

No statistically significant changes were seen in uterine weight in this assay. Iprodione was negative in the uterotrophic assay.IPRODIONE FINAL

Page 42 of 66 APPENDIX 2. Other Scientifically Relevant Information (OSRI)

Published literature and Part 158 studies consid ered relevant for dete rmining the potential interaction of iprodione with the endocrine syst em are summarized below. Given the breath of toxicity data available, the onl y Part 158 studies discussed in th is appendix are those that fulfill current guideline requirements and are directly relevant to asse ssing potential endocrine effects ( i.e. reproductive and developmental studies, subc hronic and chronic toxi city studies, special mechanistic studies, etc.).

AR Binding Assays

In an AR binding assay, Fail et al . 1994 examined the affinity of iprodione and seven of its metabolites (RP32490, RP255040, RP36112, RP36114, RP36115, RP36118, RP36119) to the AR binding site. The methods utilized for this assa y were similar to the methods utilized in the current Guideline AR binding assay. Ventral prostate cytosol from intact male SD rats was used
as the source of AR to conduct both Saturation and Competitive Binding Experiments. Separate Competitive Binding assays were conducted to meas ure the binding of a single concentration of [ 3 H]-R1881 (1 nM) in the presen ce of increasing concentrat ions (1, 5, 10, 50, 100, 500, and 1000

μ M) of iprodione (97.3% purity) and each of seven metabolites. The solvent vehicles used in the study were acetonitrile for iprodione and five me tabolites, dimethylformamide for the two other metabolites, and ethanol for the remaining referen ce standards. Final assay concentrations of the solvent vehicles were not reported, but the assa ys were tested for possible solvent vehicle interference. The assay included DHT, testosterone, flutamide, and hydroxyflutamide as positive controls, and R1881 as the ligand reference standard.

Saturation Binding data were provided in the st udy report. The mean di ssociation constant (K d ) for [ 3 H]-R1881 was 4.33 nM, and the B max was 135 fmol/mg protein for prostate cytosol. In the Competitive Binding Experiment, iprodione inhibited [ 3 H]-R1881 binding by <15% at the highest concentration tested, 1,000,000 nM (10 -3 M); therefore, an IC 50 and RBA could not be calculated for iprodione. However, one of its me tabolites (RP25040) had an affinity for the AR binding site similar to fl utamide. The calculated IC 50 values were as follows: R1881, ≈ -8.5 log M; DHT, -8.5 log M; testosterone , -8.0 log M; flutamide, -4.5 log M; hydroxyflutamide, -5.7 log M; metabolite RP25040, -4.1 log M; metabolite RP36112, -3.8 log M; and metabolite RP36115, -3.5 log M. The corresponding RBA values we re R1881, 100%; DHT, 100%; testosterone, 35%; flutamide, 0.013%, hydroxyflutamide, 0.16%; metabolite RP25040, 0.0064%; metabolite RP36112, 0.0028%; and metabolite RP36115, 0.0012%. Based on these data, iprodione is a nonbinder in this test system.

In another competitive AR binding assay, Blystone et al . (2009 ) investigated the binding potential of iprodione to the human androgen receptor (hAR) e xpressed within COS cells. COS-1 cells were transiently transfected with 0.5 μg of pCMV5-hAR, and were exposed the followingIPRODIONE FINAL

Page 43 of 66 day to iprodione at 1 to 200 μM (n=3) or vinc lozolin at 0.01 to 10 μM (n=2) to evaluate the ability of the test substances to displace radiolabeled [ 3 H]-R1881 at 0.5 nM (specific activity 83.5 Ci/mM) from the hAR; a 100-fold molar ex cess of non-radiolabeled R1881 was added at
each concentration of the test substances to de termine nonspecific binding. After incubation for 2 hours, the cells were washed with phosphate buffered saline to remove unbound ligand, and the cells were lysed. The cell debris was collected fo r measurements via liquid scintillation counting. The specific binding for each test substance wa s determined by subtracting nonspecific binding
counts for unlabeled R1881 from the total bi nding counts at each concentration, and was expressed as percentage bound (ratio of specifi c radioligand bound at each concentration vs. radioligand bound in the absence of the chemical competitor × 100).

Iprodione bound to the hAR with an IC 50 of 86.0 μM compared to an IC 50 of 1.4 μM for vinclozolin, suggesting that iprodione or one of its metabolites is a weak AR antagonist in this
test system.

AR transcriptional Activation Assays (ARTA)

In a published, non-Guideline study (Kojima et al ., 2004), the potential of 200 chemicals (including iprodione; purity 95-100%) to interact with two human ER subtypes (hER α and
hER β ) and one human androgen receptor (hAR) in tr ansactivation assays with Chinese hamster ovary cells (CHO-K1 cells) was inve stigated. For detection of hER α or hER β activity, cells were transfected with pcDNAER α or pcDNAER β , pGL3-tkERE, and pRL-SV40. For detection of hAR activity, cells were transfected with pZ eoSV2AR, pIND-ARE, an d pRL-SV40. Flutolanil was tested over a concentration range from 10

− 8 to 10

− 5 M, and the responses were compared to the activity of 17 β -estradiol (E 2 ; purity >97%) or 5 α -dihydrotestosterone (DHT; purity 95%) for the ER or AR assays, respectively. For meas urement of antagonistic activity to hER α , hER β and hAR, either 10

− 11 or 10

− 10 M E 2 or 10

− 10 M DHT was added to the cell cultures along with the test compound, respectively. A gonistic activities were eval uated by the relative activity expressed as 20% relative effective concentration (REC 20 ), the concentration of the compound demonstrating 20% of the activity of 10

− 10 M E 2 , 10

− 9 M E 2 , or 10

− 9 M DHT for ER α , ER β , and AR, respectively. Antagonistic activities were evaluated similarly by the relative activity expressed as 20% relative i nhibitory concentration (RIC 20 ), the concentrat ion of the compound showing 20% inhibition of the activity of E 2 or DHT against ER α , ER β , and AR, respectively. Iprodione did not show agonist or antagonist activity for ER α , ER β , or AR in this study.

Blystone et al. (2007 ) investigated the possibl e AR agonist or antagonist activity of iprodione in

vitro using a stably-transfected MMTV luciferase reporter MDAkb2 cell line. Luciferase reporter cells (1 x 10 -4 /well, 100 μL/well) were passed into a 96-well luminometer plate in L-15 media supplemented with 10% fetal bovine serum, 100 U/mL penicillin, 100 μg/mL streptomycin, andIPRODIONE FINAL

Page 44 of 66 0.25 μg/mL amphotericin B. Cells were allowed to attach for 4–6 hours, then exposed for ~20 hours to 100 pM to 10 μM iprodione in ethanol ( ≤ 0.1% v:v), either alone or in combination with 0.1 μM DHT, in quadruplicate. The test included background control wells containing media alone, and positive controls of 0.1 μM DHT and 0.1 μM DHT + 0.1 μM flutamide (AR antagonist). Luciferase activity was measured w ith a luminometer; induction was calculated as fold increase over background.

The results of the AR-related tran scriptional activation assay indi cated that iprodione at 100 pM

to 10 μ M did not increase luminescence over background, and that the positive DHT response was not reduced by treatment with iprodione at th e concentrations tested. Cytotoxicity was not apparent at up to 10 μ M iprodione.

In a related study (Blystone et al., 2009), iprodione was tested in the same in vitro assay under
the same conditions at concentrations from 1 to 300 μM, either alone or in combination with 0.1 nM DHT, in quadruplicate (4× wells/dose, n=8) . This test also incl uded background control wells containing media alone, and positive controls of 0.1 μM DHT and 0.1 μM DHT + 0.1 μM flutamide. Luciferase activity was measured with a luminometer; inducti on was calculated as fold increase over background.

In this study, iprodione alone did not increase luminescen ce over background; however, a significant increase in cytotoxic ity was observed at 300 μM iprodi one. Treatment with iprodione
+ DHT resulted in a decrease in DHT fold-inducti on with increasing iprodione concentrations (IC 50 =245.9 μM), suggesting that iprodi one may act as an AR antagonist.

Hershberger Assay

In a Hershberger assay conducted by Blystone et al . (2009), castrated male rats were exposed to iprodione in conjunction with testosterone. Immature SD ra ts, castrated on PND 42, were maintained under standard laboratory conditi ons with access to rat chow and tap water ad
libitum . Prior to dosing, animals were ranked by we ight and assigned to treatment groups (6 rats/group) by weight to minimize differences in means and variance. Starting on PND 49, the rats received a daily subcutaneous (s.c.) injection of 100 μg test osterone proprionate (TP) in 0.1
mL corn oil and were dosed da ily by oral gavage with iprodi one at 50, 100, or 200 mg/kg in a dose volume of 2.5 mL/kg bw; a control group recei ved s.c. injections of TP and was dosed orally with corn oil. After 10 to 11 days of treatment, animals were euthanized and necropsied. Terminal body weights were recorded, and organ weights were recorded for the liver, paired kidneys, paired adrenal glands, glans penis, ventral prostate, seminal vesicle, LABC, and Cowper’s gland. Blood was collected by cardi ac puncture for serum measurements of
testosterone and lutein izing hormone (LH). Hormones were an alyzed by radioassay using eitherIPRODIONE FINAL

Page 45 of 66 published procedures (LH) or a commercially ava ilable kit (testosterone); the reported detection limit was 0.04 ng/mL for testosterone.

In this Hershberger assay, th ere were no significant treatme nt-related effects observed for iprodione on terminal body weight or body weight gain; weight of kidneys, glans penis, or Cowper’s gland; or serum testosterone or LH concentrations. Iprodione significantly decreased androgen-sensitive organ weights for the ventral prostate at 200 mg/kg ( ↓ 30%, p<0.01) and for the LABC at all doses ( ↓ 10 to 11% at 50 and 100 mg/kg, p<0.05; ↓ 21% at 200 mg/kg, p<0.001); a significant (p<0.05) decreasing trend was also observed for the seminal vesicle weights. Significant increases were observed in liver weight at 100 to 200 mg/kg ( ↑ 6 to 10%; p<0.05) and in adrenal weights at 200 mg/kg ( ↑ 44%, p<0.001). Based on the signi ficant decreases observed in three androgen dependent tissues, iprodione wa s positive for anti-androgenic activity in this assay. The authors also noted that the absence of an effect on serum testosterone suggested that iprodione does not reduce circulating te stosterone by increasing its metabolism.

Male Pubertal Rat Study

In a study investigating the effect of iprodione on test osterone production and sexual development in male rats (Blystone et al., 2007), immature male SD rats (8 rats/group) were dosed orally once a day with iprodione (97% purity) in corn oil at 0, 50, 100 or 200 mg/kg/day from PND 23 to 51/52. Body weights were reco rded at study initiation, PND 41, and at PPS initiation and completion. The progression of PPS was inspected daily from PND 37 to necropsy. On the final day of dosing, animals we re necropsied after deca pitation within 1.25 to 3.50 hours of the final dose. Terminal body weight s were recorded, and organ weights were recorded for the liver, paired kidneys, paired adrenal glands, glans penis, ventral prostate, seminal vesicle, LABC, and paired Cowper’s glands, testes, and epididymides. Trunk blood
was collected for serum measurements of testosterone, progesterone, 17 α

hydroxyprogesterone, androstenedione, estradiol, aldosterone, corticosterone, and LH. Hormones were analyzed by radioassay with either published procedures (for LH) or commercially available kits (remaining hormone s); reported detection limits were 0.04 ng/mL for testosterone and androstenedione, 0.02 ng/mL for progesterone, 0.07 ng/mL for

17 α -hydroxyprogesterone, 11 pg/mL for aldoster one, 5.7 ng/mL for corticosterone, and 0.115 ng/mL for LH.

There were no significant eff ects of iprodione on body weight or body weight gain, nor was systemic toxicity observed at up to 200 mg/kg/ day of iprodione. However, iprodione delayed (p<0.05) the initiation and completion of PPS at 100 and 200 mg/kg vs. control, with an increase (p<0.05) in body weight at PPS: PPS initiation and completion were delayed by 2 days at 100 mg/kg and by 2.6 to 2.8 days at 200 mg/kg, a nd body weight at initiation and completion of PPS was increased by 9 to 10% at 100 mg/kg a nd by 16% at 200 mg/kg. At necropsy, iprodioneIPRODIONE FINAL

Page 46 of 66 at 200 mg/kg/day increased (p<0.01) liver and adrenal weights by 14% and 40%, respectively, and decreased (p<0.05) seminal vesicle a nd epididymides weights by 24% and 11%, respectively. There were no signi ficant treatment-related effects on the weights of the kidneys, glans penis, ventral prostate, LA BC, Cowper’s glands or testes.

There was no treatment-related effect on seru m LH concentrations, but serum testosterone concentrations were decreased (p<0.0 001) by 73%, 83%, and 89% at 50, 100, and 200 mg/kg/day, respectively. A decreasin g progesterone trend (NS) was also observed at all doses.
Results for 17 α -hydroxy progesterone, andros tenedione, aldosterone, a nd corticosterone were considered to be equivocal because many of the values for these hormones were below the detection limits. Based on non-parametric analys es via the Kruskal-Wallis test for these hormones, serum 17 α -hydroxyprogesterone and androstenedione concentrations were decreased (p<0.05) at 100 and 200 mg/kg, serum aldostero ne was decreased (p<0.05) at 50 and 200 mg/kg, and there was no treatment effect on corticosterone concentrations.

Iprodione effects on serum test osterone concentrations (LOAEL 50 mg/kg/day) were more sensitive than iprodione eff ects on PPS (LOAEL 100 mg/kg/day) or androgen-dependent organ weights (LOAEL 200 mg/kg/day).

The study demonstrated that iprodione delays male rat pubertal development and inhibits testicular steroidogenesis. The authors noted th at the decreases observe d in testosterone, along with the decreasing trends observed for progest erone suggested a block in steroidogenesis at CYP11A (P450scc; cholesterol side-chain cl eavage enzyme), StAR (steroidogenic acute regulatory protein), or a precedi ng event. Because serum LH concentrations were not reduced, effects on the hypothalamic-pituitary axis were deemed unlikely.

Steroidogenesis Assay

The effect of iprodione on te sticular steroidogenesis were investigated by Blystone et al. (2007)
in an ex vivo assay using testicular tissue from immatu re male rats. Immature male SD rats (8 rats/group) were dosed orally on ce a day with iprodione (97% purit y) in corn oil at 0, 50, 100 or 200 mg/kg/day from PND 23 to 51/52. At sacrifice, the right testis of each test animal was removed, de-capsulated, weighed, and sectioned in to 50- to 100-mg pieces. Two of the pieces from each rat were incubated in oxygenated Gibco M199 media (95% O 2 ; 5% CO 2 ) with 100 mIU/mL hCG, and two pieces were incubated in media without hCG. Testis sections were incubated for 1.5 hours at 34oC, after which the me dia was collected by cent rifuge and stored at -70oC for quantification of testosterone, progesterone, 17 α -hydroxyprogesterone, and
androstenedione. Hormone concentrations in th e media were analyzed by radioassay with commercially available kits. The reported detec tion limits were 0.04 ng/mL for testosterone and androstenedione, 0.02 ng/mL for pr ogesterone, and 0.07 ng/mL for 17 α -hydroxyprogesterone.IPRODIONE FINAL

Page 47 of 66 In the ex vivo testing, testosterone concentrations were decrease d (p<0.05) with increasing iprodione concentrations, and th e reduction in testosterone co ncentrations were similar under stimulated (hCG present) and unstimulated condi tions. No significant treatment-related effects
were observed on concentrations of 17 α -hydroxyprogesterone or andros tenedione (stimulated or unstimulated); however, there was a trend (NS) in decreased 17 α -hydroxyprogesterone concentrations with increasing iprodione con centrations. There was also a trend (NS) in decreased progesterone concentr ations with increasing iprodione concentrations, with the reduction becoming significant (p<0.05) at 200 mg/kg of iprodione under the stimulated
incubations.

The authors noted that the decreases observe d in testosterone c oncentration in the ex vivo testing, along with the decreasing trends observed for pr ogesterone concentration suggested a block in steroidogenesis at CYP11A (P450scc; choleste rol side-chain cleav age enzyme), StAR (steroidogenic acute regulatory protein), or a preceding event. Direct inhibition of steroidogenesis by iprodione was further supporte d by the finding that hCG stimulation was unable to overcome the inhibitory effect of iprodione on testosterone production.

90-Day (Subchronic) Oral T oxicity in Rodents (Rat)

In a subchronic rat feeding study (MRID 42960701), 10 Crl:CD(SD)BR ra ts/sex/group were administered iprodione (95.7% purity) in th e diet at nominal doses of 0, 1000, 2000, 3000, and 5000 ppm for 90 days. These nominal dietary levels are respectivel y equivalent to doses of 0, 78, 151, 252, and 355 mg/kg/day for males and 8 9, 189, 266, and 408 mg/kg/day for females. Clinical signs of toxicity included hunched pos ture, piloerection, pale and/or cold extremities and an emaciated appearance. The 5000 ppm dose group was terminated early (Week 8) due to excessive systemic toxicity (body weight loss, decreased food consumpti on, and clinical signs).

Body weights for the medium (2000), high (3000 ), and 5000 ppm dose groups were decreased for the males by 25, 48 and 61%, respectively, and for females by 24, 30 and 55%. The 5000 ppm group had body weight losses for both sexes, and the body weight gains were decreased at the medium and high doses by 39 and 74% for males and 30 and 62% for females. Food consumption was also decreased by 19% for me dium-dose males and by 31 and 21% for males and females at the high dose; food efficiency wa s also decreased for both sexes at the medium and high doses. Based on the substa ntial reductions in body weight ( ≥ 24%) and body weight gains ( ≥ 30%) at the medium and high doses, these doses showed overt toxicity.

The sex organs, pituitary, and adrenals of both sexe s appear to be target organs for iprodione. In general, the decreases observed in organ weights and the acco mpanying increases in relative organ weights may be attributed to the decrease d body weight, but in females, decreased relativeIPRODIONE FINAL

Page 48 of 66 organ weights were observed in th e uterus, ovary, adrenal, and pitu itary, mainly at the high dose. Absolute pituitary weight was decreased (p< 0.05) in males by 20 and 35% at the medium and high doses, respectively, and in females (NS) by 15, 24 and 48% at the low, medium and high doses, respectively, with a significant decrease (p<0.05) in females in the relative weight ( ↓ 26%) at the high dose. Absolute thyroid weight was decreased (p<0.01) by 42% in males at the high dose. In females, absolute uterus weight was decreased (p<0.01) by 74% at the high dose, and absolute ovary weight was decreased (p<0.01) by 21 and 51% in the mid- and high-dose groups. Relative liver weights at the medium and high doses were increased (NS) by 15 and 32% in males, and 26 and 29% in females. No treatmen t-related changes were seen in absolute or relative weights of the testes or adrenal glands at any dose.

Gross examinations at necropsy id entified the following findings: • Testes were small in 1/10 at the high dose versus 0/10 in the control;
• Seminal vesicles were small with minimal conten ts in 6/10 at the high dose versus 0/10 in the control; • Prostate was small in 4/10 at the high dose compared to 0/10 in the control; • Uterus was thin at the medium (5/10) and hi gh (10/10) dose groups compared to 0/10 in the control; and • Adrenals were pale in 6/10 males at the high dose and in females in the medium (5/10) and

high (10/10) dose groups compared to 0/10 in the control.

Histopathological examinations identified the following findings: • Testes with atrophy of the seminiferous tubu les in 6/10 at the high dose versus 0/10 in the

control; • Epididymides with abnormal spermatozoa (6/10) and absence of spermatozoa (5/10) compared

to none in the control;
• Seminal vesicles with reduced secretions at the mid (9/10) and high (7/10) dose compared to

0/10 in control; • Prostate was atrophied at the mid (5/10) and hi gh (9/10) dose compared to none in the control; • Uterus was atrophied at the mid (2/10) and high (9/10) dose compared to none in the control; • Ovaries had reduced number of corpora lutea at the mid (2/10) and high (4/10) dose compared
to none in the controls; and • Adrenals had enlargement of the cells of the zona glomerulos a in both sexes at the medium

dose (9/10 males, 9/10 females) and high dose (9/10 males and 10/10 females) compared to
none in the control. Fine vacuol ation of the cells of the zona fasciculata was also noted in males at the mid- (3/10) and high- (9/10) dose groups compared to none in the control.

The NOEL is 1000 ppm (male 78/female 89 mg/ kg/day), and the LOAEL is 2000 ppm (male 151/female 184 mg/kg/day), based on d ecreased body weight/g ain, decreased food consumption/ food utilization, organ weight effect s, and microscopic lesions in the sex organs.IPRODIONE FINAL

Page 49 of 66 90-Day (Subchronic) Oral Toxi city in Rodents (Mouse)

In a 90-day oral toxicity study (MRID 48125502), iprodione (95.7% pur ity) was administered in the diet to 10 Crl:CD-1 (I CR) BR mice/sex/treatment group at nominal doses of 0, 1500, 3000, 6000, or 12000 ppm (equivalent to 0, 260/ 327, 509/659, 1066/1303, or 2130/2608 mg/kg/day for males/females) for 13 weeks. No effects of treat ment were observed in body weight gain or food consumption.

Dark and/or swollen abdomens were observed in the mid-low-, mid-high- and high-dose groups, with the effect increasing in frequency with incr easing dose, and occurring as early as Week 1 in the high-dose group. Other clinic al signs of toxicity observed in both sexes in the high-dose group included lethargy, pilo erection and hunched posture.

Adjusted (for body weight) liver weights were do se-dependently increased (p<0.01) in males at the mid-low ( ↑ 26%), mid-high ( ↑ 98%) and high dose ( ↑ 133%) and in females at the low ( ↑ 19%) mid-low ( ↑ 41%), mid-high ( ↑ 98%) and high dose ( ↑ 149%). Absolute adrenal weights were also dose-dependently increased (p<0.01) in males at the mid-high ( ↑ 28%) and high dose ( ↑ 32%), and in females at the low ( ↑ 28%) mid-low ( ↑ 31%), mid-high ( ↑ 48%) and high dose ( ↑ 30%). Also at the high-dose group, adjusted kidney weights were decreased (males, ↓ 31%; females, ↓ 17%) and absolute spleen wei ghts were increased (males, ↑ 15%; females ↑ 45%). Decreases
were also noted in absolute uterus wei ght in the mid-high- and high-dose groups ( ↓ 29, p<0.05 and 58%, p<0.01) and in absolute and adjusted ovary weights ( ↓ 50–53%, p<0.01) in the high-dose group. No treatment-related changes were se en in absolute or adjusted weights of the testes, pituitary or thyroid glands at any dose.

Grossly, the liver was enlarged with irregular su rface depressions in most animals in the midhigh- and high-dose groups. Other endocrine-related hi stological findings included: small uteri at the mid-high (4/10) and high (7/9) dose compared to 0/10 in control; ut erine atrophy at the midhigh (4/10) and high (7/9) dose; and the absence of corpora lutea in the ovaries at the mid-high (2/10) and high (8/9) dose. In th e adrenals, there was increased in cidence of enlargement of the cells of the zona glomerulosa in males (8/10) at the high dose and in females at the mid-high (6/10) and high (8/8) dose; enlargement of the zona fasciculata cells in m ales (3/8) at the high dose, and vacuolation of these cells was observe d in males (2/8) and fe males (4/9) at the high dose, as well as in females from the low (1 /10), mid-low (3/10), a nd mid-high (6/10) dose groups. No treatment-related histopa thological changes were seen in the testes, epididymides, seminal vesicle, prostate, vagina, mammary glands, or thyroid and pituitary glands.

The LOAEL is 3000 ppm (males, 509/ females 659 mg/kg/day), based on clinical signs of toxicity (abdomen dark and/or swollen), gross pathological fi ndings in the liver, increasedIPRODIONE FINAL

Page 50 of 66 adrenal weight, and microscopic adrenal findings in the females, and incr eased liver weights and microscopic liver findings in both sexes. The NOAEL is 1500 ppm (males 260/ females 327 mg/kg/day).

Prenatal Development in Rodents (Rat)

In a developmental toxicity study (MRIDs 00162984 and 40514901), 20 pregnant SD CD rats (mated 1:1) were administered iprodione (94.2% purity) by gavage in 0.5% methylcellulose at doses of 0, 40, 90, and200 mg/kg/day from gestat ion day (GD) 6 through 15. On GD 20, the
dams were sacrificed via CO 2 inhalation. There were no deaths . Body weights were comparable among the groups. There were no significant differen ces observed in the mean number of viable fetuses, implantations, corpora lutea, and resorp tions, and pre- and post- implantation losses were comparable among the groups. There was no evidence of maternal toxicity at any dose (maternal NOAEL >200 mg/kg/day). The developmen tal NOAEL was 90 mg/kg/day, and the developmental toxicity LOAEL was 200 mg/kg/ day, based on delayed fetal development (slightly reduced fetal body weight and increased incidences of space between the body wall and organs in the fetuses).

In a special developmental toxicity study (MRID 44365001) examining sexual differentiation, pregnant SD rats (25/dose) recei ved iprodione (97.1% purity) in methylcellulose via gavage at doses of 0, 20, 120, or 250 mg/kg/day during GD 6 through 19. The study included a positive control group that rece ived flutamide at 50 mg/kg/day. For maternal toxicity, the NOAEL is 20 mg/kg/day, the LOAEL is 120 mg/kg/day, based on decreased body-weight gain and decreased food efficiency. At 250 mg/kg/day, deaths occurred (9/ 25) in addition to decreased body-weight gain and food consump tion/efficiency. No treatment -related adverse effects were seen in pregnancy rate, the numbe rs of corpora lutea, implantati ons, and live fetuses per litter. There was no significant increase in the number of resorptions per litter at any dose, but the highdose group displayed an increase in late resorptions per litter co mpared to the control (0.47 vs. 0.09), and post-implantation loss (13.5%) was ap proximately double that of the vehicle control (6.8%). There was no increase in the incidence of external malformations with increasing dose of iprodione, but there was an increase in the number of runts at the high dose. In males, dosedependent decreases were seen in anoge nital distance (AGD) in the mid- (p<0.05, ↓ 5%) and high- (p<0.01, ↓ 14%) dose groups. In females, the AGD index (AGD/body weight) was increased (p<0.01) at the high dose. For developmental toxic ity, the NOAEL is 20 mg/kg/day and the LOEL is 120 mg/kg/day, based on decreased AGD in the male pups.

In a published development toxicity study by Wolf et al. (1999), eight pregnant SD females were dosed orally with iprodione ( 99% purity) at 100 mg/kg/day from GD 14 to Day 3 of lactation. Doses were adjusted daily based on individual mate rnal weight changes. At about 5 months of age, F1 males (number not specified) were sa crificed by decapitation after removal from theIPRODIONE FINAL

Page 51 of 66 home cage in a separate room during the dark phas e of the animal’s daily cycle. Each male was weighed and their ventral surface was shaved a nd examined for abnormalities, including the number and location of retained nipples, cleft phallus, vaginal pouch, and hypospadias. The animals were also examined internally for ectopic or atrophic test es; agenesis of the gubernaculum, epididymides, sex accessory gl ands, and ventral prostate; epididymal granulomas; hydronephrosis; and enlarged bladde r with stones. The following tissues were collected and weighed: pituitary, adrenal, kidneys, liver, ventral prostate, seminal vesicle with coagulating gland and fluid, tes tis, and epididymis. Testicular, epididymal, and sex accessory tissues were taken histological evaluation. The study authors re ported that iprodione did not demasculinize or feminize male pups, but actual data from the control and iprodione-treated animals were not provided.

Prenatal Development in Non-Rodents (Rabbit)

In a developmental toxicity study (MRID 00155469) , artificially-inseminated New Zealand female rabbits (18/dose group) were admini stered iprodione (95.0 to99.3% purity) in 0.5% aqueous methylcellulose at doses of 0, 20, 60, and 200 mg/kg/day via gavage from GD 6 through 18. On GD 29, the does were sacrificed. Seven high-dose does aborted between GD 17 and 23, and prior to aborting all had displayed decrea sed urination and defecation. One mid-dose doe (GD 28) and one control doe (GD 20) also aborted. All other does survived until study termination, and nine of the high-dose does that did not abort displayed decreased urination and defecation. During the dosing period, the mid-dose doe s gained less weight than control, and the high-dose does lost weight. A negative net bodywe ight gain was observed at the mid- and highdoses. The high-dose does displayed decrease d food consumption during the dosing period. Gravid uterine weight was decreased at the high-dose (90% of control). No treatment-related changes were seen in pregnancy rate, numb er of corpora lutea, mean number of implantations/litter, post implantation loss, early or late resorptions, number of live fetuses/litter,
or soft tissue abnormalities. The maternal NOA EL was 20 mg/kg/day, and the maternal LOAEL was 60 mg/kg/day, based on decreased body-weight gain. At the highest dose tested (200 mg/kg/day), maternal toxicity was demonstrated by an increased rate of abortions (7 does), bodyweight loss, decreased food consumption, and decr eased defecation and urination in females that
aborted.

The developmental toxicity NOAEL was 60 mg/kg/ day, and the developmental toxicity LOAEL was 200 mg/kg/day, based on an increased in cidence of skeletal variations (13 th full rib, malaligned sternebrae, and/or 27 pr esacral vertebrae, with or without delayed ossification).

Two-Generation Reproduction in Rodents (Rat)IPRODIONE FINAL

Page 52 of 66 In a 2-generation reproducti on study (MRIDs 00162983 and 41871601), 28
Crl:CD ® BR/VAF/PLUS rats/sex/group were administered iprodione (9 6.2% purity) via the diet at nominal doses of 0, 300, 1000, and 3000/2000 pp m for two generations (2 litters per generation). These doses were equivalent to 0, 18.5, 61.4 and 154.8 mg/kg/day in males, and 0, 22.5, 76.2 and 201.2 mg/kg/day in females. The F0 animals were fed the test diets for 70 days and were mated to produce the F1a offspring. Afte r a 2-week resting period the F0 were mated again to produce the second litter (F1b). The F1 animals were mated twice in a similar manner after 70 days of treatment to produce the F2a an d F2b generations. Organ weight data were not provided, but histopathologi cal examinations were conducted on selected tissues.

No treatment-related effects were observed in ma le or female mating, fertility, gestation indices, gestation length, and fetal sex ra tio in any generation. However, offspring toxicity observed in the F1a generation included a d ecrease (p<0.05) in the number of live pups/litter at the high dose (12.0) compared to control (13.7) on Day 0 and a d ecrease (p<0.01) in the viability index at the high dose (96%) compared to controls (99%). Toxicity observed at the high dose in the F1b generation included: a decrease (p<0.01) in the number of liv e pups/litter on Day 0 (9.4 vs. 13.7 control) and Day 4 pre-cull (8.7 vs. 13.4 control); a decrease (p <0.01) in the viability index (79%) compared to control (98%); and a decr ease (p<0.01) in the lactation index (67%) compared to control (98%). In addition, the numbe r of live pups/litter wa s decreased at the high dose in both the F2a generation (p<0.01; 11.9 vs. 14.2 control on Day 0) and the F2b generation (p<0.05; 11.7 vs. 14.6 control on Day 0). With re gards to reproductive tissues, no treatmentrelated histopathological lesions we re seen in the testes, epididym ides, seminal vesicle, prostate, coagulating gland, ovaries, uterus, vagina, cervix, or the pituitary gland in the F0 and F1 parental
animals.

The systemic maternal/parental NOAEL was 30 0 ppm (male, 18.5/female 22.5 mg/kg/day), and the LOAEL was 1000 ppm (male 61.4/female 76.2 mg /kg/day), based on decreased body weight, body-weight gain, and food consumption in both sexes and both generations. The reproductive (offspring) NOAEL was 1000 ppm (76.2 mg/kg/day) , and the reproductive (offspring) LOAEL was 2000 ppm (201.2 mg/kg/day), based on decreased pup viability (as evidenced by an increased number of stillborn pups and decrease d survival during PND 0 to 4), decreased pup body weight throughout lactation, and an increased incidence in clinical signs in pups during the lactation period (smallness, reduced mobility, unkempt appearance, hunching, and/or tremors).IPRODIONE FINAL

Page 53 of 66 Chronic Oral Toxicity/Carcinogenicity in Rodents (Rat)

In the combined chronic t oxicity/carcinogenicity study in rats (MRIDs 42637801 and 42787001), iprodione (95% purity) was admi nistered to 60 SD rats/sex/dos e group via the diet at nominal doses of 0, 150, 300, and 1600 ppm (males: 6.1, 12.4, and 69 mg/kg/day; females: 8.4, 16.5, and 95 mg/kg/day, respectively) for 24 months. An additional 10 rats/sex/group were administered iprodione for 52 weeks (interim sacrifice).

There were no adverse effects on survival or clini cal signs in either sex. Body-weight gains were decreased in both sexes at the high-dose compar ed to control. Overall, body-weight gains decreased by 14% and 8% in the high-dose male s and females, respectively; and body weight gains at Week 12 were decreased by 14% in hi gh-dose males and by 19% in high-dose females. Food consumption was also decreased slightly at th is dose in both sexes. At the interim sacrifice (Week 52), there were no treatment-related clinical pathology findings in e ither sex or changes in organ weights. At Week 52, high-dose males displaye d an increase in the incidence of lesions in the adrenals, and there was an increased incidence of centrilobular hepatocyte enlargement in mid- and high-dose males. High-dose females displa yed an increase in cent rilobular hepatocyte enlargement and an increase in the incidence of generalized rarefaction a nd fine vacuolation of the zone fasciculata in the adrenals comp ared to the control and other dose groups.

No treatment-related changes were seen in absolute or relative weights of the ovaries, uterus, adrenals, or pituitary glands at terminal sacr ifice in any dose group. However, adjusted liver weight was increased by 9% (p<0.05) and 11% (p<0.01) in mid- and high-dose males, respectively. In the high-dose males, adjusted and relative weights of the testes (with epididymides) were also increase by 17% (p <0.05) and 30% (p<0.01), respectively, and the adjusted thyroid weight was increased by 14% (p<0.05) compared to control.

At termination, in males at the high dose, increas es (p<0.01) were observed in the incidences of testicular masses (37% at the hi gh dose vs. 0% in the control), small seminal vesicles (18% vs. 5%) and small prostate (8% vs. 0%).

No treatment-related histopathological lesions were seen in the ut erus, and mammar y, thyroid or pituitary glands at any dose at terminal sacrif ice. However, a number of histological changes were noted in the reproductiv e tissues of high-dose males:

 increase (p<0.01) in the inci dence of interstitial cell hype rplasia in the testes (58%) compared to control (12%);

 increase (p<0.01) in the incide nce of atrophy of the seminife rous tubules (40%) compared to control (17%);

 increase (p<0.05) in reduced spermatozoa (13% ) in the epididymides compared to control (0%);IPRODIONE FINAL

Page 54 of 66  increase (p<0.01) in the incide nce of atrophy of the prostate (22%) compared to control (3%);

 increase (p<0.05) in the promin ent abnormal spermatogenic cells (8%) compared to control (0%); and

 reduction (p<0.01) in secretion (20%) in semi nal vesicles compared to control (7%).

In addition, there was an increase (p<0.01) in the incidence of un ilateral interstiti al cell tumors (25%) and bilateral interstitial cell tumors (23 %) in high-dose males compared to control (5% and 0%, respectively), and thes e incidences exceeded the hi storical range (0 to10%).

Other histological findings includ ed an increase (NS) in the inci dence of tubular hyperplasia in the ovaries (17%) of high-dose females compared to control (7%), and an increase (p<0.05) in centrilobular hepatocyte enlargement (10%) in hi gh-dose males compared to 0% in control. In addition, in the adrenals, there were increased (p<0.01) incidences in the following lesions: (i) focal enlargement of cells of the zona glomerul osa (mid- and high-dose males and females); (ii) generalized enlargement of cells of the zona glomerulosa (high-dos e males); and (iii) rarefaction
and vacuolation of the zona fasciculata and fine vacuolation of the zona reticularis in the midand high-dose males.

The NOAEL for non-neoplastic changes is 150 ppm (males, 6.1 mg/kg/day; females 8.4 mg/kg/day), and the LOAEL is 300 ppm (males 12.4 mg/kg/day; females 16.5 mg/kg/day), based on histological changes in the adrenals of ma les and females, an increased incidence of interstitial cell hyperplasi a, reduced spermatozoa in the epid idymides, reduced secretion of the seminal vesicles, increased hemosiderosis in the spleen in females, and increased liver weight.
There was an increase in the incidence of both unilateral and bilateral benign interstitial cell tumors in the testes of males at the high-dose. There also was a dose-related increasing trend and a significant difference in the pairwise comparison of the high-dose group compared to control for testicular tumors, exceeding th e historical control incidence.

Carcinogenicity in Rodents (Mouse)

In a carcinogenicity study (MRID 42825002), iprodione (95.7% purity) was administered in the diet to 50 Crl: CD-1 (ICR) BR mice/sex/dose fo r 99 weeks at nominal doses of 0, 160, 800 and 400 ppm, which were equivalent to doses of 0, 23, 115 and 604 mg/kg/day for males and 0, 27, 138 and 793 mg/kg/day for females. An add itional 15 mice/sex/group were administered iprodione for 52 weeks (interim sacrifice).

The statistical evaluation of mortality indi cated no significant incr emental changes with increasing dose in either sex, although the high -dose group displayed the highest mortality rate for both sexes. Food consumption and clinical si gns were comparable among the groups for bothIPRODIONE FINAL

Page 55 of 66 sexes. Body weight gains were reduced (NS) in the high-dose group by 14% in males and 11% in
females.

No treatment-related changes were se en in absolute or relative pituitary weights at any dose. At Week 52, absolute thyroid weight was decreased (p<0.05) by 29-33% at all doses in females, and the relative thyroid weight wa s also decreased (NS) by 20–27%. In males, decreases (NS) in absolute ( ↓ 11%) and relative ( ↓ 9%) thyroid weights were seen onl y at the high dose. At terminal sacrifice, there were dose-dependent increases in thyroid weights in both se xes, but the increase only attained significance (p<0.05) in the high-dose males ( ↑ 18%). No histological changes were noted in the thyroid.

Absolute and relative adrenal weights were incr eased at the high dose at Week 52 in males ( ↑ 67– 78%, p<0.01) and females ( ↑ 14–17%, NS) and at Week 100 in males ( ↑ 14–21%, NS) and females ( ↑ 6–14%, NS). At terminal sacr ifice, adrenals were enlarg ed in high-dose males (4/50, 8%) and females (5/50, 10%) compared to cont rol males (1/50, 2%) and females (0%). In addition, the majority of high-dose females (12/13) displayed hypertrophy of the cells of the zona fasciculate; this lesion wa s not seen in any other dose group.

Liver weights (absolute, adjusted, and relative) were generally increased in a dose-dependent manner, but the increases only attained signifi cance (p<0.01) at 52 weeks for adjusted liver weights (male ↑ 52% and female ↑ 66%) and at 100 weeks for absolute liver weight in females ( ↑ 2.81-fold) and adjusted liver weights in males ( ↑ 2.88-fold). There was an increase (p<0.01) in
the incidence of liver tumors in both males (26/50) and females (21/50) at the high-dose compared to control (male, 7/50; females, 2/50), which was accompanied by increases in several liver lesions (centrilobular hepa tocyte enlargement/vacu olation, area(s) of enlarged eosinophilic hepatocytes, pigmented macropha ges, centrilobular ne crosis, and amyloid deposits). SGOT and SGPT concentrations were elevated at the high- dose in both sexes compared to control at the interim sacrifice (only time th ese enzymes were examined).

In males, no treatment-related changes were seen in absolute or relative weights of testes with epididymides at any dose. However, at terminal sacrifice, a greater number of masses were noted in the testes of the high-dose males (5/50; 10% ) compared to control males (1/50, 2%), and
smaller and/or flaccid testes were found more fr equently at the high dose (14/50; 28%) than in control (4/40; 8%). In addition, generalized vac uolation and hypertrophy of the interstitial cells
of the testes was seen at 52 w eeks at the high dose (9/12 versus 0/13 control) and at 100 weeks (p<0.01) at both the mid-dose (25/50; 50%) and hi gh-dose (27/50; 52%), versus control (4/50; 8%).

In females, uterine weights (absolute, adjuste d, and relative) were d ecreased in the high-dose group by 30–56% (NS) at 52 weeks and by 63–69% at 100 weeks, with significance beingIPRODIONE FINAL

Page 56 of 66 attained in the adjusted uterine weight (p<0.01, ↓ 69%). At terminal sacrifice, fewer high-dose females (16/50; 32%) displayed uteri that were thickened compared to control (39/50; 78%). Absolute and relative ovarian weights were decr eased at 52 weeks at all doses, but there was no dose response. At 100 weeks, absolute and re lative ovary weights were decreased by 53% (NS) at the high dose. At the high dose, luteini zation of the interstitial cells of the ovary was seen at 52 weeks (8/13, versus 0/13 in control) and at 100 weeks (23/50 ve rsus 6/50 in control). In addition, the incidence of luteoma of the ova ries was increased (p<0.05) at the high dose (5/50; 10%) compared to control (0/50; 0%).

The LOAEL is 800 ppm (male 115 mg/kg/day; fe males 138 mg/kg/day), based on the increased incidence of centrilobular hepatocyte enlargemen t in females and the increased incidence of generalized vacuolation/ hypertroph y of the interstitial cells in the testes of males. The NOAEL is 160 ppm (male 23 mg/kg/da y; female 27 mg/kg/day).

Chronic Oral Toxicity in Non-Rodents (Dog)

In a chronic feeding study (MRIDs 00144391 and MRID 41327001), 6 Beagle dogs/sex/group were administered iprodione ( 86.5% purity) via the diet at no minal doses of 0, 100 ppm [male 4.1/female 4.3 mg/kg/day], 600 ppm [male 24.9/female 28.3 mg /kg/day], or 3600 ppm [male 145.3/female 152.5 mg/kg/day] for 12 months. Ther e were no treatment-related deaths, and no
adverse effects were observed on bo dy weight, food consumption, or cl inical signs in either sex.

At the high-dose, there were incr eases in both sexes in absolute a nd relative liver weights [males, 41% and 48%; females, 18% (NS) and 28%], and in alkaline phosphatase, SGOT, SGPT and LDH enzymes. In addition, absolute and relativ e adrenal weights were increased (p<0.05 or p<0.01) in high-dose males (37% and 42%) and females (43% and 51%). Males also had decreases in both absolute (mid-dose, ↓ 22%, p<0.05; high-dose, ↓ 41%, p<0.01) and relative (high-dose, ↓ 39%, p<0.01) prostate weights. At the mid- and high-doses , males also displayed an increased number of erythr ocytes with Heinz bodies.

No treatment-related changes were seen in the absolute and relative weights of the testes, ovaries, uterus, thyroid or pituitary glands, and no treatment-related hi stopathological changes were seen in the testes, epididymides, prosta te, ovaries, uterus, cervix, mammary glands, and thyroid, adrenal, or pituitary glands.

The NOAEL in this study is 100 ppm (males 4.1/ females 4.3 mg/kg/day), and the LOAEL is 600 ppm (males, 24.9/females, 28.3 mg/kg/day), based on decreased prostate wei ght and an increased incidence of erythrocytes with Heinz bodies.IPRODIONE FINAL

Page 57 of 66 In a second complementary chronic feedi ng study (MRID 42211101), 6 Beagle dogs/sex/group were administered iprodione ( 96.1% purity) in the diet at nominal doses of 0, 200 ppm [males 7.8/females 9.1 mg/kg/day], 300 ppm [males 12 .4/females 13.1 mg/kg/day], 400 ppm [males 17.5/females18.4 mg/kg/day], and 600 ppm [males 24.6/females 26.4 mg/kg/day] for 12 months. There were no treatment-related deaths, and no a dverse effects were observed on clinical signs, body weight/gain, and food consumpti on in either sex. No treatmen t-related changes were seen
in the absolute and relative we ights of the prostate, adrenal an d kidneys; no other organs were weighed. No treatment-related hist opathological changes were seen in the prostate or adrenal glands; no other organs were examined.

At the high-dose, decreases were observed in the red blood cell parameters (hemoglobin, hematocrit, and red blood cells). The NOAE L for systemic toxicity is 400 ppm [males17.5/females 18.4 mg/kg/day], and the LOAEL is 600 ppm [males 24.6/females 26.4 mg/kg/day], based on decreased red blood cell values. When both chronic dog studies are considered together, the NOAEL is 400 ppm [~18 mg/kg/day].IPRODIONE FINAL

Page 58 of 66 Special Mechanistic Studies – Effe cts on Hormone Concentrations

In a special study (Chuzel, 1998) designed to measure the eff ect of iprodione on plasma
testosterone and lute inizing hormone (LH), iprodione ( 96.5% purity) was administered to 10 male SD Crl:CD [SD] BR rats/group via gavage in 0.4% aqueous methylcellulose as a single dose at 0, 70, and 300 mg/kg; a positive contro l group of 10 male rats was administered ketoconazole at 300 mg/kg. There were five repl icates of the negative control and iprodione groups, and one replicate was sampled at each of five time points (2, 4, 6, 10, and 24 hours post dose). There were two replicates of the positiv e control, and one replicate was sampled after either 4 or 24 hours post-dose.

There was an apparent dose-related decrease in testosterone concentr ation at both the low ( ↓ 59%) and high ( ↓ 75%) doses at 2 hours post-dose, a nd a significant increase in LH ( ↑ 53%, p<0.05) at the high dose. At 4 hours post-dose, there was also a dose-related increase in LH concentration at both the low ( ↑ 24%) and high ( ↑ 33%) doses, along with decreases in

testosterone concentr ation at the low ( ↓ 29%, NS) and high ( ↓ 59%, p<0.05) dose.

In another special study (Chuzel, 2001) examining iprodione effects on plasma concentrations of testosterone and LH, male SD ra ts (20/dose group/sampling interv al) were given a single oral dose of iprodione (97.4% purity) in 0.4% methylcellulose at 0, 70 and 300 mg/kg via gavage. Blood samples were taken at 0.5, 1, 2, 4 or 24 hour s post-dose and analyzed for testosterone and

LH.

Testosterone levels were decreased (p <0.01) at both doses by 0.5 hour post-dose ( ↓ 48 to 49%) with the measured peak effect occu rring 2 hours post-dose at the low dose ( ↓ 64%) and 1 hour post-dose at the high dose ( ↓ 67%). By 4 hours post-dose, test osterone concentrations were decreased by 35% (NS) and 54% at the low and high dose, re spectively, and testosterone
concentrations in treated males returned to control levels within 24 hours post-dose. Changes in LH concentrations lagged behind changes in testos terone concentrations, with increases in LH
concentrations observed at 2 and 4 hours post-dose at the low ( ↑ 15 and 10%, NS) and high ( ↑ 38 and 30%, p<0.01) dose.

In another related special st udy examining iprodione effects on plasma concentrations of
testosterone and LH after re peated dosing, (Chuzel, 2002b), 15 male SD rats/group/sampling time were administered iprodion e (97.4% purity) via gavage in 0.4% methylcellulose for 14 days at doses of 0, 6, 70 or 300 mg/kg/day. After 14 da ys of treatment, blood samples were taken at 1, 2, 4 and 24 hours after the last dose, and testoster one and LH concentrations were measured in plasma.IPRODIONE FINAL

Page 59 of 66 There were no treatment-related deaths. Decreased body weights ( ↓ 6 to 7%) were observed only at the high dose at study termina tion. Decreased testosterone concen trations were observed at the low dose by 1 hour ( ↓ 17%, NS) and 2 hours ( ↓ 9%, NS) post-dose; at the intermediate dose at 1 hour ( ↓ 26%, NS) and 2 hours ( ↓ 44%, p<0.05) post-dose; and at the high dose at 1 hour ( ↓ 36%, p<0.05), 2 hours ( ↓ 37%, p<0.05), and 4 hours ( ↓ 48%, p<0.01) post-dose. The time to attain “peak-effect” increased with the increase in dose; i.e. , 1 hour at the low dose, 2 hours at the intermediate dose, and 4 hours at the high dose.

At the low dose, increased LH concentrations ( ↑ 25%, NS) were observed at 1 hour post-dose. At the intermediate dose, LH concentrations were increased by 32% (NS) and 24% (p<0.01) at 2 and 4 hours post-dose, respectively. At the high dose, LH concentrations were increased by 21% (NS), 39% (NS), 54% (p<0.01) and 15% (NS) at 1, 2, 4 and 24 hours post-dose, respectively. The apparent peak-effect time for increased LH concentrations was 1 hour at the low dose, 2 hours at the intermediate dose, and 4 hours at the high dose. Thes e data indicate that repeated oral dosing with iprodione result s in decreases in plasma testosterone concentrations with a
concomitant increase in LH concentrations. Th e “peak” effects in hormone concentrations are attained rapidly after dosing, with the peak effect occurring at la ter intervals at higher doses.
Hormone concentrations returned to c ontrol levels within 24 hours post-dose.

Special Mechanistic Studies – Testicular Effects

In an in vivo study designed to examine the effect of iprodione on Leydi g cell proliferation (Kennel, 2002a), 15 male SD rats/group were ad ministered iprodione (97.4% purity) via gavage in 0.4% methylcellulose for 14 days at 0, 6, 70, or 300 mg/kg/day. A similar group of 15 male SD rats was administered flutamide, an androge n receptor antagonist, via gavage for 14 days at 150 mg/kg/day as a positive control. Seven days before the final sacrifice, animals were exposed to bromodeoxyuridine (Brd U) in sterile saline with osmotic mini pumps in order to examine Leydig cell prolifer ation (cycling) using imm unohistochemical staining.

Terminal body weights, brain weig hts (absolute and relative), and absolute testes weights were comparable among the iprodione and control gr oups. However, relative testes weight was increased (p<0.05) by 6% at the high dose, and flutamide decreased (p<0.01) absolute testes weight by 13%, compared to control.

There was a significant, dose-related increase in the mean BrdU labeling index at the intermediate ( ↑ 36%, p<0.05) and high ( ↑ 74%, p<0.01) doses during th e second-week assessment period. At the low dose, there was also a slight increase ( ↑ 14%, NS) that was considered treatment-related. The positive control flutamide displayed the expected increase in the Leydig cell proliferation index ( ↑ 3.4-fold). The findings dem onstrated that iprodione treatment can result in a dose-dependent increase in Leydig cell proliferation in the testis.IPRODIONE FINAL

Page 60 of 66 In an ex vivo study designed to examine the effect of iprodione exposure on basal testosterone
secretion from testicular s ections following a hCG Endoc rine Challenge Test (Fail et al , 1996a),
testicular sections were obtai ned from male SD rats (12/dos e group) administered iprodione (97.3% purity) in the diet for 14 days at dos es of 0 or 3000 ppm (~175 mg/kg/day). During the in

vivo phase of the study, rats were monitored for survival, clinical signs, body weight, and food consumption. At sacrifice the following organs /tissues were collected and weighed: liver, kidneys, adrenals, testes, epididymis, and total accessory sex organs (TASO). Blood samples were also collected for analysis of testoste rone and LH plasma concentrations. For the in vitro phase of the study, testis sections from each rat were incubated with 0, 1, 10, or 100 μ g/mL iprodione for one hour, and half the sections fr om each group were challenged with hCG and the
other half were monitored for basal testosterone secretion. Testosterone concentrations in the
culture media were monitored hour ly for 3 hours after challenge.

All rats survived during the in vivo phase and no clinical signs were noted in the iprodionetreated group. Body weights were decreased (p <0.05) by 10% compared to control in the treatment group. Absolute and relative liver wei ght was not affected by treatment, but absolute kidney weight was decreased (p<0.05) by 11%, a nd absolute and related adrenal weights were increased (p<0.05) by 27 and 40%, respectively. Re lative testis weights we re increased (p<0.05) by 11% and absolute epididymis weight was decr eased (p<0.05) by 7%. In addition, absolute and relative TASO weights were decreased (p<0.05) by 23 and 20%, respectively. At sacrifice, plasma concentrations of both testosterone and LH were increased by 32% (NS) and 110% (p<0.05), respectively. The signifi cant increase in plasma LH at necropsy suggests a possible
stimulation of the homeostatic mechanism.

In the in vitro phase of the study, there was a dose-relate d reduction in testosterone secretion from testicular sections incubated with ipro dione, with and without hCG stimulation. Prior in

vivo exposure of the rats to iprodione for 14 days app eared to have little ef fect on the secretion of testosterone, with and without hCG stimulation, aside from testosterone concentrations being slightly higher at t=0 in testes from in vivo -treated rats. Under the c onditions of this 14-day study, iprodione was shown to produce a reduction in testosterone secretion from testicular
sections after in vitro incubation with iprodion e. Prior exposure of male rats to iprodione in vivo via the diet for 14 days did not alter the reductio n in testosterone secr etion observed in their
testicular sections exposed to iprodione in vitro . Although the in vitro inhibition appeared to be dose-related, it appears that a maximum res ponse may have occurred between the 10 and 100

μ g/mL concentrations. Although the data provide som e evidence for the possible effect of iprodione on the testis, the pictur e of what is occurring in the tes tis of iprodione-treated rats is not complete. Although it appears th at the premise is that ipr odione produces testosterone biosynthesis inhibition, re sulting ultimately in the increased incidence of Leydig cell tumors,
there are inconsistencies in the in vitro and in vivo data.IPRODIONE FINAL

Page 61 of 66 As a complement to the above study, an in vivo study (Fail et al ., 1996b) was conducted to assess
the effects of in vivo iprodione exposure on plasma and testicular homogenate testosterone
concentrations in the male rat after an hCG Endocrine Challenge Test. For this study, male SD rats (15/dose group/sampling interv al) were administered iprodione in the diet at 0 ppm or 3000 ppm for 2, 7, or 14 consecutive days. The ipr odione dietary exposure was equivalent to 153.04 mg/kg/day by Day 2, 172.37 mg/kg/day by Da y 7, and 186.16 mg/kg/day by Day 14. For the endocrine challenge test, the rats from each group were administered hCG at 0.8 IU/100 g body weight intravenously via the ta il vein after ip rodione exposure, followed by sacrifice at 1 hour after hCG treatment. At sacrifice, the following orga ns/tissues were collected and weighed: liver, kidneys, adrenals, testes, epididymis, and tota l accessory sex organs (TASO). Blood samples were also collected, and plasma and testis homogenates were analyzed for testosterone
concentrations.

Body weights were decreased 5%, 10% and 13% at 2, 7 and 14 days, respectively, following treatment with 3000 ppm iprodion e. After 14 days of iprodi one treatment at 3000 ppm, the following changes (p<0.05) were noted in abso lute and/or relative or gan weights: liver ( ↓ 8% absolute, ↑ 6% relative), kidneys ( ↓ 9% absolute), adrenals ( ↑ 27% absolute, ↑ 46% relative), total testes ( ↑ 16% relative), total epididymides ( ↓ 9% absolute), TASO ( ↓ 27% absolute, ↓ 16% relative). There were no significant differences in testosterone concentrations at any sampling interval in either peri pheral plasma or testicular homogenate at one hour after the hCG challenge.
Under the conditions of this st udy, iprodione did not produce apparent alterations in testicular function after 14 days of dietary exposure at 3000 ppm (~175 mg/kg/day); however, the study
did not assess LH concentrations.

In an in vitro study (Benhamed, 1995) with cultured porci ne Leydig cells, ipr odione and two of its metabolites, RP36112 (99.2%) and RP36115 (96.7%), were shown to inhibit hCG-stimulated
testosterone secretion in a c oncentration range of 1 to 10 μ g/mL iprodione. Inhibition by iprodione was observed after short-term exposur e (3 hours), and the inhibitory effects were
similar to those observed with th e fungicide ketoconazole. The inhi bitory effects do not appear to be related to Leydig cell damage because the rem oval of iprodione from the culture medium for 72 hours resulted in the recovery of the cells abilit y to secrete testosterone after hCG stimulation.

In another in vitro study using immature porcine cultu red Leydig cells (Benhamed, 1996), iprodione (99.7%) and two of its m etabolites, RP36112 (99.2%) and RP36115 (96.7%), inhibited
testosterone secretion when Le ydig cells were stimulated with (1) hCG, (2) with drugs that enhance cAMP production [(a) cholera toxin, which stimulates the stimulatory guanine nucleotide-binding protein ( Gs ) ; (b) forskolin, which stimulates adenylate cyclase catalytic unit], and (3) with a cAMP analog (8-bromo-cAMP) . Because there were no effects observed on gonadotropin-stimulated cAMP production with ip rodione, it is hypothesized that the inhibitionIPRODIONE FINAL

Page 62 of 66 of testosterone secretion by ipr odione is downstream from cAMP production. At the next step in testosterone biosynthesis, inhi bition of testosterone secreti on by iprodione was not observed
when the substrate 22ROHCT was added to the cu lture medium, which indicates that the step
that is inhibited is locate d between the cAMP production a nd the movement/penetration of
cholesterol into the mitochondria. Because 22R OHCT is a cholesterol substrate that passes through the mitochondrial membrane without the need of an active transport system, the
sensitive site for inhibition of testosterone synthesis by iprodione (or RP 36115) may be the transport/availability of cholesterol substrate for the cholesterol side chain cleavage enzyme. The RP 36112 metabolite appears to act downs tream from the cholesterol step; i.e. , at the level of steroidogenic enzyme 17- α hydroxylase/17, 20 lyase. Iprodione a nd its metabolites appear to modulate Leydig cell steroidogenesis by interfering at the level of cholesterol transport and/or steroidogenic enzyme activity.

Avian Reproduction

In avian reproduction study (MRID 00086840), 6-month old Mallard ducks ( Anas platyrhynchos ) were dosed with iprodione (95% purity) in the feed at nominal concentrations of 100, 300 or 1000 ppm for up to 20 weeks. Each dose group consisted of 12 ducks/sex, with a single drake and hen being paired in 12 pe ns/dose group. The photoperi od was eight hours of light per day during the first 7 weeks (pre egg-la ying), and was increased to 17 hours of light per day for the remainder of the study. All adult bird s were observed daily for mortality, signs of toxicity, and abnormal behavior. Body weights were recorded at study initiation and at Weeks 2, 4, 6, 8, and at study termination, but not during egg-laying. Weekly, during egg-laying, one egg from every other pen was randomly selected for egg weight and eggshell thickness measurement. Feed consumption was measured every 2 weeks. Eggs were collected daily, marked, cleaned and
then stored at 56.0±1.5°F and 85% relative humid ity. Weekly, the eggs were placed in an incubator. Prior to incubation, a ll eggs were candled to detect cracks; on Day 14 of incubation, embryo viability was measured and E. coli -contaminated eggs were removed; on Day 21, embryo survival was determined; on Day 23, eggs were placed in a hatcher; on Day 28, all hatchlings, unhatched eggs, and eggshells were removed and the av erage body weight of representative hatchlings dete rmined. Hatchlings were housed in battery brooders, and weighed at 14-days old.

No adult mortalities occurred in any test group. At the intermediate dose, one hen exhibited loss
of coordination and lower limb weakness for 2 da ys and one hen had swollen tissues. At the high dose, two drakes and one hen exhibited lower limb weakness; one hen exhibited loss of coordination and lower limb weakness; one hen exhibited loss of c oordination, lower limb weakness and rigidity, and wing droop. No statistically significant e ffects were seen at any test level on adult body weight or average feed consumption. Mean eggshell thickness wasIPRODIONE FINAL

Page 63 of 66 approximately the same at all test concen tration (0.341 to 0.343 mm), and there was no difference in the number of eggs laid or ha tched between the treated and control group.

No significant overall impairment (p>0.05) was seen, across all reproductive parameters (from eggs laid to 14-day old survivors), at any dos e. Differences (p<0.05) were detected between treatment groups and control, bu t only at the low and intermed iate doses, where reproductive success was similar or increased compared to co ntrol for all individual parameters. At the high dose, although no overall significant reproductive di fference relative to control was detected (p>0.05), fewer hatchings survived to Day 14 (p<0.05). Based on the reduction in 14-day hatching survival, the LOAEC for reproductive effects is 1000 ppm, and the NOAEC is 300

ppm.

In another avian reproduction study (MRID 00099126), 6-month old Bobwhite Quails ( Colinus virginianus ) were dosed with iprodione (95% purity) in the feed at nominal concentrations of 100, 300 or 1000 ppm for up to 20 weeks. Each dose group consisted of 36 quails, with a single cock and two hens being paired in 12 pens/ dose group. The photoperiod was eight hours of light per day during the first 7 weeks (pre egg-laying) , and was increased to 17 hours of light per day for the remainder of the study. Adults r eceived water and the appropriate diet ad libitum . The adults were observed daily for mortality, signs of toxicity, and abnorma l behavior. Body weights were recorded at study initiation and at Weeks 2, 4, 6, 8, and at study termination, but not during egg-laying. Weekly, during egg-laying, one egg from every other pen was randomly selected for egg weight and eggshell thickness measurement. Feed consumption was measured every 2 weeks. Eggs were collected daily, marked, cleaned and then stored at 56.0±1.5°F and 85% relative humidity. The eggs were placed in an inc ubator weekly. All eggs we re candled to detect cracks prior to incubatio n and then measured for viability on Day 11 of incubation. On Day 21, embryo survival was determined and eggs were placed in a hatcher. On Day 25, all hatchlings, unhatched eggs, and eggshells were removed a nd the average body weight of representative hatchlings determined. Forty hatchlings from each week lot were randomly chosen for rearing. When the number of hatchlings in a given week lot was less than 40, all ha tchlings were reared. Hatchlings were housed in battery brooders until 14-days old and then weighed.

Adult mortalities occurred at all doses and were not consider ed to be treatment related;
behavioral effects were also seen in a ll treatment groups. Adult body weights and feed consumption in the treated groups were comp arable to the control group. There was no
treatment-related effect on the prevalence of cracked eggs, an d mean eggshell thickness in treatment groups was greater than or equal to control. No reproductive impairment (p>0.05) was seen at the low and high doses. The study author s reported a significant (p<0.05) decrease in the number of eggs laid at the high dose (524 vs. 692 in control), a reduction in hatchling body weight (6.0 g. vs. 6.3 g. in control), and a significant (p< 0.01) decrease in per cent of hatchlings of eggs set. However, re-analysis of the data confirmed only a significa nt (p<0.05) reduction inIPRODIONE FINAL

Page 64 of 66 hatchling body weight. The reproductive NOAEC for this study was 300 ppm, and the LOAEC was 1000 ppm.

Fish Early Life Stage (ELS) Study

In a fish ELS study (MRID 40550801), embryonic/larval fathead minnows ( P. promelas ), <24- hours old (2 × 60 embryos/dose group), were expo sed for 34 days in glass aquaria under flowthrough conditions to iprodione (100% purity) at nominal concentrations of 2.5, 1.2, 0.62 0.31, or 0.15 mg/L; these concentrations were equivalent to mean-meas ured concentrations of 2.2, 1.1, 0.55, 0.26 and 0.13 mg/L, respectively. Th e test system was maintained at 25±1°C at a pH of 7.2 to 7.3. Embryos were evaluated for time to ha tching and survival until hatching was complete, and larvae (Day 4) were evalua ted for survival, behavior and appearance twice weekly. At 30 days post-hatch, the percentage survival was dete rmined, and larvae were individually measured for wet body weight and length.

As there were no differences between the clean wa ter and solvent controls, data from the control groups was pooled for comparison with the treate d groups. Survival at the end of the hatching period was reduced by 11% (p<0.05) at the highest tested concentration (2 .2 mg/L). After the 30- day post-hatch exposure, larval survival was decreased (p<0.05) by 96% , 42% and 10% in the 2.2, 1.1 and 0.55 mg/L groups, respectively, compared to the pooled control. For the surviving larvae, wet body weights were reduced (p <0.05) by 70% and 26% at 2.2 and 1.1 mg/L, respectively, and body length was reduced (p<0.05) by 31% and 11%, respectively. Based on reductions in larvae survival, the LOAEC for iprodione was 0.55 mg/L and the NOAEC was 0.26 mg/L.IPRODIONE FINAL

Page 65 of 66 APPENDIX 3: References Not Utilized in the Iprodione WoE Analysis

In 2009, after public review and comment, a fina l list of 67 chemicals and schedule for issuing
Test Orders for the EDSP Tier 1 screening battery was made av ailable in a Federal Register Notice issued October 21, 2009 (74 FR 54422). The ag ency’s review of the initial data submitted as “other scientifically releva nt information (OSRI) was provided in the Report of the Endocrine Disruptor Review Team (USEPA, 2010). Beginning in 2011, the agency has reviewed data cited as “OSRI which included Part 158 studies previously submitted to the agency for registra tion/reregistration, published literature articles and/or Tier 1 assays. The agency also conduc ted a more recent s earch (2009 to 2014) of
available scientific literature fo r any additional relevant informa tion for their weight of evidence (WoE) evaluations. These articles were evalua ted in accordance with the agencies Evaluation Guidelines for Ecological Toxicity Data in Open Literature, May 2011 ( http://www.epa.gov/pesticides/s cience/efed/policy_guidance/team _authors/endangered_species _reregistration_workgroup/PDF_ro t/esa_evaluation_open_literature.pdf ) and the 2012 Guidance for considering and Using Open Literature To xicity Studies to Suppor t Human Health Risk Assessment ( http://www.epa.gov/pesticides /science/lit-studies.pdf).

The following published and unpublished references were considered for use in the WoE analysis for Iprodione but were not utilized due to one or more of the following reasons: 1) the article was not available in English; 2) the com pound of interest was not us ed in the study; 3) the test material was not adequately described; 4) a formulated end-use product or mixture of chemicals was utilized as the test material; 5) only acute mortality toxicity data were provided; 6) the experimental conditions were not adequate ly described; 7) only an abstract of the study was available; 8) the reference is a review article or book chapte r and does not contain primary study data; 9) insufficient information was availabl e to adequately assess th e validity of the study results; 10) the 40 CFR Part 158 guideline study wa s classified as unacceptable/inadequate; 11) the study dealt only with non- EDSP assay development; 12) no specific endocrine-related endpoints were assessed in the study; and 13) th e study contained only data on invertebrates.

Benhamed, M. (1996) Effects of iprodione and its metabolites RP 36112 and RP 36115 on
testosterone secretion in cultu red Leydig cells: Sites of action. INSERM U 407, Sophia Antipolis Cedex, France. INSERMIU 407/96001, C024294. October 21, 1996. (MRID 44171901)

de Groene, E.M. (1999) Human androgen recepto r binding assay with th e fungicide iprodione and 7 mammalian metabolites. TNO Voeding, TNO Nutrition and Food Research Institute, Analytical Sciences Division, Zeist, The Netherlands. TNO report V99.616, Project no. 50436. June 23, 1999.IPRODIONE FINAL

Page 66 of 66 Fisher, P. (1998) Quantification of iprodione and metabolites in th e plasma and testes of the rat following a single oral administration of [ 14 C]-iprodione. Rhone-Poulenc Agro CRSA, Cedex, France. Study no. SA98326, B003653, November 30, 1998. (MRID 44750701)