exposure risks for mixer/loaders are not of concern (i.e., MOEs>100),
provided the mixer/loaders wear protective gloves as directed on the
label.  Because the endpoint used in this assessment is from an oral
study, the estimated dermal and inhalation exposures were adjusted by
applying a 100 percent dermal and inhalation absorption rate,
respectively.  As a result, the estimated risks (MOEs ) are considered
highly conservative.

HED has no data to assess exposures to pilots using open cockpits.  The
only data available are for exposure to pilots in enclosed cockpits.
Therefore, risks to pilots are assessed using the engineering control
(enclosed cockpits) and baseline attire (long-sleeve shirt, long pants,
shoes, and socks); pilots are not required to wear protective gloves.
With this level of protection, there are no risks of concern for
applicators.

Table 9.1.1  Cyprosulfamide Occupational Short and Intermediate-Term
Dermal and Inhalation Exposures and Risks

Crop or Target	App Rate (lb ai/acre)a	Area Treated Daily (acres)b	Dermal
and Inhalation Unit Exposures

(mg/lb ai)	Doses (mg/kg/day)g	MOEsh	Combined MOEsi

Mixing/Loading Liquid Concentrates for Aerial Applications

corn 	0.141	1200	Dermal

Baselinec: 2.9

SL w/glovesd: 0.023	Dermal

Baseline: 7.00

SL w/gloves: 0.056	Dermal

Baseline: 8

SL w/gloves: 1,000	Baseline Dermal and Inhalation:

PPE – SL w/gloves + Baseline Inhalation:  1,000

	Inhalation

Baselinee: 0.0012	Inhalation

Baseline: 0.003 	Inhalation

Baseline:  19,000

	Mixing/Loading Liquid Concentrates for Grounboom  Applications

corn	0.141	200	Dermal

Baselinec: 2.9

SL w/glovesd: 0.023	Dermal

Baseline: 1.17

SL w/gloves: 0.0093	Dermal

Baseline: 48

SL w/gloves: 6,045

	Baseline Dermal and Inhalation:

PPE – SL w/gloves + Baseline Inhalation:  5,750

	Inhalation

Baselinee: 0.0012	Inhalation

Baseline: 0.00048 	Inhalation

Baseline: 116,000

	Applying Sprays via Aerial Equipment

corn 	0.141	1200	Dermal

Engineering controlf: 0.005	Dermal

Engineering control: 0.0121	Dermal

Engineering control:4,633 	Engineering control Dermal + Inhalation:
4,570

	Inhalation

Engineering control: 0.000068	Inhalation

Engineering control: 0.000164 	Inhalation

Engineering control: 340,700

	Flagging to Support Aerial Applications

corn 	0.141	350	Dermal

Baselinef: 0.01	Dermal

Baseline: 0.0078	Dermal

Baseline: 7500	Baseline Dermal and Inhalation: 7,250

	Inhalation

Baseline: 0.00035	Inhalation

Baseline: 0.00025	Inhalation

Baseline: 235,000

	Applying Sprays via Groundboom  Equipment

corn	0.141	200	Dermal

0.014	Dermal

0.00564	Dermal

9,929	 Dermal + Inhalation: 9,430

	Inhalation

0.00074	Inhalation:

0.000298	Inhalation

187,847

	Mixing/Loading Liquid and Applying with Handgun Sprayer

Turf 	0.027

Ib ai/A	5A	0.50 (gloves)	Dermal:

0.00096	Dermal

60,500	48,000

	0.0019	Inhalation:

0.0002565	Inhalation

226,000

	Mixing/Loading Liquid and Applying with Low-Pressure Handwand

Turf 	0.01

lb ai/gal	40

gals	100	Dermal:

0.571	Dermal

102	100

	0.03	Inhalation:

0.00017	Inhalation

341,000

	Mixing/Loading Liquid and Applying with High-Pressure Handwand

Ornamentals	0.01

lb ai/gal	1000

gals	2.5 (gloves)	Dermal:

0.357	Dermal

163	155

	0.12	Inhalation:

0.0171	Inhalation

3,400

	 Mixing/Loading Liquid and Applying with Backpack Sprayer

Turf and Ornamentals	0.01

lb ai/gal	40

gals	2.5 (gloves)	Dermal:

0.0142	Dermal

4100	4,036

	0.03	Inhalation:

0.000171	Inhalation

338,000

	 Mixing/Loading water soluble granular and Applying with Backpack
Sprayer, handgun sprayer, low pressure handwand or high pressure
handwand

Turf 	0.027	5A or 40 gals	No Data	Dermal:

Not Assessed	Dermal

Not Assessed	Not Assessed

	No Data	Inhalation:

Not Assessed	Inhalation

Not Assessed

	a	Application rates are the maximum application rates determined from
proposed labels for cyprosulfamide

b	Amount handled per day values are HED estimates of acres treated per
day based on Exposure SAC SOP #9 “Standard Values for Daily Acres
Treated in Agriculture,” industry sources, and HED estimates.

c	Baseline Dermal:  Long-sleeve shirt, long pants, and no gloves.

d	Dermal SL w/gloves: Single layer plus chemical-resistant gloves.

e	Baseline Inhalation: no respirator.

f	Engineering control: enclosed cockpit

g	Dose (mg/kg/day) = Unit exposure(mg/lb ai) x App Rate (lb ai/acre) x
Area Treated (acres/day) x  %Absorption (100% dermal and 100%
inhalation) / Body weight (70 kg).

h	MOE = LOAEL/Dose; where the short- and intermediate-term dermal and
inhalation NOAEL = 58 mg/kg/day

i	Combined MOEs =LOAEL / (Dermal dose + Inhalation Dose)

Seed Treatment

There is potential for occupational exposure from the proposed use
(i.e., Sorghum) as a seed treatment.  It is anticipated that the
following scenarios could result in handler exposure. The exposure
assessment scenarios are as follows:

Loading (open)/applying a liquid formulation for seed treatment;

Sewing bags of treated seed;

Bagging treated seeds;

Multiple activities involved in seed treatment; and

Planting of treated seed.

The unit exposures are based on SOPs #14 and #15.  SOP #14 contains all
the known scenarios associated with seed treatment facilities and
planting treated seed.  The data contained in this document are for
worker exposure estimation and are mostly generated by the industry.
Exposure estimates are from actual seed treatment studies and are based
on physical factors or a handler scenario (e.g., commercial seed
treatment, on farm seed treatment, planters, etc).  The unit exposures
are based on a central tendency from multiple studies summarized in SOP
#14.

For pesticide handlers, it is HED standard practice to present estimates
of dermal exposure for “baseline” (i.e., workers wearing a single
layer of work clothing consisting of a long sleeved shirt, long pants,
shoes plus socks and no protective gloves), as well as for
“baseline” and the use of protective gloves or other Personal
Protective Equipment (PPE) as might be necessary.  The product label
directs workers involved with treated the seed to wear long-sleeve shirt
and long pants, chemical resistant footwear plus socks, chemical
resistant gloves, a chemical resistant apron, and a dust/mist filtering
respirator or a NIOSH approved respirator with an R, P or HE filter.
Baggers and bag sewers must wear a long-sleeved shirt and long pants,
shoes plus socks, chemical resistant gloves, and a dust/mist filtering
respirator or a NIOSH approved respirator with an R, P or HE filter.

HED believes most exposure durations will be short-term (1 - 30 days).
However, HED’s Science Advisory Council for Exposure (ExpoSAC)
maintains it is possible for commercial applicators to be exposed to
intermediate-term exposure durations (1 - 6 months).   In addition, the
short- and intermediate-term toxicological endpoints are the same,
therefore, the estimates of risk for short-term duration exposures are
protective of those for intermediate-term duration exposures.  Long-term
exposures are not expected, therefore, a long-term assessment was not
conducted.

The following equations were used to calculate seed treatment handler
exposure and risk:

(1) Inhalation Dose (mg/kg/day) = Rate (lb ai/lbs seed) x UE (mg/lb ai)
x AT (lbs/day) / BW (kg)

Where:

Rate (Application Rate)	=	Maximum application rate on product label (lb
ai/lbs seed);

UE (Unit Exposure)	=	Exposure value derived from SOP 15 (mg/lb ai);

	AT (Amount treated)	=	Max. lbs. of seed treated/day (lbs/day); and

            	BW			=	Body weight (kg).

(2) Inhalation MOE	=	NOAEL (mg/kg/day) / Inhalation Dose (mg/kg/day)

Table 9.1.2 presents the estimated risks for workers based on the short
and intermediate-term inhalation exposures at baseline. HED has
determined that risks are not of concern (i.e., MOEs>100).

Table 9.1.2  Occupational Handler Exposures and Risks

Inhalation Unit Exposures

(mg/lb ai)a	Application rate (lb ai/100 lb seed)	lbs seed treated or
planted/dayb	Short- and Intermediate-term  Doses (mg/kg/day)c	Short- and
Intermediate-term Dermal and Inhalation MOEsd	Total MOE

Dermal Unit Exposures (mg/lb ai)a

	Loading (open)/applying liquid for seed treatment

0.00034	0.052	718,000	Inhalation: 0.0018	Inhalation: 31000	450

0.0230

	Dermal:0.123	Dermal:460

	Sewing bags of treated seed

0.00023	0.052	718,000	Inhalation:0.00122	Inhalation: 46000	1633

0.0062

	Dermal:0.033	Dermal:1700

	Bagging seeds after seed treatment

0.00016	0.052	718,000	Inhalation:0.00085	Inhalation: 66000	1134

0.0091

	Dermal:0.049	Dermal:1200

	Multiple activities involved in seed treatment

0.0016	0.052	718,000	Inhalation:0.0085	Inhalation: 6600	240

0.042

Dermal:0.224	Dermal:250

	Planting of treated seed

0.0034	0.052	12,000	Inhalation:0.0003	Inhalation:185000	2500

0.2500

	Dermal:0.0223	Dermal:2500

	a	Baseline Inhalation and dermal unit exposures are taken from the
ExpoSAC Policy 14.

b	Lbs of seed treated or planted/day values are based on ExpoSAC Policy
15.

c	Inhalation and dermal dose (mg/kg/day) = [unit exposure (mg/lb ai) *
Appl. rate (lb ai/lbs seed) * Amount treated / body weight (70 kg)].

d	MOE = NOAEL (58 mg/kg/day) / inhalation or Dermal dose (mg/kg/day).
UF = 100.

9.2	Short- and Intermediate-term Postapplication Risk  TC \l2 "9.2
Short-and Intermediate-Term Postapplication Risk

There is the possibility for agricultural workers to have
postapplication exposure to cyprosulfamide following its proposed
agricultural crop uses.  Therefore, occupational postapplication risks
were assessed.

HED uses a concept known as the transfer coefficient to numerically
represent the postapplication exposures one would receive (generally
presented as cm2/hour).  A transfer coefficient is a measure of the
residue transferred from a treated surface to a person who is doing a
task or activity in a treated area.  These values are the ratio of an
exposure for a given task or activity to the amount of pesticide residue
on treated surfaces available for transfer.  HED has developed a series
of standard transfer coefficients that are unique for variety of job
tasks or activities that are used in lieu of chemical- and
scenario-specific data.  HED used Policy 003.1 Science Advisory Council
for Exposure Policy Regarding Agricultural Transfer Coefficients to
estimate transfer coefficients for the various tasks that
postapplication workers might perform on each crop grouping.  HED
assumes an 8 hour workday for postapplication workers and assumes 70
kilograms for an adult (average adult weight).

For cyprosulfamide, the exposure durations for non-cancer
postapplication risk assessment were short-term (30 days) and
intermediate-term (greater than 30 days up to several months).  However,
since the dermal toxicological endpoint of concern is the same for
short- and intermediate-term exposures, the short- and intermediate-term
postapplication risks are numerically identical.  HED has established
levels of concern (LOCs) for occupational postapplication risks –
margins of exposure of less than 100 for occupational non-cancer dermal
risks are a concern.

Inhalation exposures are thought to be negligible in outdoor
postapplication scenarios, since cyprosulfamide has low vapor pressure
and the dilution factor outdoors is considered infinite.

Occupational Postapplication Exposure

Since no postapplication data were submitted in support of this
registration action, exposures during postapplication activities were
estimated using dermal transfer coefficients from the Science Advisory
Council for Exposure Policy Number 3.1: Agricultural Transfer
Coefficients, August 2000, summarized in Table 9.2.1 below and the
following assumptions:

Assumptions:

Application Rate	= 	0.141lb ai/A

Exposure Duration	=	8 hours per day

Body Weight		=	70 kg

Dermal Absorption	= 	100%

Fraction of a.i. retained on foliage is assumed to be 20% (0.20) on day
zero (= % dislodgeable foliar residue, DFR, after initial treatment).
This fraction is assumed to further dissipate at the rate of 10% (0.10)
per day on following days.  These are default values established by
HED’s Science Advisory Council (SAC) for Exposure.

Table 9.2.1  Anticipated Postapplication Activities and Dermal Transfer
Coefficients

Proposed Crops	Policy Crop Group Category	Transfer Coefficients (cm2/hr)
Activities

Corn	Field / row crop, low / medium	100 	Hand weeding, scouting

1500 	Scouting, irrigation

Corn	Field / row crop, tall	400 	Scouting

1000	Irrigation

17,000 	Hand harvesting

Daily dermal exposures were calculated on each postapplication day after
application using the following equation:

DE(t) (mg/day) = (TR(t) (µg/cm2) x TC (cm2/hr) x Hr/Day)/1000 (µg/mg)

Where:

DE(t)	=	Daily exposure or amount deposited on the surface of the skin at
time (t) attributable for activity in a previously treated area, also
referred to as potential dose (mg ai/day);

	TR(t)	=	Transferable residues that can be dislodgeable foliar residue
at time “t” (µg/cm2);

	TC	=	Transfer Coefficient (cm2/hour); and

	Hr/day	=	Exposure duration meant to represent a typical workday
(hours).

Note that the (TR(t)) input may represent levels on the day of
application in the case of short-term risk calculations.  Once daily
exposures are calculated, the calculation of daily absorbed dose and the
resulting Margin of Exposures use the same algorithms that are described
above for the handler exposures.  These calculations are completed for
each day or appropriate block of time after application.  For
pre-emergence spray, the application rate of 0.141 lb ai/A is higher
than the recommended application rate of 0.013 lb ai/A for post
emergence application.  Therefore the risk presented is highly
conservative.

For corn, wheat and soybean applications, risks are not of concern
(i.e., MOE>100) on day 0 (REI = 12 hours) for all of the exposure
activities.  Table 9.2.2 presents a summary of occupational
postapplication risks associated with use of cyprosulfamide.

Table 9.2.2  Summary of Occupational Postapplication Risks for
Cyprosulfamide

Crop Grouping	Application rate

(lb ai/acre)	Transfer Coefficient (µg/cm2)	Day after Application	MOE at
Day 0

(Level of Concern = 100)

Corn	0.141	100 (Hand weeding, scouting)	0 (12 hours)	16,000

1500 (Scouting, irrigation)

1070

400 (Scouting)

4,000

1000 (Irrigation)

1,605

17,000 (Hand harvesting, Detasseling)

100

Cyprosulfamide is classified in acute toxicity category III for acute
dermal toxicity and category IV for primary eye irritation and primary
skin irritation.  It is not a dermal sensitizer.  A restricted entry
interval (REI) of 12 hours is appropriate and meets the requirements of
the Worker Protection Standard for Agricultural Pesticides (WPS).

10.0	Data Needs and Label Requirements  TC \l1 "10.0	Data Needs and
Label Requirements

10.1	Toxicology  TC \l2 "10.1	Toxicology

Guideline Number: 870.7800

Study Title:  Immunotoxicity

Rationale for Requiring the Data

This is a new data requirement under 40 CFR Part 158 as a part of the
data requirements for registration of a pesticide (food and non-food
uses).

The Immunotoxicity Test Guideline (OPPTS 870.7800) prescribes functional
immunotoxicity testing and is designed to evaluate the potential of a
repeated chemical exposure to produce adverse effects (i.e.,
suppression) on the immune system. Immunosuppression is a deficit in the
ability of the immune system to respond to a challenge of bacterial or
viral infections such as tuberculosis (TB), Severe Acquired Respiratory
Syndrome (SARS), or neoplasia.  Because the immune system is highly
complex, studies not specifically conducted to assess immunotoxic
endpoints are inadequate to characterize a pesticide’s potential
immunotoxicity.  While data from hematology, lymphoid organ weights, and
histopathology in routine chronic or subchronic toxicity studies may
offer useful information on potential immunotoxic effects, these
endpoints alone are insufficient to predict immunotoxicity.

Practical Utility of the Data

How will the data be used?

Immunotoxicity studies provide critical scientific information needed to
characterize potential hazard to the human population on the immune
system from pesticide exposure. Since epidemiologic data on the effects
of chemical exposures on immune parameters are limited and are
inadequate to characterize a pesticide’s potential immunotoxicity in
humans, animal studies are used as the most sensitive endpoint for risk
assessment.  These animal studies can be used to select endpoints and
doses for use in risk assessment of all exposure scenarios and are
considered a primary data source for reliable reference dose
calculation. For example, animal studies have demonstrated that
immunotoxicity in rodents is one of the more sensitive manifestations of
TCDD (2,3,7,8-tetrachlorodibenzo-p-dioxin) among developmental,
reproductive, and endocrinologic toxicities.  Additionally, the EPA has
established an oral reference dose (RfD) for tributyltin oxide (TBTO)
based on observed immunotoxicity in animal studies (IRIS, 1997).

How could the data impact the Agency's future decision-making?

If the immunotoxicity study shows that the test material poses either a
greater or a diminished risk than that given in the interim decision’s
conclusion, the risk assessments for the test material may need to be
revised to reflect the magnitude of potential risk derived from the new
data.

If the Agency does not have this data, a 10X database uncertainty factor
may be applied for conducting a risk assessment from the available
studies.

10.2	Residue Chemistry  TC \l2 "10.2	Residue Chemistry

860.1200 Directions for Use

The proposed PHI of 45 days for grain and stover is not supported by the
field trial data, which reflect longer PHIs than 45 days.  The
petitioner should modify the labels to clarify that the 45-day PHI
pertains only to corn forage (field and sweet) and sweet corn ears.
Because the product labels specify that applications may not be made
after the V12 growth stage of corn, a PHI is not required for corn RACs.

The label for the SC 480 Herbicide should be modified to clarify a
contradiction in the “Preplant Surface-Applied” application
directions and the “Restrictions and Precautions for Use” section.
The Restrictions section specifies that only one application may be made
to corn per season.  The instructions for Preplant surface-applied
application specify that the application may be split, with 60% of the
recommended rate applied prior to planting and the remaining 40% applied
at planting.  If the petitioner wishes to allow split
preplant/at-planting applications, the Restrictions section should be
modified accordingly.

The labels for the SC 465 and SC 480 Herbicides include instructions for
“Post-Harvest Non-crop” application but do not include any
application rates for this type of application.  The labels should be
modified to either propose application rates or to remove the
instructions for this type of application.

860.1340 Residue Analytical Method

A copy of the complete, detailed livestock method (UB-006-A06-01) with
corresponding registrant validation data should be submitted as an
independent method document.  Currently, only a synopsis of the method
is available (MRID # 47069738).

860.1380 Storage Stability

The final report of the storage stability studies, which are to include
storage intervals up to 18 months, must be submitted.

860.1550 Proposed Tolerances

The petitioner is required to submit a revised Section F to reflect the
revised tolerances and commodity definitions recommended in the
Introduction of this document.

860.1650 Submittal of Analytical Reference Standards

Analytical reference standards of cyprosulfamide metabolite AE 0852999
(M02) and isotopically-labeled standards of both parent and M02 to be
used as internal standards must be supplied to the EPA National
Pesticide Standards Repository.

10.3	Occupational and Residential Exposure  TC \l2 "10.3	Occupational
and Residential Exposure

None

References:  TC \l1 "References:

CARC Memo:  TXR 0054810, Jessica Kidwell, 2/29/2008

Residue Chemistry Summary Memo:  D341999, William Donovan, 5/28/08

Dietary Exposure Memo:  D351493, Seyed Tadayon, 5/6/08

ROCKS Memo:  D351495, George Kramer, 4/18/08

ToxSAC Memo: Jessica Kidwell, 4/24/2008

Occupational & Residential Exposure Memo:  D351494, Seyed Tadayon,
5/28/08

Appendix A:  Toxicology Assessment  TC \l1 "Appendix A:  Toxicology
Assessment

A.1	Toxicology Data Requirements TC \l2 "A.1  Toxicology Data
Requirements

The requirements (40 CFR 158.340) for food uses for cyprosulfamide are
in Table 1. Use of the new guideline numbers does not imply that the new
(1998) guideline protocols were used.

Test

	Technical

	Required	Satisfied

870.1100    Acute Oral Toxicity

870.1200    Acute Dermal Toxicity

870.1300    Acute Inhalation Toxicity

870.2400    Primary Eye Irritation

870.2500    Primary Dermal Irritation

870.2600    Dermal Sensitization		yes

yes

yes

yes

yes

yes	yes

yes

yes

yes

yes

yes

870.3100    Oral Subchronic (rodent)

870.3150    Oral Subchronic (nonrodent)

870.3200    21-Day Dermal

870.3250    90-Day Dermal

870.3465    90-Day Inhalation		Yes

Yes

No

No

No	Yes

Yes

N/A

N/A

N/A

870.3700a  Developmental Toxicity (rodent)

870.3700b  Developmental Toxicity (nonrodent)

870.3800    Reproduction		Yes

Yes

Yes	Yes

Yes

Yes

870.4100a  Chronic Toxicity (rodent)

870.4100b  Chronic Toxicity (nonrodent)

870.4200a  Oncogenicity (rat)

870.4200b  Oncogenicity (mouse)

870.4300    Chronic/Oncogenicity - rat	Yes

Yes

Yes

Yes

Yes	Yes

Yes

Yes

Yes

Yes

870.5100    Mutagenicity—Gene Mutation - bacterial

870.5300    Mutagenicity—Gene Mutation - mammalian

870.5xxx    Mutagenicity—Structural Chromosomal Aberrations

870.5xxx    Mutagenicity—Other Genotoxic Effects		Yes

Yes

Yes

Yes	Yes

Yes

Yes

Yes

870.6100a  Acute Delayed Neurotox. (hen)

870.6100b  90-Day Neurotoxicity (hen)

870.6200a  Acute Neurotox. Screening Battery (rat)

870.6200b  90-Day Neuro. Screening Battery (rat)

870.6300    Develop. Neuro		no

no

Yes

Yes

No	-

-

Yes

Yes

N/A

870.7485    General Metabolism

870.7600    Dermal Penetration		Yes

No	Yes

N/A

Special Studies for Ocular Effects

Acute Oral (rat)

Subchronic Oral (rat)

Six-month Oral (dog)		No	No

A.2  Toxicity Profiles

Table A.2.1	Acute Toxicity Profile - Test Substance

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

870.1100	Acute oral - rat	47069801

(2005)	LD50 > 2000 mg/kg	III

870.1200	Acute dermal - rat	47069802

(2005)	LD50 = 2000 mg/kg	III

870.1300	Acute inhalation - rat	47069803

(2004)	LC50 > 3513 mg/m3 	III

870.2400	Acute eye irritation - rabbit	47069805

(2004)	Not irritating	IV

870.2500	Acute dermal irritation - rabbit	47069804

(2005)	Not irritating	IV

870.2600	Skin sensitization - guinea pig 	47069806

(2005)	Not a sensitizer	N/A

Table A.2.2	Subchronic, Chronic and Other Toxicity Profile

Guideline No. 	Study Type	MRID No. (year)/ Classification /Doses	Results

870.3100

	90-Day oral toxicity - rat	47069808 (2003)

Acceptable/Guideline

0, 1000, 4000 or 12000 ppm

M: 0, 58, 240 or 720 mg/kg/day.

F: 0, 70, 281 or 859 mg/kg/day. 	NOAEL = 57.9 for males and 70.1 for
females mg/kg/day

LOAEL = 240 in males and 281 in females mg/kg/day based on and urinary
crystals/ hyperplasia in urinary system.

870.3100	90-day oral toxicity - mouse	47069809 (2003)

Acceptable/Guideline

0, 500, 2000 or 7000 ppm

M: 0, 79, 321 or 1110 mg/kg/day.

F: 0, 95, 398 or 1297 mg/kg/day	NOAEL = 398 mg/kg/day in females and
1110 mg/kg/day in males.

LOAEL = severity of lymphocytolysis in the thymus in females.

870.3150

	90-Day oral toxicity  - dog	47069811 (2005)

Acceptable/Guideline

0, 2500, 7500 or 15000 ppm

M: 0, 75, 221 or 416 mg/kg/day.

F: 0, 79, 221 or 341 mg/kg/day.	NOAEL = 221 mg/kg/day for both sexes

LOAEL = 416 mg/kg/day for males and 341 mg/kg/day for females. Based on
urinary tract effects.

870.3200

	21/28-Day dermal toxicity (species)	No study and not required.	N/A

870.3250

	90-Day dermal toxicity (species)	No study and not required. 	N/A

870.3465

	90-Day inhalation toxicity (species)	No Study and not required.	N/A

870.3700a

	Prenatal developmental in  rats	47069814 (2005)

Acceptable/Guideline

0, 60, 250 or 1000 mg/kg/day	Maternal NOAEL = 250 mg/kg/day

LOAEL = 1000 mg/kg/day based mainly on body weight effect.

Developmental NOAEL = 1000 mg/kg/day. No effects at the highest dose
tested.

870.3700b	Prenatal developmental in rabbits	47069812 (2005)

Acceptable/Guideline

0, 50, 125 or 500 mg/kg/day.	Maternal NOAEL = 125 mg/kg/day.

LOAEL = 500 mg/kg/day exceeds MTD based on deaths, body weight decrease,
reduced food consumption and kidney effects.

870.3700b

	Prenatal developmental in  rabbits	47069813 (2005)

Acceptable/Non-Guideline

0 or 250 mg/kg/day.	Maternal NOAEL =  Not established.

LOAEL = 250 mg/kg/day based on  reduced body weight and food consumption
and kidney effects.

Developmental LOAEL > 250 mg/kg/day.  mg/kg/day.  No effects at highest
dose tested.

870.3800

	Reproduction and fertility effects

In rats	47069815 (2007)

Acceptable/Guideline

0, 480, 2400 or 12000 ppm.

M: 0, 39.2, 202.3 or 1006.9 for FO and 0, 50.3, 251.5 or 1292.4 for F1
mg/kg/day.

F: 0, 55.3, 260.3 or 1350.2 for FO and 0, 68.5, 323.7 or 1696.5 for F1
mg/kg/day. 	Parental/Systemic NOAEL = 39 for males and 55 for females
mg/kg/day

LOAEL = 202 for males and 260 for females  mg/kg/day based on organ
weight changes in spleen and urinary tract. .

Reproductive NOAEL = 202 for males and 260 for females mg/kg/day

LOAEL = 1006.9 for males and 1350.2 for females mg/kg/day based on
reduced rearing index.

Offspring NOAEL = 202 for males and 260 for females mg/kg/day

LOAEL = 1006.9 for males and 1350.2 for females mg/kg/day based on pup
body weight decrease.

870.4100a	Chronic toxicity in the rat. 	See 870.4300.

870.4100b

	Chronic toxicity (dog)	47069816 (2006)

Acceptable/Guideline

0, 1000, 2500 or 8000 ppm.

M: 0, 29, 66 or 226 mg/kg/day

F: 0, 28, 67 or 242 mg/kg/day. 	NOAEL = 66 in males and 67 in females
mg/kg/day

LOAEL = 226 in males and 242 in females mg/kg/day based on urothelial
effects.

870.4200

	Carcinogenicity

(mouse)	47069818 (2006)

Acceptable/Guideline

0, 350, 2000 or 3500 ppm.

M: 0, 50, 287 or 506 mg/kg/day.

F: 0, 63, 354 or 616 mg/kg/day. 	NOAEL = 50 in males and 354 in females
mg/kg/day

LOAEL = 287 in males and 616 in females mg/kg/day based on urothelial
effects.

Transitional cell papilloma in female urinary bladder.  Histiocytic
sarcoma in females.

870.4300. 	Combined chronic and carcinogen-icity in the rat.	47069817
(2006)

Acceptable/Guideline

0, 1000, 4000 or 8000 ppm

M: 0, 39, 159 or 321 mg/kg/day.

F: 0, 56, 220 or 447 mg/kg/day.	NOAEL = 39 for males and 56 for females
mg/kg/day

LOAEL = 159 for males and 220 for females mg/kg/day based mainly on
kidney and urothelial toxicity.

Transitional cell carcinoma in one male and one female.

Gene Mutation

870. 5100	Ames test	47069819 (2006)

Acceptable/Guideline	Not mutagenic at up to 5000 µg/plate.

Gene mutation

870.5100	Ames test	47069820 (2004/2005)

Acceptable/Guideline	Not mutagenic.

Forward mutation

870.5300	V79/HPRT test	47069823 (2006)

Acceptable/Guideline	Negative

Chromo-some abberation

870.5375

479069826

Acceptable/Guideline	No clastogenic effect at up to 1600 µg/ml.

870.5395	Micronucleus test in vivo.	47069829 (2004/2006)

Acceptable/Guideline.	No evidence of clastogenic effect after two doses
up to 2000 mg/kg/(ip)

870.6200a

	Acute neurotoxicity screening battery - rats	47069830 (2006)
Acceptable/Guideline

0, 125, 508 or 2060 mg/kg/day.

	NOAEL = 508  mg/kg/day

LOAEL = 2060  mg/kg/day based on transient urinary staining.

870.6200b

	Subchronic neurotoxicity screening battery - rats	47069831 (2006)

0, 1000, 3000 or 9000 ppm

Acceptable/Guideline

M: 0, 65.2, 193 or 592 mg/kg/day.

F: 0, 84.5, 251 or 748 mg/kg/day.	NOAEL > 592 in males and 748 in
females  mg/kg/day.

LOAEL = Not established since no effects at the highest dose tested.

870.6300

	Developmental neurotoxicity	No study and not required.	N/A

870.7485

	Metabolism and pharmacokinetics

(species)	47069832 (2006) and 47069833 (2006)

Acceptable/Guideline	Absorption, excretion, distribution (very little),
metabolism and identification of metabolites established.

870.7600	Dermal penetration

(species)	No study in laboratory animals 	N/A

	Special studies	No special studies. 	N/A

A.3  Executive Summaries TC \l2 "A.3  Executive Summaries

Note;  The Executive Summaries were prepared by the testing laboratory
and/or the  Bayer Crop Science staff.  They were reviewed by both the
PSD of the United Kingdom and HED toxicologists.  The Executive Summary
is presented together with the comments made by the PSD reviewer and the
HED reviewer in boxes following the Executive Summary.  The HED
Reviewer's comment box includes the classification in HED terminology.
TC \l2 "A.3	Executive Summaries

A.3.1	Subchronic Toxicity TC \l3 "A.3.1	Subchronic Toxicity

	870.3100	90-Day Oral Toxicity - Rat

AE 0001789 (batch number R174, 95.9% w/w purity) was administered
continuously via the diet to separate groups of Wistar rats
(10/sex/group) at dose levels of 0, 1 000, 4 000 and 12 000 ppm
equivalent to 58, 240 and 720 mg/kg/day in males and 70, 281 and 859
mg/kg/day in females, respectively, for at least 90 days.

At 12 000 ppm, there were two female mortalities during week 5 of the
study. One female was found dead on Day 33, clinicals signs observed
prior to death were soiled fur on the muzzle, neck and forelimbs. A
second female was sacrificed for humane reasons on Day 36, clinical
signs observed prior to death were soiled fur on the nose, forelimbs and
anogenital region, wasted appearance and clonic convulsions. This animal
lost 60g in body weight between weeks 4 and 5. In animals surviving to
terminal sacrifice, treatment-related signs consisted of soiled fur in
the anogenital region (observed on one occasion in two males) and hair
loss noted in two males and one female. Absolute body weight in males
was reduced by 5-7% between weeks 5 and 13. Body weight gain was also
reduced at a number of time intervals, the effect being most pronounced
between weeks 4 and 5 where there was a 64% reduction in body weight
gain compared with the controls. In females absolute body weight was
reduced by 6% between weeks 4 and 5. There was an actual loss of body
weight of 0.7 g/day during this interval, however this effect on body
weight was accentuated by a marked loss in body weight of one of the
decedent females. Food consumption in males was reduced by 9 and 13% on
weeks 1 and 5 respectively and by 17% in females during week 5.
Urinalysis revealed sulfonamide-like crystals in the urine of 8/10 males
and 7/8 females. Microscopic examination of the two decedent females,
showed no clear treatment-related cause of death. However, both animals
presented a range of lesions with a number of similarities to those
observed on target organs at terminal sacrifice. At terminal sacrifice,
in male animals, kidney to body weight ratio was increased by 12%,
absolute kidney weight and relative kidney/brain weight were also
slightly increased by 4-5%. In female animals, absolute and relative
kidney weights were increased by 11-13%. At microscopic examination,
effects were seen in the liver and thymus of males and the kidneys,
spleen and urinary bladder of both sexes. In the liver slight
centrilobular hepatocellular hypertrophy was noted in 8/10 males. In the
urinary bladder simple hyperplasia of the urothelium was noted in 2/10
males and 5/8 females. Examination of the kidney revealed a slight
increase in the incidence and/or severity of basophilic tubules and
hyperplasia of collecting ducts and pelvic epithelium, in both sexes. In
addition, metaplasia of the collecting duct epithelium was seen in 1/8
females and brown pigment in the proximal tubules was noted in 2/10
males and 1/8 females examined. In the spleen brown pigment in the red
pulp was seen in animals from all dosage groups but the severity was
marginally greater in a few animals dosed at 12 000 ppm compared to
controls. Examination of the thymus showed a marginal increase in
severity of decreased size of the cortex (involution) in a few males
dosed at 12 000 ppm.

At 4 000 ppm, treatment-related effects were confined to microscopic
findings in the urinary bladder which consisted of simple hyperplasia of
the urothelium in 1/10 males and 1/10 females.

At 1 000 ppm, one male was sacrificed for humane reasons on Day 71
following an accidental trauma. No treatment-related findings were
observed at 1 000 ppm in either sex.

The No Observed Effect Level (NOEL) in the Wistar rat when administered
AE 0001789 in the diet over a 90-day period was 1000 ppm (equivalent to
58 mg/kg/day for males and 70 mg/kg/day for females).

EVALUATION, SUMMARY AND CONCLUSION BY REGULATORY AUTHORITY

Name of authority	Pesticides Safety Directorate, UK

Reviewer’s Comments	Reliability rating: Totally reliable

The study (Mc Elligott A. ; 2003, SA 02352) is compliant with OECD 408
(1998) with the following deviation:

The guideline specifies that histopathology of the brain should include
investigation of ‘representative regions including cerebrum,
cerebellum and medulla pons’.  The study report states only that the
brain was investigated histopathologically. The Notifier states that
‘Routinely, three transverse sections are made for histopathological
examination:

1.	the first section at the level of optic chiasm including the basal
ganglia, septum, cortex, anterior hypothalamus

2.	the second section at the level of hippocampus containing the cortex
and brain stem at the transition of diencephalon to mesencephalon

3.	the third section containing the cerebellum and brain stem (medulla
oblongata)’

	It is not clear how the conclusion was drawn that the crystals in the
urine were ‘sulphonamide like’ using these methods given.  The
Notifier states that ‘These crystals have been identified visually
under the microscope based on the specific shape of such crystals. These
crystals look like shocks of wheat or radially-striated spherules’

At l2000 ppm in males, kidney to bodyweight ratio was increased by 12%
(p<0.01) compared to controls. Absolute kidney weight and relative
kidney/brain weights were also slightly increased by 4-5%. In females
absolute and relative kidney weights were increased by 11-12%, although
none of these increases were statistically significant.

Although some microscopic changes were observed in the liver and spleen
there were no correlated clinical chemical or haematological findings.

Urine analysis revealed an apparent increase in urine volume in females.

	Males

Parameter	Dose level (ppm)

0	1000	4000	12000

	Kidney weight

(g)	M	3.01	3.07	3.00	3.16

	Kidney weight

(% body weight)

0.585	0.584	0.592	0.657*

	Kidney weight

(% brain weight)

142.512	145.006	141.698	148.282

	urine volume

(ml)

3.9	4.7	3.6	4.6

	Females

Parameter	Dose level (ppm)

0	1000	4000	12000

	Kidney weight

(g)	F	1.74	1.81	1.76	1.93

	Kidney weight

(% body weight)

0.630	0.630	0.629	0.697

	Kidney weight

(% brain weight)

88.034	92.363	90.922	99.777

	urine volume

(ml)

2.4	2.4	2.2	5.0

Conclusions	The results of this study indicate that cyprosulfamide is of
relatively low toxicity in the rat. The main target organs were with the
kidney and urinary bladder, with effects also seen on the liver (males
only), thymus (males only) and spleen.

The changes seen in the kidney (basophilic tubules and hyperplasia of
collecting ducts and pelvic epithelium) and bladder (simple hyperplasia
of the urothelium) are consistent with a local inflammatory response
(hyperplasia) due to the presence ‘sulphonamide-like” crystals which
were observed in the urine from rats treated at doses >4000 ppm.

In the spleen, an increase in the severity of brown pigment in the red
pulp was seen at the top dose 12000 ppm compared to controls. The
effects was considered by the authors to be a consequence of an increase
in the turnover of red cells in affected animals, although
haematological analysis didn’t support this theory.

In the absence of any other effects the changes seen in the thymus in a
few top dose males (decreased size of the cortex (involution) was likely
a result of stress rather than an effect on the immune system

	A NOAEL of 1000 ppm (equivalent to mean achieved dietary intakes of 58
and 70 mg/kg bw/d in males and females respectively) based on
hyperplasia of the urothelium in the urinary bladder at 4000 ppm with
‘Sulphonamide-like’ crystals also seen in the urine at this dose.

HED Reviewer comments:

The study is classified as ACCEPTABLE/GUIDELINE and satisfies the
requirement for a series 870.3100 subchronic toxicity study in the rat.
HED concurs with the assignment of the NOAEL and LOAEL.

	870.3100	90-Day Oral Toxicity - Mouse (MRID No.: 47069809)

AE 0001789 (Batch number R174, 95.9 % w/w purity), was administered
continuously via the diet to groups of C57BL/6 mice (10/sex/group) at
concentrations of 0, 500, 2000 and 7000 ppm for at least 90 days
(equating approximately to 0, 79, 321 and 1110 mg/kg/day in males and 0,
95, 398 and 1297 mg/kg/day in females).

At 7 000 ppm, the only treatment-related finding was an increased
incidence and severity of lymphocytolysis in the thymus of female mice
only.

At 2 000 and 500 ppm, no treatment-related findings were observed in
either sex.

The NOEL of AE 0001789 in this study was 2 000 ppm in females (equating
398 mg/kg/day) and 7 000 ppm in males (equating to 1 110 mg/kg/day).

EVALUATION, SUMMARY AND CONCLUSION BY REGULATORY AUTHORITY

Name of authority	Pesticides Safety Directorate, UK

Reviewer’s Comments	Reliability rating: Totally reliable

The study (Kennel P., 2003, SA 03005) is compliant with OECD 408 (1998)
with the following deviation:

The guideline specifies that histopathology of the brain should include
investigation of ‘representative regions including cerebrum,
cerebellum and medulla pons’.  The study report states only that the
brain was investigated histopathologically. The Notifier states that
‘Routinely, three transverse sections are made for histopathological
examination:

1.	the first section at the level of optic chiasm including the basal
ganglia, septum, cortex, anterior hypothalamus

2.	the second section at the level of hippocampus containing the cortex
and brain stem at the transition of diencephalon to mesencephalon

3.	the third section containing the cerebellum and brain stem (medulla
oblongata)’

The epididymides and ovaries were not weighed, as specified in OECD 408,
however this is not considered to affect the integrity of the study in
the absence of effects on these organs in other studies in the mouse or
in other species

The only notable finding in this 90 day mouse study was at the top dose
with a treatment related increase in the incidence and severity of
lymphocytolysis in the thymus. There were no other treatment-related
findings.

Conclusions	The results of this study indicate that cyprosulfamide is of
low toxicity in the mouse. The only finding was increase in the
incidence and severity of lymphocytolysis in the thymus in high dose
females, which was considered by the Notifier to be stress related
change, although there was little evidence of any effects which could
cause stress in these animals. There was no evidence of any signs of
toxicity including an absence of any clinical signs, nor any effects on
the immune system.

	Based on an absence of effects in males the NOAEL was 7000 ppm
(equivalent to a mean achieved dietary intake 1110 mg/kg bw/day) and in
females 2000 ppm (equivalent to a mean achieved dietary intake 398 mg/kg
bw/day) based on the effects seen in the thymus.

HED Reviewer Comments:

Classification/Acceptable Guideline.   The study serves as a dose range
finding study for the definitive carcinogenicity study in mice.

	870.3150	90-Day Oral Toxicity - Dog (MRID No.: 47069811)

Technical grade AE 0001789 (Batch number 08466/0013, 97.4% purity) was
administered in the diet to Beagle dogs (4/sex/dose) at dose levels of 0
(concurrent vehicle control), 2 500, 7 500 and 15 000 ppm for a
period of at least 90 days (equivalent to 75, 221 and 416 mg/kg/day for
the males and 79, 221 and 341 for the females, respectively). Clinical
observations were conducted twice daily, food consumption was measured
daily and body weights were taken weekly. Clinical chemistry,
haematology and urinalysis were performed on all animals once prior to
administration of the test substance and during study weeks five, nine
and twelve. Ophthalmic examinations were performed pre-exposure and
pre-sacrifice. A gross necropsy was performed, organ weights were taken
and tissues were examined microscopically.

At 15 000 ppm: One female was sacrificed for humane reasons. This death
was considered treatment-related due to the kidney calculi and
abscessation. There was a body weight loss in the females and no effect
on body weight for the males. There was a reduction of about 15%
compared to controls in food consumption for both sexes. Elevated urea
nitrogen compared to pre-test values was observed in males (from 92 to
169 %) and in females (from 177 to 554%); elevated serum creatinine was
also observed in males and females (from 44 to 60% and 44 to 210%,
respectively). Elevated serum phosphorus was only noticed in females
(from 49% compared to pre-test values at termination). Treatment-related
urinalysis findings were: elevated urine volume in males, a decrease in
urine specific gravity in males and females, decreased pH in females.

Calculi in the renal pelvis were seen in 3/4 males and all females,
renal pelvis dilatation was observed in one male and one female. Calculi
were observed in the ureter and urinary bladder of one male and dilated
ureters were seen in one female. Correlated microscopic changes of the
urinary tract were inflammatory changes, renal pelvis epithelium
hyperplasia and necrosis and associated changes in the ureters and
urinary bladder of males and females.

At 7 500 ppm: The only treatment-related finding was a decrease in
urine specific gravity in females.

At 2 500 ppm: No treatment-related effects were observed.

The No Observed Adverse Effect Level for this study was 7 500 ppm
(equilavent to 221 mg/kg/day for males and females) as there were no
microscopic pathology observations to correlate with the decrease in
urine specific gravity in the 7 500 ppm females.

EVALUATION, SUMMARY AND CONCLUSION BY REGULATORY AUTHORITY

Name of authority	Pesticides Safety Directorate, UK

Reviewer’s Comments	Reliability rating: Totally reliable

	The study (M-263605-01-1) is fully compliant with OECD 409 (1998).
Although reticulocyte counts are listed under haematological
investigations in the study no results are reported, however red cell
morphology has been assessed

	There was a treatment-related decrease in bodyweights for the top dose
females (a loss in weight of -874 g for the high-dose group during the
study), top dose males gained weight but only 30% of the gain seen in
males. Food consumption was decreased in both sexes at the top dose.

Bodyweight changes in 90 day dog study;

	Dose	Bodyweight (g)	weight

	ppm	Day 0	week 1	week2	week 3	week 7	week 12	change (kg)

	male

	0	7943	8342	8529	8376	8590	8876	932.7

	2500	7851	8196	8482	8426	8827	8921	1070

	7500	8471	8848	9239	9092	9383	9721	1250.5

	15000	8052	8201	8260	8175	8392	8329	277.2

	Female

	0	7046	7218	7268	7219	7437	7504	458

	2500	7110	7294	7495	7314	7460	7515	404.5

	7500	7003	7206	7285	7155	7464	7596	592.8

	15000	6661	6881	6817	6516	6566	5786	-874.2

Food consumption in 90 day dog study;

Dose group	Food consumption (kg)

	ppm	Males 	Females

	0	24.7	19.3

	2500	23.9	17.9

	7500	24.9	19.6

	15000	21.2	16.5

	A very high WBC (57.8 x103/mm3) in one early death high-dose group
female (VG3102) on Study Day 35 day was thought to be due to kidney
changes described microscopically (abscess). Mild treatment related
anaemia was evident with statistically significant decrease in
erythrocyte counts, haemoglobin, and hematocrit in the high-dose group
females at termination. Some haematological changes (MCV, MCHC, RDW, and
HDW) were also noted at 7500 ppm although these were fairly minor in
magnitude. Some statistically significant but small changes in
haematological parameters were noted in males mostly at the top dose
changes (MCV, MCHC, RDW, and HDW). In males at doses > 7500 ppm there
was a clear increase in % reticulocytes which correlated to the pattern
of mild anaemia.

	Haematological 	Dose Group (ppm)

	Findings	males

0	2500	7500	15000

	WBCs (103/mm3)	day -7	9.3	7.9	8.5	9.9

day 35	9.7	8.5	10.0	10.9

day 63	9.8	8.2	9.9	11.4

day 84	9.6	7.7	10.9	8.2

	RBCs (106/mm3)	day -7	6.92	7.01	7.16	7.02

day 35	6.89	6.69	6.99	6.63

day 63	7.00	7.34	6.91	6.30

day 84

	7.17

	7.20

	6.90

	6.46

	Hgb (g/dl)	day -7	16.1	16.3	17.0	16.5

day 35	16.5	15.8	17.0	16.0

day 63	16.8	17.4	16.8	15.6

day 84	17.2	17.1	17.0	16.2

	Hct (%)	day -7	45.8	46.1	48.0	47.3

day 35	46.8	45.0	48.0	46.6

day 63	47.5	49.2	47.0	46.2

day 84	48.9	48.8	48.4	48.7

	MCHC (g/dl)	day -7	35.1	35.4	35.6	34.7

day 35	35.3	35.2	35.4	34.4*

day 63	35.3	35.3	35.8	33.8*

day 84	35.2	35.2	35.2	33.4*

	MCV (µm3)	day -7	66.2	65.8	67.0	67.5

day 35	67.9	67.5	68.7	70.4

day 63	67.9	67.4	68.0	73.2*

day 84	68.2	68.0	70.1	75.3*

	RDW (%)	day -7	14.1	13.9	13.8	14.2

day 35	12.6	12.8	12.7	12.5

day 63	12.8	12.8	12.4	13.7

day 84	12.9	12.8	13.3	11.9*

	HDW (%)	day -7	1.59	1.67	1.67	1.61

day 35	1.60	1.61	1.61	1.58

day 63	1.66	1.66	1.62	1.93*

day 84	1.73	1.71	1.84*	1.57*

	Reticulocytes (%)	day -7	0.5	0.6	0.9	0.7

day 35	0.7	0.9	1.3	1.0

day 63	0.9	0.9	1.0	2.9*

day 84	1.0	1.0	2.0*	1.6

	Haematological	FEmales

	Findings	0	2500	7500	15000

	WBCs (103/mm3)	day -7	7.7	12.3	8.3	8.2

day 35	8.7	10.2	10.0	20.9

day 63	8.7	9.8	10.2	10.3

day 84	7.9	10.2	9.3	5.7

	RBCs (106/mm3)	day -7	7.42	6.98	7.26	7.33

day 35	6.84	6.37	7.12	6.12

day 63	6.75	6.61	6.76	6.04

day 84	7.05	6.51	6.59	5.18*

	Hgb (g/dl)	day -7	17.4	16.3	16.9	17.2

day 35	16.5	15.2	16.7	14.5

day 63	16.1	15.7	16.0	14.6

day 84	16.9	15.7	15.9	12.5*

	Hct (%)	day -7	49.3	47.2	48.7	49.2

day 35	46.3	42.8	48.8	41.9

day 63	45.3	44.8	46.7	43.1

day 84	48.1	44.9	46.2	38.0*

	MCHC (g/dl)	day -7	35.4	34.6	34.7	35.0

day 35	35.6	35.5	34.3*	34.7

day 63	35.6	35.0	34.3*	33.8*

day 84	35.1	35.0	34.4*	32.9*

	MCV (µm3)	day -7	66.4	67.4	67.1	67.1

day 35	67.7	67.2	68.5	68.5

day 63	67.2	67.8	69.1	71.5

day 84	68.2	68.9	70.2*	73.4*

	RDW (%)	day -7	13.5	13.3	13.1	13.1

day 35	12.9	12.4	12.3	12.4

day 63	12.1	12.7	12.2	12.5

day 84	12.3	12.4	12.2	11.7

	HDW (%)	day -7	1.57	1.50	1.52	1.43

day 35	1.72	1.55	1.55	1.58

day 63	1.66	1.68	1.59	1.73

day 84	1.57	1.58	1.59	1.64

	Reticulocytes (%)	day -7	0.5	0.7	0.7	0.3

day 35	1.0	0.7	0.9	0.9

day 63	0.7	0.9	0.7	1.4

day 84	0.7	0.7	1.0	0.9

	Urinalysis revealed a number of treatment related effects at 15000 ppm;
elevated urine volume in males, decreased specific gravity in males and
females and a decrease in pH in females. The only effect evident at 7500
ppm was a decreased specific gravity in females.

	Urinalysis	Dose Group (ppm)

	Findings	males

0	2500	7500	15000

	pH	day 0	8.0	7.5	7.6	8.3

1 month	8.8	8.8	8.4	7.6

2 month	7.6	9.0	8.1	7.5

term.	7.4	7.5	7.9	7.6

	Specific Gravity	day 0	1.028	1.020	1.012	1.020

1 month	1.022	1.021	1.014	1.015

2 month	1.018	1.033	1.019	1.012

term.	1.028	1.034	1.010	1.004*

	Urine volume (ml)	day 0	171.5	181.0	135.0	111.3

1 month	223.3	383.8	203.3	840.5

2 month	373.3	295.3	266.5	582.0

term.	137.0	298.3	185.8	815.3

	Urinalysis	FEmales

	Findings	0	2500	7500	15000

	pH	day 0	7.9	7.0	7.9	8.8

1 month	8.4	7.8	7.6	6.2*

2 month	9.0	7.6	8.5*	6.3*

term.	8.0	7.3	6.9	7.2

	Specific Gravity	day 0	1.030	1.022	1.014	1.034

1 month	1.021	1.027	1.021	1.011

2 month	1.027	1.031	1.032	1.011

term.	1.031	1.018	1.008*	1.009*

	Urine volume (ml)	day 0	102.5	151.0	173.0*	61.8

1 month	303.0	158.5	271.8	303.7

2 month	263.8	204.5	183.5	428.7

term.	128.8	194.0	217.5	339.7

	Gross pathological examination revealed a small thymus in three of the
four females at the top dose (mean absolute weight control 8.529g cf.
15000ppm 3.936g; mean relative (to bw) weight control 0.112% cf.
15000ppm 0.102g) these differences were considered to be due to the
reduced overall bodyweight in the top dose group.

	There was a statistically significant decrease in absolute testes
weight at the top dose, and a non-statistically significant decrease in
relative (to bodyweight) testes weight. There were no other related
findings.

	Absolute and relative (to bw) testes weight in 90 day dog study

Dose group	Testes weight

	ppm	absolute (g) 	relative (%)

	0	12.983	0.146

	2500	13.668	0.152

	7500	14.036	0.147

	15000	9.964*	0.123

*= p<0.05

Conclusions	The top dose in this 90 day study causing a weight loss in
females and the death (in-extremis) of a female probably due to kidney
calculi and resultant abscessation.   There was also a reduction in food
consumption. Overall bodyweight gain was reduced in males at the top
dose (ca 70% lower than controls) there was also a slight reduction in
food consumption.

The urinary system was found to be the main target of toxicity as is
also the case in short-term toxicity studies in the rat and mouse.
Inflammatory changes were evident in the renal pelvis, epithelial
hyperplasia and necrosis, and associated changes in the ureters
(dilatation) and urinary bladder. Gross necropsy revealed the probable
cause of these changes with calculi observed in the kidneys, ureters and
urinary bladder. Clinical chemistry revealed some related findings at
15000 ppm; elevated urea nitrogen in both sexes, elevated serum
creatinine in both sexes and elevated serum phosphorus in females.
Urinalysis at 15000 ppm also revealed elevated urine volume in males,
decrease in urine specific gravity in both sexes and decreased pH in
females.  A decrease in urine specific gravity was also seen in females
at 7500 ppm, although without histopathological correlate.

In addition cyprosulfamide appeared to cause mild anaemia in top dose
females characterised by statistically significant decrease in
erythrocyte counts, haemoglobin, and hematocrit. There were also some
slight changes in haematological parameters at 7500 ppm, although minor
in nature. There was some evidence for a similar effect in males
although this was marginal. There were no correlated microscopic
findings.

A NOAEL of 7500 ppm (equivalent to mean achieved dietary intakes of
221 mg/kg bw/day in males and females) can be determined for this
study, based on the clear effects on the urinary system, bodyweight
changes and mild anaemia at the top dose.  The effects observed at 7500
ppm were not biologically significant and not considered adverse.

I

HED Reviewer Comments:

Classification:  Acceptable/Guideline.  The study satisfies the
guideline requirement for a series 870.3150 subchronic toxicity study in
dogs.  HED concurs with the assignment of the NOAEL and LOAEL.

	870.3200	21/28-Day Dermal Toxicity – Rat  -

No study available and not required.

	870.3465	90-Day Inhalation – Rat

No study available and not required.

A.3.2	Prenatal Developmental Toxicity TC \l3 "A.3.2	Prenatal
Developmental Toxicity

	870.3700a Prenatal Developmental Toxicity Study - Rat (MRID No.:
47069814)

In a developmental toxicity study AE 0001789 (batch number 08466/0013,
97.4% w/w purity) was administered daily by gavage from gestation day
(GD) 6 to 20 to groups of 25 pregnant Sprague-Dawley female rats per
dose-group. The doses given were 0, 60, 250 and 1000 mg/kg/day in
suspension in aqueous solution of 0.5% methylcellulose 400.  Clinical
observations were recorded daily and body weights were recorded for all
females on GD 0, 6, 8, 10, 12, 14, 16, 18 and 21. Food consumption was
also measured for all the females during the intervals GD l-6, 6-8,
8-10, 10-12, 12-14, 14-16, 16-18 and 18-21. At scheduled sacrifice, on
GD 21, the gravid uterine weight was recorded and the dams were
evaluated for number of corpora lutea, number and status of
implantations (resorptions, dead and live fœtuses). In addition, the
kidney and the urinary bladder of all dams were examined for signs of
gritty material. These organs were retained from all females, the kidney
was examined histopathologically at all dose levels, whilst the urinary
bladder was examined at 1000 mg/kg/day and from the control group only.
Live fœtuses were removed from the uteri, counted, weighed, sexed and
examined externally. Approximately half of the live fœtuses from each
litter were fixed in Bouin's solution and subsequently dissected for
internal examination. The remaining half were eviscerated, fixed in
absolute ethanol and stained according to a modification of the Tyl and
Marr technique for skeletal examination of bone and cartilage.

There were no mortalities or treatment-related signs in any dose group.

At 1000 mg/kg/day, body weight change was reduced in dams by 32% between
GD 6 to 8, thereafter, body weight change was comparable to the
controls. Maternal corrected body weight change was reduced by 11%
compared to controls. At autopsy, one female was noted with yellow
sediment in both kidneys and prominent lobulation of the liver, one
female had an enlarged liver with white foci on the left lobe and one
female had a white area on the left lobe. At microscopic examination,
one female was found to have a moderate bilateral papillary necrosis in
the kidney, associated with mixed cell infiltrate. The change correlated
with the macroscopic finding of yellow bilateral sediment in the kidney
of this female. No litter parameters were affected, including the number
of live fœtuses, early and late resorptions, fœtal death status,
percentage of male fœtuses and mean body weights. At external, visceral
and skeletal fœtal examination, no treatment-related findings were
observed.

At 250 and 60 mg/kg/day, there were no treatment-related maternal,
litter based or fœtal findings at either dosage.

In conclusion, a dose level of 1000 mg/kg/day AE 0001789 caused toxic
effects to the dams and was a No Observed Effect Level (NOEL) in terms
of fœtal development. A dose level of 250 mg/kg/day was the NOEL for
maternal toxicity.

EVALUATION, SUMMARY AND CONCLUSION BY REGULATORY AUTHORITY

Name of authority	Pesticides Safety Directorate, UK

Reviewer’s Comments	Reliability rating: totally reliable

The study (M-261462-01-2) is fully compliant with OECD 414 (2001).
Dosing from Day 6-20 of gestation is acceptable.

Maternal bodyweight effects were clearly seen at the top dose.

	Maternal weight (g) Pregnant females	Dose level (mg/kg bw/d)

0	60	250	1000

	Gestation day 0	269.2	269.4	269.2	269.7

	Gestation day 6	305.8	305.0	302.9	303.2

	Gestation day 8	314.2	312.6	309.9	309.0

	Gestation day 10	324.5	322.5	319.4	321.1

	Gestation day 12	335.0	333.0	330.0	330.6

	Gestation day 14	342.0	342.1	339.2	340.4

	Gestation day 16	359.4	358.3	355.9	358.4

	Gestation day 18	387.9	385.6	383.4	386.4

	Gestation day 21	436.5	432.1	432.2	434.8

	External Observations: There were three fetuses observed with
malformations, two at 250 mg/kg bw/day and one at 60 mg/kg bw/day. At
250 mg/kg/day, one fetus had an umbilical hernia and one fetus had
anasarca, whilst at 60 mg/kg/day one fetus had a cleft palate.  In
isolation and in the absence of a dose response, these malformations
were considered to be incidental.

Visceral Observations: At 1000 mg/kg bw/day, one fetus had the
malformation retinal fold associated with a small lens, whilst a second
fetus had the malformation right-sided aortic arch and pulmonary trunk.
As these two malformations occurred in isolation, they were considered
to be incidental. In addition, the incidence of the malformation absent
renal papilla {unilateral} was marginally higher in terms of both mean
percentage of fetuses affected and percentage of litters affected
compared with the control group. The incidence was not statistically
significant for either parameter, when compared to the control group.
The incidence in terms of litters, and was just within the historical
control range, although over in terms of fetuses. Overall it is
considered that it is unlikely that this finding was treatment related.

	Observation: Absent renal papilla (unilateral)	Dose level (mg/kg
bw/day)	Historical Control Range

0	60	250	1000

Mean % fetuses affected	0.6	0.0	0.6	1.9	0.0-1.0

	Mean % litters affected	4.0	0.0	4.0	8.0	0.0-8.3

	Skeletal Observations: Two fetuses with malformations were noted, one
occurred at 1000 mg/kg bw/day and comprised of hemivertebra in the atlas
vertebra and malpositioned axis vertebra. The second malformation
occurred at 60 mg/kg bw/day, where one fetus had a small and split
palatine (the same fetus which had a cleft palate at the external
observation). In isolation, these malformations are considered to be
incidental. The following two variants were found at a higher incidence
at 1000 mg/kg bw/day; incomplete parietal ossification and incomplete
interparietal ossification. In addition, the incidence of 13th costal
cartilage absent (unilateral/bilateral), classified as a variant, was
marginally higher at 1000 mg/kg bw/day, compared with the controls.
However, as in all three cases the incidence was found not to be
statistically significantly different from the control group at both the
fetal and litter level, these findings were considered to be incidental.

Conclusions	No evidence of teratogenicity was seen in this study.

A NOAEL for maternal toxicity of 250 mg/kg bw/day can be determined,
based on the reduced weight gain and effects on the kidneys seen at the
top dose level of 1000 mg/kg bw/day.  A NOEL for developmental toxicity
of 1000 mg/kg bw/day can be based on the absence of any affect on fetal
parameters.

HED Reviewer Comments:

Classification:  Acceptable/Guideline.  The study satisfied the
guideline requirement for a serried 870.3700 developmental toxicity
study in rats.   HED concurs with the assignment of the NOAEL and LOAEL.

	870.3700b Prenatal Developmental Toxicity Study - Rabbit

First study (MRID No.: 47069812)

Twenty-five time-mated female New Zealand White Rabbits per group were
exposed to AE 0001789, a herbicide safener belonging to the reverse
sulfamide family, (batch number 08466/0013, a white powder, 97.4% w/w
purity) by gavage from Gestation Day (GD) 6 to 28. The doses given were
0, 50, 125 and 500 mg/kg/day in suspension in aqueous solution of 0.5%
methylcellulose 400. The high dose level of 500 mg/kg/day was
terminated prior to commencement of scheduled autopsy, due to the high
mortality rate observed at this dosage.

Maternal body weights were recorded for all surviving females on GD 3,
6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26 and 29. Food consumption was
measured for all surviving females during the intervals GD 3-6, 6-8,
8-10, 10-14, 14-18, 18-22, 22-26 and 26-29. Clinical observations were
recorded daily. At scheduled sacrifice, on GD 29, the gravid uterine
weight and the number of ribs (in pregnant females) were recorded and
the dams were evaluated for number of corpora lutea, number and status
of implantations (resorptions, dead and live fœtuses). The urinary
bladder and kidney were retained from all females at 50 and
125 mg/kg/day and the control group, but were examined microscopically
at 125 mg/kg/day and from the control group only. Live fœtuses were
removed from the uteri, counted, weighed and examined externally. The
heads of fœtuses from approximately half of each litter were immersed
in Bouin’s fluid and the internal structures examined after fixation.
The bodies of all fœtuses were dissected for soft tissue anomalies and
sexed. Fœtuses were eviscerated, skinned and fixed in absolute ethanol
before staining. A modification of the Staples and Schnell technique was
applied and a subsequent skeletal examination was performed.

At 500 mg/kg/day

Between GD 9 and 20, seven of 25 dams died; four were killed for humane
reasons and three were found dead. The principal clinical signs observed
were yellow sediment in the urine in all animals, associated with red
traces on the cage tray in five cases. All animals showed a marked body
weight loss, associated with very poor food consumption prior to death.
At autopsy, the principal target organs were the kidney and urinary
bladder, where yellow sediments were noted in the kidney of 4/7 females
and in the urinary bladder of 2/7 females. In addition, a marbled
pattern was observed on the kidney in 2/7 cases and red contents in the
urinary bladder in 1/7 females. As this dose level had clearly exceeded
a Maximum Tolerated Dose (MTD), treatment at 500 mg/kg/day was
terminated. Surviving females were killed prior to scheduled autopsy
with no macroscopic examination performed and no tissues taken. Of the
females surviving until termination of this dose level,
treatment-related clinical signs consisted of yellow sediment in the
urine in 3/18 dams and few faeces in 4/18 dams. Two females showed a
significant body weight loss of between 0.21 and 0.29 kg, and poor food
consumption prior to necropsy.

At 125 mg/kg/day

At this dosage, treatment-related clinical signs consisted of yellow
sediment in the urine in 2/25 dams. No effect on mean body weight
change, mean corrected body weight change or mean food consumption was
noted. At autopsy, yellow sediment in the kidney was noted in 1/25
females. No treatment-related changes were detected at the microscopic
examination of the kidney or urinary bladder. As a consequence, the
macroscopic finding of marbled pattern on the kidney in the decedent
female and yellow sediment in the kidney in one female at scheduled
sacrifice were regarded as not adverse findings. No litter parameters
were affected at this dosage. There were no external, visceral or
skeletal findings at the fœtal evaluation which were considered to be
treatment-related at this dosage.

At 50 mg/kg/day

There were no treatment-related clinical signs, and mean body weight
change, mean corrected body weight change and mean food consumption were
unaffected by treatment. At autopsy, 1/25 females had a kidney with a
marbled pattern appearance, but in the absence of microscopic findings
in the kidney at 125 mg/kg/day, this finding was considered to be not an
adverse finding. No treatment-related effects on litter parameters were
observed at this dosage. There were no external, visceral or skeletal
findings at the fœtal evaluation which were considered to be
treatment-related at this dosage.

In conclusion, a dose level of 500 mg/kg/day clearly exceeded a Maximum
Tolerated Dose (MTD) in the pregnant rabbit. A dose level of
125 mg/kg/day was a No Observed Adverse Effect Level (NOAEL) in the dam
due to the findings observed in the kidney, whilst being a No Observed
Effect Level (NOEL) in the foetus, in the New Zealand White Rabbit.

EVALUATION, SUMMARY AND CONCLUSION BY REGULATORY AUTHORITY

Name of authority	Pesticides Safety Directorate, UK

Reviewer’s Comments	Reliability rating: totally reliable

The study (M-261462-01-2) is fully compliant with OECD 414 (2001).
Dosing from Day 6-28 of gestation is acceptable.

Between GD 9 and 20, 7/25 dams died; four were killed for humane reasons
and three were found dead.  The principal clinical signs observed were
yellow sediment in the urine in all animals, associated with red traces
on the cage tray in five cases.  All animals had few/no faeces on one or
more occasion and one female had reduced motor activity, tremors and was
warm to touch on the day of death.  All animals showed a marked body
weight loss, associated with very poor food consumption prior to death.
At autopsy, the principal target organs were the kidney and urinary
bladder, where yellow sediments were noted in the kidney of 4/7 females
and in the urinary bladder of 2/7 females.  In addition, a marbled
pattern was observed on the kidney in 2/7 cases and red contents in the
urinary bladder in 1/7 females.  As this dose level had clearly exceeded
a Maximum Tolerated Dose (MTD), treatment at 500 mg/kg bw/day was
terminated. Surviving females were killed prior to scheduled autopsy
with no macroscopic examination performed and no tissues taken.

At 125 mg/kg/day, one female was found dead on GD 19, shortly after
dosing, prior adverse findings. At autopsy, red contents were found in
the trachea, all lobes of the lung were discoloured and a marbled
pattern was noted on the kidney.  This animal was considered to have
died due to gavage error.

At 50 mg/kg/day, one female was found dead on GD 15; this animal had
reduced activity and slow respiration on GD 15.  At autopsy, macroscopic
findings consisted of red contents in the trachea, and haemorrhaging and
discoloration in the lungs.  The cause of death was judged to have been
gavage error.

At 125 mg/kg/day, yellow sediment was observed in the urine of 2/25
dams.

At autopsy, yellow sediment in the kidney was noted in 1/25 females at
125 mg/kg bw/day, whilst the decedent female at 125 mg/kg bw/day and
1/25 females at 50 mg/kg bw/day had a kidney with a marbled pattern
appearance. There were no microscopic findings in this organ at 125
mg/kg/day.  These changes were clearly treatment related although not
adverse.

Microscopic examination of the urinary bladder and kidney at 125 mg/kg
bw/day revealed no treatment-related findings were noted.

Maternal bodyweight effects were clearly seen at the 500 mg/kg bw/day
but not at 50 or 125 mg/kg bw/day.

	Maternal weight (kg) Pregnant females	Dose level (mg/kg bw/d)

0	50	125	500

	Gestation day 3	3.48	3.52	3.46	not applicable

	Gestation day 6	3.48	3.50	3.46	not applicable

	Gestation day 8	3.49	3.50	3.46	not applicable

	Gestation day 10	3.51	3.51	3.48	not applicable

	Gestation day 12	3.52	3.53	3.51	not applicable

	Gestation day 14	3.55	3.55	3.54	not applicable

	Gestation day 16	3.59	3.57	3.58	not applicable

	Gestation day 18	3.61	3.60	3.60	not applicable

	Gestation day 20	3.62	3.62	3.64	not applicable

	Gestation day 22	3.64	3.65	3.67	not applicable

	External Observations: There were no treatment related findings.

Visceral Observations.     There were no treatment related findings

Skeletal Observations: The incidence variants were higher in both
treated groups or at 125 mg/kg bw/day alone, compared to the controls,
in terms of both fetal and litter parameters: 13th thoracic rib(s)
(unilateral/bilateral) short, 13th thoracic rib(s)
(unilateral/bilateral) (including short) detached, hyoid centrum
incomplete ossification or unossified, hindpaws 5th middle or 5th distal
phalanges unossified, less than 15 caudal vertebrae. These findings were
all within the historical control range for both fetal and litter
parameters at the higher dose level of 125 mg/kg/day, and there was no
clear dose response the relationship to treatment was considered
equivocal. The apparent effects seen on the 5th middle or 5th distal
phalanges are difficult to dismiss as not being a possible treatment
related effect based on the results of this study alone. However taken
in the context of the second developmental study in the rabbit these
changes are likely incidental.

	Observation: 	Dose level (mg/kg bw/day)	Historical Control Range

0	50	125	500

13th thoracic rib(s) (unilateral/bilateral)

	Mean % fetuses affected	12.4	18.1	16.6	na	11.5-19.7

	Mean % litters affected	47.6	82.6	70.8	na	58.3-78.3

	13th thoracic rib(s) (unilateral/bilateral) (inc. short detached)

	Mean % fetuses affected	7.7	11.9	9.5	na	5.1-12.7

	Mean % litters affected	23.8	69.6	45.8	na	33.3-58.3

	hyoid centrum incomplete ossification or unossified

	Mean % fetuses affected	17.9	25.2	21.1	na	15.4-26.2

	Mean % litters affected	28.6	60.9	54.2	na	37.5-63.6

	hindpaws 5th middle or 5th distal phalanges unossified

	Mean % fetuses affected	0.5	0.5	1.5	na	0.0-1.7

	Mean % litters affected	4.8	4.3	12.5	na	0.0-13.6

	less than 15 caudal vertebrae

	Mean % fetuses affected	0.5	2.9	1.8	na	0.0-2.3

	Mean % litters affected	4.8	26.1	16.7	na	0.0-20.3

	na not applicable

	Conclusions	No evidence of teratogenicity was seen in this study.

A NOAEL for maternal toxicity of 125 mg/kg bw/day can be determined,
based on the marked toxicity seen at 500 mg/kg bw/day.  A NOEL for
developmental toxicity of 125 mg/kg bw/day can be based on the absence
of any affect on fetal parameters.

HED Reviewer Comments:

Classification: Acceptable/Guideline.  This study satisfies the
guideline requirement for a series 870.3700 developmental toxicity study
in rabbits when taken together with the follow up study (MRID No.:
47069813).

Second Study (MRID No.: 87069813).

Twenty-five time-mated female New Zealand White Rabbits per group were
exposed to AE 0001789, a herbicide safener belonging to the reverse
sulfamide family, (batch number 08466/0013, a white powder, 97.4% w/w
purity) by gavage from Gestation Day (GD) 6 to 28. The doses given were
0 and 250 mg/kg/day in suspension in aqueous solution of 0.5%
methylcellulose 400. This study was conducted as a complementary study
to SA 03349 developmental toxicity study in the rabbit with the same
compound, where the high dose level of 500 mg/kg/day was found to be too
toxic to the dams.

Maternal body weights were recorded for all surviving females on GD 3,
6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26 and 29. Food consumption was
measured for all surviving females during the intervals GD 3-6, 6-8,
8-10, 10-14, 14-18, 18-22, 22-26 and 26-29. Clinical observations were
recorded daily. At scheduled sacrifice, on GD 29, the gravid uterine
weight and the number of ribs were recorded and the dams were evaluated
for number of corpora lutea, number and status of implantations
(resorptions, dead and live fœtuses). The urinary bladder and kidney
were retained from all females but not histologically examined. Live
fœtuses were removed from the uteri, counted, weighed and examined
externally. The heads of fœtuses from approximately half of each litter
were immersed in Bouin’s fluid and the internal structures examined
after fixation. The bodies of all fœtuses were dissected for soft
tissue anomalies and sexed. Fœtuses were eviscerated, skinned and fixed
in absolute ethanol before staining. A modification of the Staples and
Schnell technique was applied and a subsequent skeletal examination was
performed.

In the treatment group at 250 mg/kg/day, two dams were sent to necropsy
on GD 28 after aborting. Clinical signs for the first female consisted
of few/mucoid faeces on several occasions, yellow sediment in the urine
on GD 24-26 and 28, and anogenital soiling, red traces on tray and
placental tissue on tray on GD 28. Clinical signs for the second female
included no/few faeces on several occasions, yellow sediment in the
urine on GD 23-28, red traces on tray on GD 26-28 and
placental/fœtoplacental tissue on tray on GD 28. Between GD 10 and 28
the first female lost 0.49 kg, whilst between GD 20 and 28 the second
female lost 0.57 kg in body weight. In both cases, the body weight loss
was associated with a marked reduction in food consumption. No
treatment-related findings were observed in either female at the
macroscopic examination.

In surviving females, 5/23 animals had yellow sediment in the urine on
one or more occasions. In addition, one female had red traces on tray on
GD 28. No effect on mean maternal body weight change, mean maternal
corrected body weight change or mean maternal food consumption was
observed at this dosage. No treatment-related necropsy findings were
observed in the dams at the macroscopic examination.

At Caesarean section, no effect on litter parameters was observed. No
treatment-related findings were observed at the external, visceral or
skeletal fœtal examination.

In conclusion, a dose level of 250 mg/kg/day, was considered to cause
clear maternal toxicity but no developmental toxicity.

EVALUATION, SUMMARY AND CONCLUSION BY REGULATORY AUTHORITY

Name of authority	Pesticides Safety Directorate, UK

Reviewer’s Comments	Reliability rating: totally reliable

The study (M-273153-01-2) is fully compliant with OECD 414 (2001),
although only has a control and one treatment group, This study was
designed to supplement the first study.  Dosing from Day 6-28 of
gestation is acceptable.

	In the only treatment group there two dams were sent to necropsy on GD
28 after aborting clinical signs in one/both of these two animals
included no/few faeces, mucoid faeces, yellow sediment in the urine,
anogenital soiling, red traces on tray and placental tissue on tray.
Both females displaying marked bodyweight loss during most the dosing
period. In both cases, the bodyweight loss was associated with a marked
reduction in food consumption. There were no marked effects on
bodyweight gain in the other females.

There were no effects on litter parameters.

External Observations: At 250 mg/kg/day one fetus had multiple
malformations consisting of macroglossia, small naris, head misshapen,
malpositioned or elongated digit, and fleshy hindpaw, tail and neck. In
the control group four fetuses from separate litters had different
malformations consisting of cleft palate, absent eye bulge,
malpositioned umbilical cord and ectrodactyly of forepaws. As these
malformations occurred as isolated incidences, they were considered to
be incidental. The only anomalies observed in the treated group were
short tail in one fetus and swollen abdomen in two fetuses from separate
litters. At such low incidences these findings were considered to have
occurred by chance.

Visceral Observations.     At 250 mg/kg bw/day, one fetus had the
following malformations: ventricular septum defect in the cranial
region, left and right ventricles of the heart enlarged plus dilated
pulmonary trunk. In the control group there were four fetuses from
separate litters with malformations. One fetus had dilated cerebral
lateral ventricles, retinal fold and microphthalmia (unilateral) plus a
small thalamus, a second fetus had a ventricular septum defect in the
median region of the heart and a diaphragmatic hernia, diaphragmatic
hernia was also observed in a third fetus whilst retroesophageal aortic
arch was noted in a fourth fetus. In isolation, the malformations
observed at 250 mg/kg bw/day are considered to be incidental.

Two fetuses from separate litters at 250 mg/kg bw/day had the anomaly
misshapen kidney(s), compared with no incidences in the control group.
The mean percentage of fetuses affected was within the historical
control range, whilst the percentage of litters affected was marginally
outside the historical control range.

The incidence of the variant findings misshapen thymus and small gall
bladder, were higher at 250 mg/kg bw/day compared with the controls.
However, as the mean percentage of fetuses with misshapen thymus and
percentage of litters affected in the treated group was well within the
in-house historical control range, this finding was considered to be
unrelated to treatment.

The mean percentage of fetuses with a small gall bladder and percentage
of litters affected at 250 mg/kg bw/day were very marginally outside the
in-house historical control range. To further evaluate the relevance of
the slightly higher incidence of misshapen kidney(s), and small gall
bladder at 250 mg/kg bw/day, statistical analysis was conducted on these
parameters at both the fetal and litter level. As neither parameter was
statistically significantly increased at the fetal or litter level,
these findings were considered to have occurred by chance

EVALUATION, SUMMARY AND CONCLUSION BY REGULATORY AUTHORITY

Name of authority	Pesticides Safety Directorate, UK

	Observation: 	Dose level (mg/kg bw/day)	Historical Control Range

0	250

misshapen kidney(s)

	Mean % fetuses affected	0.0	0.9	0.0-0.9

	Mean % litters affected	0.0	8.7	0.0-8.3

	misshapen thymus

	Mean % fetuses affected	7.0	8.5	3.5-13.4

	Mean % litters affected	33.3	52.2	20.8-57.1

	small gall bladder

	Mean % fetuses affected	0.0	1.5	0.0-1.3

	Mean % litters affected	0.0	13.0	0.0-12.5

	Skeletal Observations: At 250 mg/kg bw/day, two fetuses from separate
litters had malformations; one fetus had fused thoracic centrum whilst
the second fetus had fused caudal vertebrae which were malpositioned and
bipartite. In the control group there were four malformed fetuses from
four separate litters. Two fetuses had total sternoschisis with absent
lst digit on forepaws in one case. The two other fetuses had fused
centrum in the thoracic or lumbar region, plus absent radius and lst
digit of forepaws in one case. As the malformations occurred as isolated
incidences at 250 mg/kg bw/day, they were considered to be incidental.

The occurrence of anomalies was very similar between the treated and
control groups, occurred at a higher frequency in the control group, or
as isolated findings, and were therefore regarded as chance findings.
The variant findings of 5 distal phalanges of hindpaws unossified, and
less than 15 caudal vertebrae, occurred at a higher incidence in the
treated group compared to the controls. However, as the incidence for
both mean percentage of fetuses affected and percentage of litters
affected for each parameter was within the in-house historical control
range, these findings were considered to have occurred by chance. The
following variant findings occurred at a higher incidence in the treated
group compared to the controls; l3th thoracic ribs detached, pubis
unossified, and insertion point(s) of pelvic girdle unossified. Since
the incidence of these findings was slightly outside the in-house
historical control range at the litter level or at both the fetal and
litter level, statistical analysis was conducted on these parameters at
both the fetal and litter level. None of the parameters analysed were
found to be statistically significant at either the fetal or litter
level.

Considering the skeletal observations over the two developmental studies
in the rabbit there was no dose relationships for any findings.
Therefore it is considered based on the HC data, statistical analysis
and lack of dose response these changes are non-specific or chance
findings.

	Observation: 	Dose level (mg/kg bw/day)	Historical Control Range

0	250

13th thoracic rib(s) (unilateral/bilateral) (inc. short detached

	Mean % fetuses affected	11.3	13.4	5.1-12.7

	Mean % litters affected	54.2	69.6	23.8-58.3

	hindpaws 5th middle or 5th distal phalanges unossified

	Mean % fetuses affected	0.5	1.2	0.0-1.7

	Mean % litters affected	4.2	8.7	00-13.6

	Pubis (unilateral/bilateral) unossified

	Mean % fetuses affected	0.5	1.0	0.0-1.2

	Mean % litters affected	4.2	8.7	0.0-8.3

	Insertion points (s) (unilateral/bilateral) unossified

	Mean % fetuses affected	1.0	1.3	0.0 -2.1

	Mean % litters affected	4.2	13.0	0.0-12.5

	less than 15 caudal vertebrae

	Mean % fetuses affected	0.7	2.3	0.0-2.3

	Mean % litters affected	8.3	13.0	0.0-20.8

	Conclusions	No evidence of teratogenicity was seen in this study.

No NOAEL for maternal toxicity can be determined due to the clear
maternal effects seen at the only treatment level of 250 mg/kg bw/day.
A NOEL for developmental toxicity of 250 mg/kg bw/day can be based on
the absence of any affect on fetal parameters.

HED Reviewer Comments:

Classification: Acceptable/Non-Guideline.  The study together with study
MRID No.: 47069812 satisfies the guideline requirement for a series
870.3700 developmental toxicity study in rabbits.  HED concurs with the
assignment of the NOAEL and LOAEL based on the results of the two
studies.

A.3.3	Reproductive Toxicity TC \l3 "A.3.3	Reproductive Toxicity

	870.3800 Reproduction and Fertility Effects - Rat (MRID No.: 47069815)

AE 0001789 (batch number 08466/0013, a white powder, 97.4% w/w purity)
was administered to groups of 25 male and 25 female rats at
concentrations of 0 (control), 480, 2400 and 12000 ppm in their diet
continuously throughout a multi-generation study. Test substance intake
was determined as follows based on consumption during the premating
period:

F0: 39.2, 202.3 and 1006.9 mg/kg (males); 55.3, 260.3 and 1350.2 mg/kg
(females).

F1: 50.3, 251.5 and 1292.4 mg/kg (males); 68.5, 323.7 and 1696.5 mg/kg
(females).

Test diets were fed continuously to male and female rats in the F0
parental generation throughout premating, mating, gestation, and
lactation periods. After weaning of the F1 generation at 4 weeks of age,
selected weanlings were maintained in their same dietary groups through
maturation, mating, gestation, and lactation.  The final necropsy took
place when the F2 offspring were weaned at 4 weeks of age.

Body weights, food consumption and clinical signs were monitored
regularly. Reproductive parameters such as mating performance,
fertility, duration of pregnancy, oestrus cycling and sperm analyses
were examined in F0 and F1 rats. Litter parameters such as litter size,
gender ratio, pup weight, pup viability, body weight gain and clinical
signs were studied in F1 and F2 offspring. Developmental milestones were
evaluated in F1 weanlings. At necropsy, implantation sites in F0 and F1
females were recorded. Selected organs were weighed in adult rats and
weanlings and histopathology including ovarian follicle staging (only
F1) was performed in a number of organs of F0 and F1 parental rats.

At 12 000 ppm:

Mortality was increased in F0 and F1 females due to poor general
condition and severe renal alterations. Additionally an increase in
water intake and urination was found in F1 rats. Body weight gain was
decreased in F1 males (slightly) and females during the premating phase
and in F0 and F1 females during gestation and lactation, whereas food
intake data were unchanged.

At necropsy thin female rats were found more often in treatedgroups than
at 0 ppm. Additionally, kidneys of F1 females were found to be
discoloured, enlarged and/or changed at their surface. In F0 and/or F1
females, ovaries showed an increased incidence of foamy corpus luteum
cells, the uterus exhibited more unregressed implantation sites and the
vagina showed a mucified epithelium more often. The number of primary
follicles and corpora lutea was unchanged.

In the kidneys of F0 and F1 rats, precipitates of foreign material were
found more frequently in the collecting ducts and renal pelvis. As
sequels of these precipitations, transitional cell hyperplasia, tubular
basophilia, intratubular granulocytes, papillary necrosis, medullary
fibrosis, renal pelvic dilation and/or vaculation/inclusions in the
proximal tubules were observed.  In females these findings correlated
with increased relative kidney weights. Hypertrophy was recorded in the
ureters and/or hyperplasia of the transitional epithelium of the urinary
bladder was present more frequently in females.

In the adrenal cortex the incidence of hypertrophy of the zona
glomerulosa was increased mostly in females secondary to marked renal
lesions. In the liver hepatocellular glycogen content was reduced in F0
and F1 females. In the F0 and F1 generations, increased ferrous pigment
deposits, congestion and increased haemopoiesis were noted in the spleen
and absolute and relative spleen weights were increased.

Sperm analyses and examination of œstrus cycling revealed no
substance-related effects. F0 females exhibited a reduced rearing index
due to maternal death during lactation.

There was a reduced viability index in F1 pups (including 3 total litter
losses) and a reduced lactation index in F1 and F2 pups. More pups
showing clinical findings (paleness, cold to touch, missing milk spot,
thin) were evident. The pup/litter weights were depressed at 12000 ppm.
Due to depressed body weight, vaginal opening in F1 females was delayed.
Ano-genital distance measurements in F2 pups did not indicate any
treatment-related effect.

At 2400 ppm:

Treatment-related changes were limited to histopathological changes in
the kidneys, urinary bladder and spleen of the F0 and F1 rats. Foreign
material (F0 females) and renal pelvic dilation (F1 males) were observed
in the kidneys.  Hypertrophy was recorded in the urinary bladder in F2
females.

In F0 and F1 males, increased ferrous pigment deposits, congestion
and/or increased haemopoiesis were noted in the spleen and absolute and
relative spleen weights increased in both sexes from F0 and F1
generations.

At 480 ppm:

No treatment-related findings were reported.

Thus, under the conditions described, the parental systemic LOAEL in
rats was 2400 ppm (corresponding to 202 mg/kg bw/day in males and 260
mg/kg bw/day in females), and the parental systemic NOAEL was 480 ppm
(39 mg/kg bw/day in males and 55 mg/kg bw/day in females), based on
organ changes in the spleen and urinary tract.

The NOAEL for parental reproductive parameters was 2400 ppm (173 mg/kg
bw/day) due to a reduced rearing index.

The offspring NOAEL (=NOEL) was at 2400 ppm (173 mg/kg bw/day) due to
pup weight decrease.

EVALUATION, SUMMARY AND CONCLUSION BY REGULATORY AUTHORITY

Name of authority

	Pesticides Safety Directorate, UK

Reviewer’s Comments	Reliability rating: Totally reliable

The study [M-283510-01-2] is fully compliant with OECD 416 (2001)

PSD has concerns regarding the use of the Bonferroni correction: the
Notifier is therefore requested to provide a reanalysis of the
statistical significance of the relevant parameters without the use of
this correction.

	Evaluation of vaginal smears performed at the end of the premating
periods indicated that there were no treatment related effects on the
oestrus cycle in F0 or F1 rats. Sperm parameters were investigated in
controls and top dose animals; there were no treatment related effects
on epididymal sperm count, sperm motility and morphology and testicular
spermatids counts in F1 or in F2 rats.

The parameters of the reproductive performance such as insemination,
fertility and gestation indices as well as gestation length and the
number of litters born were not influenced by treatment.

Low fertility indices were noted in 2400 and 12000 ppm F0 pairs followed
by a low number of litters and pups born, if data of the scheduled
co-housing period is considered alone. However all six 12000 ppm F0
females failing to litter after planned cohabitations were co-housed
additionally with males of the same group already shown to be fertile.
All of these females had pups (no further litter data were recorded from
these females) demonstrating that at 12000 ppm 25 of 25 F0 females were
fertile and that there was no effect of the compound on fertility.
Therefore given no indication of test substance-induced infertility was
found at other parameters such as spermatology and histology of
reproductive organs and all 12000 ppm F1 pairs had been shown to be
fertile these low fertility indices were likely incidental.

The rearing index was reduced at 12000ppm for F0 females mostly as
result of intercurrent deaths (resulting from renal lesions caused by
precipitation of foreign material in the kidneys and lower urinary
tract) of females during the lactation period.

The litter data such as mean number of implantations, prenatal loss,
live birth index, sex distribution, numbers of pups born and litter
sizes were not affected by treatment.

The viability index was unchanged up to 2400 ppm and reduced at 12000
ppm for F1 pups as a result of an increase in pup deaths including 2
total litter losses. The viability of F2 pups was unchanged up to 12000
ppm. The lactation indices were slightly reduced for F1 and F2 pups at
12000 ppm.

At 12000 ppm higher number of pups found to be pale, cold to touch, thin
and/or showing no milk spot. The development of pup litter weights were
unaffected up to 2400ppm and depressed at 12000ppm. The effects on pup
weights were not solely due to reduced food consumption.  Developmental
milestone measurements in F1 pups revealed a slight prolongation of
vaginal opening likely due to the retarded bodyweights. Anogenital
distances in F2 pups were not changed up to 12000 ppm.

At pup or weanling necropsies no remarkable microscopical findings were
noted in the F1 or F2 generations up to 12000 ppm. Changed weights of
brain, thymus and/or spleen of pups were likely to be a result of the
depressed bodyweights.

The treatment-related general toxicity findings in this study are
consistent to those seen in the 90 day study in the rat in terms of dose
level and organs/tissues affected.

.  .

	Mortality was increased at 12000 ppm in F0 and F1 females due to
clinical findings of poor general condition and severe renal effects.
Additionally in this dose an increase in water intake and urination was
noted in F1 rats.

At 12000 ppm the bodyweight gain was decreased in F1 males and females
during the premating phase and in F0 and F1 females during gestation and
lactation.

Gross necropsy revealed discolouration, enlargement and/or changed
surface in the kidneys of 12000 ppm F1 females. Microscopic examination
of the kidneys of 12000ppm rats precipitates of foreign material were
found more frequently in the collecting ducts and renal pelvis. In the
kidneys of both 2400 and 12000 ppm parental animals transitional cell
hyperplasia, tubular basophilia, intratubular granulocytes, papillary
necrosis, medullary fibrosis, renal pelvic dilation and/or
vacuolation/inclusions in the proximal tubules were observed. At 12000
ppm in females these findings correlated with increased relative kidney
weights. At doses >2400ppm hypertrophy was recorded in the ureters
and/or hyperplasia of the transitional epithelium of the urinary bladder
was present more frequently in females

In high dose F0 and/or Fl females ovaries showed an increased incidence
of foamy corpus luteum cells, the uterus exhibited more unregressed
implantation sites and the vagina showed a mucified epithelium more
often. The number of primary follicles and corpora lutea were unchanged.

In the adrenal cortex the incidence of hypertrophy of the zona
glomerulosa was increased at 12000 ppm mostly in females likely as a
secondary consequence of the marked renal lesions.

In the liver hepatocellular glycogen content was reduced in 12000 ppm F0
and F1 females.

In the spleen increased ferrous pigment deposits, congestion and
increased haemopoiesis were noted at doses >2400 ppm in males and/or
females (F0 and F1). At doses >2400 ppm absolute and relative spleen
weights increased in FO and F 1 rats.

From 2400 ppm onwards thyroid weights were increased in F1 females,
although, without any morphological correlate.

Conclusions	Dosing with cyprosulfamide did not directly cause any
reproductive toxicity a reduced rearing index due to maternal toxicity.
Therefore the NOAEL for reproduction was 2400 ppm (173 mg/kg bw/day).

A parental NOAEL of 480 ppm (equivalent to 39 mg/kg bw/day in males and
55 mg/kg bw/day in females), was based on organ changes in the spleen
and urinary tract.

The offspring NOAEL was at 2400 ppm (equivalent to 173 mg/kg bw/day) due
to pup weight decrease.

HED reviewer comments:

Classification: Acceptable/Guideline.  This study satisfies the
guideline requirement for a series 870.3700 reproduction study in rats.
HED concurs with the assignment of the NOAEL and LOAEL.

A.3.4	Chronic Toxicity TC \l3 "A.3.4	Chronic Toxicity

	870.4100a (870.4300) Chronic Toxicity – Rat

See under Carcinogenicity study - rat.

	870.4100b Chronic Toxicity - Dog (MRID No.: 47069816)

 Technical grade AE 0001789 (batch 08466/0013, 97.4 to 97.5% of purity)
was administered in the diet to Beagle dogs (4/sex/dose) at dose levels
of 0, 1000, 2500, and 8000 ppm for 370-372 days. Clinical observations
were conducted daily, food consumption was measured daily, and body
weights were taken weekly. Ophthalmic examinations were performed once
pre-exposure and just prior to necropsy. Clinical chemistry and
haematology measurements were taken once pre-exposure and during study
weeks 12, 26, 39, and 53. Urinalysis measurements were taken once
pre-exposure and during study weeks 13, 26, 39 and 53. A gross necropsy
was performed, organ weights were taken, and tissues were examined
microscopically.

At 8000 ppm

At gross pathology treatment-related findings occurred in two 8000 ppm
group males: multiple raised black zones in the mucosa of the urinary
bladder; multiple yellow pinpoint calculi in the urinary bladder; and
thickness and abnormal consistency of the urinary bladder.

Treatment-related microscopic findings were noted in the kidney and the
urinary bladder.

In the urinary bladder, stone(s) were observed at the microscopic level
in one male. In two males, slight to moderate chronic mucosal
haemorrhage and minimal to slight urothelial hyperplasia were noted.
Stone(s) were also observed at the microscopic level in one female.
Minimal suburothelial mononuclear cell infiltrates were observed in one
male and one female. In the kidney, one female had some microscopic
intrapelvic stones. Associated with these stones, there were other
minimal changes such as pelvic epithelium hyperplasia, pelvic epithelial
erosion and suburothelial mononuclear cell infiltrate.

At 2500 and 1000 ppm

No treatment-related effects were observed.

In conclusion, the NOEL was 2500 ppm (66 and 67 mg/kg/day for males and
females, respectively).

EVALUATION, SUMMARY AND CONCLUSION BY REGULATORY AUTHORITY

Name of authority	Pesticides Safety Directorate, UK

Reviewer’s Comments	Reliability rating: Totally reliable

	The study (M-283194-02-1) is fully compliant with OECD 409 (1998).
Although reticulocyte counts are listed under haematological
investigations in the study no results are reported, however red cell
morphology has been assessed

	Haematological examinations revealed statistically significant
decreases in RBC, haemoglobin, and hematocrit in males at 2500 and 8000
ppm at 6, 9, and 12 months.

 These treatment related finding were similar to the mild anaemia
observed in the 90 day dog study although in the opposite sex. However
looking at the trend over time the dose relationship is marginal and in
addition all values were well within the historical control range.
Overall the changes are not considered biologically significant.

Haematological 	Dose Group (ppm)

	Findings	males

0	1000	2500	8000

	RBCs (106/mm3)	day -4	6.83	6.75	6.33	6.63

3 months	7.27	6.72	6.36	6.54

6 months	7.53	7.22	7.02*	6.82*

9 months	7.88	7.56	7.14*	6.98*

12 months	7.69	7.65	7.6*	7.12*

	Hgb (g/dl)	day -4	16.1	15.7	14.8	15.1

3 months	16.9	15.4	15.0	15.4

6 months	17.5	16.6	16.7	16.1

9 months	18.3	17.3	16.8*	16.3*

12 months	18.5	18.0	17.6*	17.1*

	Hct (%)	day -4	45.3	44.9	42.5	44.0

3 months	47.3	43.3	42.1	43.7

6 months	48.7	46.2	46.7*	44.6*

9 months	53.5	50.6	49.5	48.1

12 months	50.6	49.7	48.6	47.4

	Haematological	FEMALES

	Findings	0	1000	2500	8000

	RBCs (106/mm3)	day -4	6.24	6.52	6.86	6.82

3 months	6.29	6.77	6.96	6.57

6 months	6.37	6.73	6.97	6.74

9 months	6.87	7.16	7.21	7.35

12 months	6.94	6.79	7.29	6.56

	Hgb (g/dl)	day -4	14.4	15.1	16.4	16.1

3 months	15.0	15.9	16.8	15.8

6 months	15.0	15.8	17.1	16.3

9 months	15.9	16.8	17.3	17.4

12 months	16.7	16.3	17.8	16.2

	Hct (%)	day -4	41.3	43.6	47.3	46.5

3 months	41.6	44.2	47.4*	45.2*

6 months	41.6	43.9	47.3	45.6

9 months	46.9	48.8	50.5	52.0

12 months	46.4	45.1	49.7	45.1

	Urinalysis revealed no clear treatment related effects.

	Conclusions	Following one years dietary treatment with cyprosulfamide
there were clear treatment related findings at the top dose only (8000
ppm) with the urinary bladder and kidney as the target organs.

In the urinary bladder gross pathology revealed multiple raised black
zones in the mucosa, multiple yellow pinpoint calculi and thick,
abnormal consistency in two high dose males. Microscopically stone(s)
were observed in one high dose male and female. In 2 males slight to
moderate chronic mucosal haemorrhage and minimal to slight urothelial
hyperplasia were noted. Minimal suburothelial mononuclear cell
infiltrate was observed in one male and one female.

In the kidney, one female had some microscopic intrapelvic stones.
Associated with these stones, there were other minimal changes: pelvic
epithelium hyperplasia, pelvic epithelial erosion and suburothelial
mononuclear cell infiltrate.

In addition cyprosulfamide appeared to cause mild anaemia with males at
2500 and 8000 ppm at 6, 9, and 12 months. However the dose relationship
is marginal and in addition all values were well within the historical
control range. Overall the changes are not considered biologically
significant.

A NOAEL of 2500 ppm (equivalent to mean achieved dietary intakes of 66
and 67 mg/kg bw/day in males and females respectively) can be
determined for this study, based on the clear effects on the kidney and
urinary bladder.

HED Reviewer Comments.

Classification:  Acceptable/Guideline.  This study satisfies the series
870.4100 guideline requirement for a chronic oral study in dogs. HED
concurs with the assignment of the NOAEL and LOAEL.

A.3.5	Carcinogenicity TC \l3 "A.3.5	Carcinogenicity

	870.4200a Carcinogenicity Study - rat (Combined chronic
feeding/carcinogenicity, MRID No.: 47069817)

AE 0001789, (batch number 08466/0013, a white powder, 97.4 to
97.5 % w/w purity) was administered to groups of 60 male and 60 female
Wistar rats by continuous dietary treatment at 1000, 4000 and
8000 ppm, corresponding to 39, 159 and 321 mg/kg/day in males and 56,
220 and 447 mg/kg/day in females, respectively, over a 24-month period.
Additionally, groups of 10 male and 10 female rats were treated at 200,
1000, 4000 and 8000 ppm AE 0001789, corresponding to 9, 45, 181 and
364 mg/kg/day in males and 13, 62, 249 and 491 mg/kg/day in females,
over a 12-month period to investigate chronic toxicity only. Mortality
and clinical signs were checked daily. Detailed physical examinations
including palpation for masses were performed at least weekly throughout
the study. Body weight was recorded weekly for the first 13 weeks, then
approximately every 4 weeks thereafter. Food consumption was recorded
twice weekly for the first 6 weeks of the study, then weekly up to
Week 13, then every 4 weeks thereafter. Ophthalmology examinations were
performed on all animals during acclimatization and after approximately
12 and 24 months. Neurotoxicity assessment (motor activity, sensory
reactivity and grip strength) was performed on all surviving animals
allocated to the chronic phase of each group during Month 12.
Haematology and clinical chemistry determinations and urinalysis were
performed during Months 4, 6/7, 12, 18 and 24/25 on selected animals. At
the scheduled chronic and carcinogenicity phase sacrifice, selected
organs were weighed and designated tissues sampled and examined
microscopically.

At 8000 ppm:

The mortality rate was higher in females than in the corresponding
control group after 24 months of treatment and was largely due to
secondary treatment-related nephropathy following product
administration.

Treatment-related clinical signs consisted of soiled anogenital region
in approximately 14 to 20% of animals of both sexes during the first
year of treatment and in 53% of males and 31% of females during the
second year of treatment. In addition, a slightly increased incidence of
red genital discharge, reduced motor activity, general pallor, wasted
appearance and hunched posture was noted in females, mainly during the
second year of treatment. Mean cumulative body weight gain was reduced
during the first week of treatment by 7 and 12% in males and females,
respectively, compared to the controls. Thereafter, mean body weight and
body weight gain parameters were comparable to controls in both sexes
throughout the study.

Urinalysis revealed the presence of sulfonamide-like crystals throughout
the study in both sexes, the effect being more pronounced in females
than in males.

At the 12-month interim sacrifice, treatment-related non-neoplastic
findings were seen microscopically in the kidney and the urinary
bladder. In the kidney, a tendency towards a higher incidence and/or
increased severity of treatment-related findings observed in the
carcinogenicity phase, i.e. unilateral pelvic dilatation, bilateral
papillary necrosis/loss/scarring, bilateral cortical/medullary scarring
and pelvic epithelium hyperplasia, was noted in one or both sexes.
Urothelial hyperplasia was noted in the urinary bladder in both sexes.
In addition, a treatment-induced nephropathy, which was considered to be
the cause of death, was observed in a prematurely sacrificed female
allocated to the chronic phase. Treatment-induced nephropathy was
characterized by stones, papillary necrosis/loss/scarring, collecting
duct hyperplasia, bilateral cortico-medullary scarring, pelvic
dilatation and/or bilateral cortex/medulla tubular dilatation.

For animals allocated to the carcinogenicity phase of the study, 14/31
premature deaths in females were attributed to a treatment-induced
nephropathy. Treatment-related macroscopic findings consisted of
abnormal shape, pelvic dilatation, irregular surface, gritty content and
stones in the kidney in females and stones in the urinary bladder in
males.

At the carcinogenicity phase (24-month) sacrifice, treatment-related
macroscopic findings in the kidney consisted of abnormal shape, pelvic
dilatation and gritty content in both sexes and irregular surface and
stones in females only. Stones were also observed in the urinary bladder
in both sexes. At the microscopic examination of unscheduled and
scheduled deaths, treatment-related effects were found in the urinary
tract, i.e. kidney, urinary bladder, and ureters. In the kidney, a
treatment-induced nephropathy was observed in both sexes, with a higher
severity in females. A slightly increased severity in simple pelvic
epithelium hyperplasia was noted in both sexes. Nodular pelvic
epithelium hyperplasia was observed in 2/60 females and a higher
incidence of intrapelvic mineral deposit was noted in females.
Intraluminal stones and multifocal/diffuse urothelial hyperplasia were
observed in the urinary bladder and in a number of ureters in both
sexes. An increased incidence of the commonly occurring lesion diffuse
bilateral tubular degeneration of the testis and bilateral oligospermia
of the epididymis was observed.

Neoplastic changes comprised a treatment-related transitional cell
carcinoma in the kidney of one male and a transitional cell carcinoma in
the urinary bladder of one female. These findings were considered to be
secondary to the combination of hyperplastic and inflammatory changes
associated with the presence of stones.

At 4000 ppm:

Treatment-related clinical signs consisted of soiled anogenital region
in approximately 5 to 9% of the animals of both sexes during the first
year of treatment and in 35% of males and 22% of females during the
second year of treatment. In males, mean cumulative body weight gain was
reduced during the first week of treatment by 5%, when compared to
controls.

Urinalysis revealed the presence of sulfonamide-like crystals throughout
the study in both sexes, the effect being more pronounced in females
than in males.

At the carcinogenicity phase (24-month) sacrifice, treatment-related
macroscopic findings consisted of pelvic dilatation, gritty content and
stones in the kidney in females and stones in the urinary bladder in
1/34 females. At the microscopic examination of unscheduled and
scheduled deaths, treatment-related effects were found in the kidney and
urinary bladder. In the kidney, stones, collecting duct hyperplasia and
bilateral cortex/medulla tubular dilatation were observed in females and
pelvic dilatation in both sexes. A slightly increased severity in simple
pelvic epithelium hyperplasia was noted in both sexes. Intraluminal
stones and multifocal/diffuse urothelial hyperplasia were observed in
the urinary bladder in females.

At 1000 ppm:

The only treatment-related changes observed at this dose level were a
slightly increased incidence of soiled anogenital region at the physical
examination in males and a tendency towards lower urinary protein levels
at urinalysis performed in Month 6/7, 12 and 18 in males. These minor
changes in the absence of corresponding histopathological findings were
considered not to be adverse.

At 200 ppm:

No treatment-related changes were observed in either sex.

In conclusion, dietary administration of AE 0001789 over a 24-month
period to the Wistar rat, at dose levels up to 8000 ppm (corresponding
to 321 mg/kg/day in males and 447 mg/kg/day in females), produced a
transitional cell carcinoma in the kidney of one male and a transitional
cell carcinoma in the urinary bladder of one female. The incidence of
these tumors was very low and was considered to be secondary to the
chronic hyperplastic changes resulting from chronic irritation due to
the presence of stones. Thus, AE 0001789 was considered not to be
directly carcinogenic in the rat.

The No Observed Adverse Effect Level (NOAEL) over a 12-month period of
dietary administration with AE 0001789 to the Wistar rat was 4000 ppm
in both sexes (equivalent to 181 mg/kg body weight/day in males and
249 mg/kg body weight/day in females).

The NOAEL over a 24-month period of dietary administration with
AE 0001789 to the Wistar rat was 1000 ppm in both sexes (equivalent to
39 mg/kg/day in males and 56 mg/kg/day in females).

EVALUATION, SUMMARY AND CONCLUSION BY REGULATORY AUTHORITY

Name of authority	Pesticides Safety Directorate, UK

Reviewer’s Comments	Reliability rating: Totally reliable

The study (M-281767-01-2) is fully compliant with OECD 453.

During the first year of treatment the mortality rate was not affected
by treatment in either sex.

After at least 104 weeks of treatment the mortality rate was not
affected by treatment in males and was slightly higher at 8000 ppm in
females compared to controls, where 14 out of 31 unscheduled mortalities
were attributed to treatment-related nephropathy. However survival rates
in males across all groups was poor with survival rates at 104 weeks
falling to 41.1%, 49.2%, 27.4%, and 34.4% at 0, 1000, 4000 and 8000 ppm
respectively.

Survival in males at doses of 4000 ppm and 8000 ppm drop below 50% at
around week 94. This weakens the carcinogenicity assessment at these
dose levels.

	Week	Survival in carcinogenicity group (60 animals/sex/group)

M	F

0	1000	4000	8000	0	500	2500	5000

	60	57	57	53	51	58	57	59	58

95.0%	95.0%	88.3%	85.0%	96.7%	95.0%	98.3%	96.7%

	75	52	51	47	45	53	51	58	49

86.7%	85.0%	78.3%	75.0%	88.3%	85.0%	96.7%	81.7%

	90	42	43	36	35	48	47	51	42

70.0%	71.7%	60.0%	58.3%	80.0%	78.3%	85.0%	70.0%

	100	35	32	21	25	42	45	42	31

58.3%	53.3%	35.0%	41.7%	70.0%	75.0%	70.0%	51.7%

	105	27	29	16	20	38	36	36	31

45.0%	48.3%	26.7%	33.3%	63.3%	60.0%	60.0%	51.7%

Unlike the previous short term studies there was no clear evidence of
effects on haematological parameters.

Sulfonamide-like crystals were observed at 8000 and 4000 ppm in both
sexes. This finding was dose-related in terms of incidence and/or
severity on most occasions, the effect being more pronounced in females
than in males. No sulfonamide-like crystals were observed at 1000 ppm,
200 ppm or in control animals in either sex.

A lower urinary protein levels was noted throughout the study in both
sexes at 8000 and 4000 ppm (except for females at Month 24) compared to
controls. The same variation was seen in males at 1000 ppm at Month 6/7,
12 and 18. This change was considered not to be adverse in view of its
low magnitude.

At Month 12, a higher urinary volume was noted at 8000 ppm in females
(+59%, p≤0.05). This change was not observed during the second year of
the study.

Gross necropsy of both premature decedents and at terminal sacrifice
revealed treatment related changes in the kidneys and urinary bladder at
doses >4000 ppm. Findings in the kidneys included abnormal shape, pelvic
dilatation, irregular surface, gritty content and stones. In the urinary
bladder stones were observed.

Microscopic examinations at 12 month sacrifice revealed a number of
non-neoplastic changes at the top dose; a slightly higher incidence
and/or a slight increase in severity of unilateral pelvic dilatation,
bilateral papillary necrosis/loss/scarring, bilateral cortical/medullary
scarring and pelvic epithelium hyperplasia, was noted in the kidney in
one or both sexes. Treatment-related urothelial hyperplasia was noted in
the urinary bladder in a few animals of both sexes. There was no
evidence of a neoplastic effect at 12 months.

At 24 months treatment-related non-neoplastic effects were observed in
the urinary tract, i.e. kidney, urinary bladder and ureter, including a
combination of hyperplastic and inflammatory changes associated with the
presence of stones.

Treatment-induced nephropathy was observed at a higher incidence in both
sexes in the kidney at 8000 ppm, with an increased severity in females,
and at 4000 ppm in a higher incidence in females only. Treatment-induced
nephropathy is a general term which incorporates some or all of the
following findings: stones, papillary necrosis/loss/scarring, collecting
duct hyperplasia, bilateral cortico-medullary scarring, pelvic
dilatation and bilateral cortex/medulla tubular dilatation. In addition,
an increased incidence of pelvic dilatation was noted at 4000 ppm in
males.

A slight increase in severity of simple pelvic epithelium hyperplasia
was observed in both sexes at 8000 and 4000 ppm. Nodular pelvic
epithelium hyperplasia was observed in 2/60 females and a higher
incidence of intrapelvic mineral deposit was observed in 14/60 females
at 8000 ppm.

Intraluminal stones and multifocal/diffuse urothelial hyperplasia were
observed in the urinary bladder in both sexes at 8000 ppm and in females
at 4000 ppm. These lesions were considered to be treatment-related.
Suburothelial mononuclear cell infiltrate and mixed cell infiltrate were
considered to be associated lesions

There was a treatment related increase (ca three times the numbers in
controls) in the incidence of the diffuse bilateral tubular degeneration
of the testis (associated with diffuse interstitial oedema and/or
focal/multifocal mineralization in most cases) and bilateral
oligospermia of the epididymis (associated with epithelial degenerative
change) observed at 8000 ppm.

EVALUATION, SUMMARY AND CONCLUSION BY REGULATORY AUTHORITY

Name of authority	Pesticides Safety Directorate, UK

Reviewer’s Comments	Reliability rating: Totally reliable

The study (M-281767-01-2) is fully compliant with OECD 453.

During the first year of treatment the mortality rate was not affected
by treatment in either sex.

After at least 104 weeks of treatment the mortality rate was not
affected by treatment in males and was slightly higher at 8000 ppm in
females compared to controls, where 14 out of 31 unscheduled mortalities
were attributed to treatment-related nephropathy. However survival rates
in males across all groups was poor with survival rates at 104 weeks
falling to 41.1%, 49.2%, 27.4%, and 34.4% at 0, 1000, 4000 and 8000 ppm
respectively.

Survival in males at doses of 4000 ppm and 8000 ppm drop below 50% at
around week 94. This weakens the carcinogenicity assessment at these
dose levels.

	Week	Survival in carcinogenicity group (60 animals/sex/group)

M	F

0	1000	4000	8000	0	500	2500	5000

	60	57	57	53	51	58	57	59	58

95.0%	95.0%	88.3%	85.0%	96.7%	95.0%	98.3%	96.7%

	75	52	51	47	45	53	51	58	49

86.7%	85.0%	78.3%	75.0%	88.3%	85.0%	96.7%	81.7%

	90	42	43	36	35	48	47	51	42

70.0%	71.7%	60.0%	58.3%	80.0%	78.3%	85.0%	70.0%

	100	35	32	21	25	42	45	42	31

58.3%	53.3%	35.0%	41.7%	70.0%	75.0%	70.0%	51.7%

	105	27	29	16	20	38	36	36	31

45.0%	48.3%	26.7%	33.3%	63.3%	60.0%	60.0%	51.7%

Unlike the previous short term studies there was no clear evidence of
effects on haematological parameters.

Sulfonamide-like crystals were observed at 8000 and 4000 ppm in both
sexes. This finding was dose-related in terms of incidence and/or
severity on most occasions, the effect being more pronounced in females
than in males. No sulfonamide-like crystals were observed at 1000 ppm,
200 ppm or in control animals in either sex.

A lower urinary protein levels was noted throughout the study in both
sexes at 8000 and 4000 ppm (except for females at Month 24) compared to
controls. The same variation was seen in males at 1000 ppm at Month 6/7,
12 and 18. This change was considered not to be adverse in view of its
low magnitude.

At Month 12, a higher urinary volume was noted at 8000 ppm in females
(+59%, p≤0.05). This change was not observed during the second year of
the study.

Gross necropsy of both premature decedents and at terminal sacrifice
revealed treatment related changes in the kidneys and urinary bladder at
doses >4000 ppm. Findings in the kidneys included abnormal shape, pelvic
dilatation, irregular surface, gritty content and stones. In the urinary
bladder stones were observed.

Microscopic examinations at 12 month sacrifice revealed a number of
non-neoplastic changes at the top dose; a slightly higher incidence
and/or a slight increase in severity of unilateral pelvic dilatation,
bilateral papillary necrosis/loss/scarring, bilateral cortical/medullary
scarring and pelvic epithelium hyperplasia, was noted in the kidney in
one or both sexes. Treatment-related urothelial hyperplasia was noted in
the urinary bladder in a few animals of both sexes. There was no
evidence of a neoplastic effect at 12 months.

At 24 months treatment-related non-neoplastic effects were observed in
the urinary tract, i.e. kidney, urinary bladder and ureter, including a
combination of hyperplastic and inflammatory changes associated with the
presence of stones.

Treatment-induced nephropathy was observed at a higher incidence in both
sexes in the kidney at 8000 ppm, with an increased severity in females,
and at 4000 ppm in a higher incidence in females only. Treatment-induced
nephropathy is a general term which incorporates some or all of the
following findings: stones, papillary necrosis/loss/scarring, collecting
duct hyperplasia, bilateral cortico-medullary scarring, pelvic
dilatation and bilateral cortex/medulla tubular dilatation. In addition,
an increased incidence of pelvic dilatation was noted at 4000 ppm in
males.

A slight increase in severity of simple pelvic epithelium hyperplasia
was observed in both sexes at 8000 and 4000 ppm. Nodular pelvic
epithelium hyperplasia was observed in 2/60 females and a higher
incidence of intrapelvic mineral deposit was observed in 14/60 females
at 8000 ppm.

Intraluminal stones and multifocal/diffuse urothelial hyperplasia were
observed in the urinary bladder in both sexes at 8000 ppm and in females
at 4000 ppm. These lesions were considered to be treatment-related.
Suburothelial mononuclear cell infiltrate and mixed cell infiltrate were
considered to be associated lesions

There was a treatment related increase (ca three times the numbers in
controls) in the incidence of the diffuse bilateral tubular degeneration
of the testis (associated with diffuse interstitial oedema and/or
focal/multifocal mineralization in most cases) and bilateral
oligospermia of the epididymis (associated with epithelial degenerative
change) observed at 8000 ppm.

HED Comments:

Classification: Acceptable/Guideline.  This study satisfies the
guideline requirement for a series 870.4300 combined chronic feeding
carcinogenicity study in rats.  HED concurs with the assignment of the
NOAEL and LOAEL.  The HED CARC determined that the study was adequate
for assessing carcinogenic potential in the rat.

	870.4200b Carcinogenicity (feeding) - Mouse (MRID No.: 47069818)

The objective of this study was to investigate the oncogenic potential
of AE 0001789 (batch number 08466/0013, a white powder, 97.4-97.5% w/w
purity) in the C57BL/6J mouse following continuous dietary treatment at
350, 2000 and 3500 ppm for 18 months. In addition, an interim sacrifice
was performed after 52 weeks of treatment to assess chronic toxicity.

Groups of 60 male and 60 female C57BL/6J mice were initially intended to
be fed diet containing 0, 350, 2000 or 7000 ppm of AE 0001789 for at
least 78 weeks. Since the Maximum Tolerated Dose was exceeded in females
at 7000 ppm during the first weeks of treatment, an additional treatment
group (60 animals/sex) at 3500 ppm was added to the study approximately
one month after the start of treatment. The high rate of mortality
observed in both sexes at 7000 ppm led to the early termination of this
treatment group, with all surviving females being sacrificed during Week
6 (study Day 40) and all males being sacrificed during Week 46 (study
Day 320). After 52 weeks, 10 males and 10 females from each group
allocated to the chronic phase of the study were necropsied at the
scheduled interim sacrifice. The remaining 50 animals/sex/group,
allocated to the carcinogenicity phase of the study, continued treatment
until the scheduled final sacrifice of the study after at least 78 weeks
of treatment (corresponding to a mean intake of AE 0001789 over 18
months of 0, 50, 287 and 506 mg/kg/day in males and 0, 63, 354, 616
mg/kg/day in females, at 0, 350, 2000 and 3500 ppm, respectively).
Mortality and clinical signs were checked daily. Additionally, detailed
physical examinations including palpation for masses were performed
weekly throughout treatment. Body weight and food consumption were
measured weekly for the first 13 weeks of the study, then approximately
monthly thereafter. Haematology determinations were performed at
approximately 12 and 18 months from designated animals. Where possible,
blood smears were prepared from moribund animals just before sacrifice.
All animals were subjected to necropsy, with selected organs weighed at
scheduled interim and final sacrifice. Designated tissues were fixed and
examined microscopically. In addition, urinary bladder stones sampled
from one male and one female at 3500 ppm and from one male at 2000 ppm
were analyzed for the presence of test substance. For the dose group
treated at 3500 ppm, all parameters were assessed on the same study days
as the other dose groups, resulting in staggered calendar dates.

At 7000 ppm:

The dietary level of 7000 ppm exceeded the Maximum Tolerated Dose in
both males and females (marked individual body weight loss and mortality
in both sexes), leading to early sacrifice of all males and females on
Day 320 and 40, respectively. Fifteen females were either found dead or
were killed for humane reasons between Days 11 and 35. In males, 5
animals died prematurely during the first three months of treatment and
26 animals were either found dead or were killed for humane reasons
between Days 218 and 316. In both sexes, most animals showed a severe
body weight loss on the days preceding death, they had a wasted
appearance and exhibited clinical signs related to a poor health
condition on one or more occasions prior to death.

At 3500 ppm:

The mortality incidence in males was higher than in the control group
after 12 and 18 months of treatment (20.0% and 58.1%, respectively,
compared to 5.0% and 24.4% in the respective control group). The higher
mortality rate was largely due to the presence of stones within the
urinary tract, causing acute or chronic renal failure or due to
secondary treatment-related nephropathy following product
administration. Treatment-related clinical signs were confined to males
and consisted of clinical observations which were, on most occasions,
recorded on the days preceding the death or early sacrifice of the
animals and reflecting their general poor health condition. During the
first week of treatment, mean body weight of males was reduced by 4%,
whilst mean body weight gain was reduced by 33%, compared to the
controls. Thereafter, starting from Day 92, mean body weight was
slightly reduced, compared to controls, at most time points, resulting
in a reduction of 5% at the end of the treatment period (Day 540). An
overall mean cumulative body weight gain reduction of 12% was observed
by Day 540, when compared to controls. In females, mean cumulative body
weight gain was reduced by between 29 and 71% at most time points during
the last 6 months of treatment (week period 55-77), resulting in a mean
body weight reduction of 3% on Day 540, compared to controls. During the
last 6 months of treatment, mean food consumption was reduced by between
2 and 10% in females, when compared to controls.

At the 18-month haematology determination, a tendency towards lower red
blood cell count was noted in both sexes. This change was associated
with slightly lower haemoglobin concentration, haematocrit and mean
corpuscular haemoglobin concentration and slightly higher mean
corpuscular volume.

At the 12-month interim sacrifice, macroscopic findings consisted of
thickening of the mucosa and stones in the urinary bladder of 4/9 and
1/9 males, respectively. Pale kidneys and gritty content were seen in
the urinary bladder in a prematurely sacrificed male allocated to the
chronic phase. At the microscopic examination, a higher incidence of
renal cortical basophilic tubules was noted in the kidney of males. In
the urinary bladder, urothelial hyperplasia, interstitial oedema,
induced arteritis/periarteritis, suburothelial mixed cell infiltrate and
intramuscular inflammatory cell infiltrates were observed in one male.

For males allocated to the carcinogenicity phase of the study, 21/30
premature deaths were attributed to the presence of urinary stones or to
treatment-related nephropathy. In females, 2/11 deaths was attributed to
the presence of urinary stones or to treatment-related nephropathy.
Treatment-related macroscopic findings at unscheduled sacrifice
consisted of stones found in the urinary bladder of both sexes,
thickening of the mucosa in the urinary bladder and dilatation of
ureters in males. A higher incidence of enlarged kidney, renal pelvic
dilatation, pale kidney or kidney with irregular surface was observed in
males and/or females, compared to controls. The incidence of gritty
content in the kidney was increased in males and stones were observed in
the kidney of one male and one female. An increased incidence of dark
liver was noted in males. This finding was often associated with
hepatocellular hypertrophy at the microscopic examination.

At the carcinogenicity phase (18-month) sacrifice, mean terminal body
weight of males and females was 5% lower than the controls. At the
macroscopic examination, kidneys with an irregular surface were observed
in 6/20 males, compared to 1/37 males in the control group, and stones
were found in the kidney of 1/20 males, compared to no cases in the
control group. At the microscopic examination, treatment-related effects
were found in the liver and the urinary tract including kidney, urinary
bladder, prostatic urethra and ureters. In the liver, minimal to slight
centrilobular to panlobular hepatocellular hypertrophy and an increased
incidence of hepatocellular single cell necrosis were observed in males.

In the urinary bladder, stones were noted in both sexes,
hyperplastic/metaplastic changes and secondary changes due to
stones-induced irritation or urinary blockage (muscular
degeneration/hemorrhage(s) and interstitial oedema, inflammatory cell
infiltrate, mixed cell infiltrate and arteritis/periarteritis) were
noted in males and/or females. In the kidney, stones were observed in
the pelvis in both sexes and in the collecting ducts in males only, and
a higher incidence of concretions was found in the collecting ducts in
males. Induced hyperplastic changes (collecting duct and
papillary/pelvic epithelium hyperplasia) and changes resulting from
chronic inflammation or urinary blockage (atrophy/fibrosis/scar,
perirenal inflammation, glomerular chamber and cortical or
cortex/medulla tubular dilatation, parenchymal cysts and pelvic
dilatation) were observed mostly in males. In addition in males alone, a
higher incidence and severity of bilateral cortical basophilic tubules
was noted and a higher incidence of cortical and medullar tubular
mineralization was found. The incidence of cellular casts was also
increased. Stones were found in the prostatic urethra and ureter.
Urothelial hyperplasia and interstitial mixed cell infiltrate were also
observed.

All the effects seen in the urinary tract were considered to be due to a
single and chronic irritative mechanism, resulting from the formation of
stones or crystals. In addition, indirect treatment-related findings
were observed in the lymphoid system in males (apoptotic bodies in the
spleen and mesenteric lymph nodes, and lymphoid atrophy in the spleen)
and were considered to be secondary to the stress induced by the stone
deposition and/or the treatment-related nephropathy. In the heart, a
higher incidence of epicardial mixed cell infiltrate was noted in 5/50
males, when compared to no cases in controls. This minimal change only
observed in males was considered to be an indirect and non-adverse
response to the treatment.

Neoplastic changes comprised transitional cell papilloma in the urinary
bladder of 2/49 females. This finding was considered to be secondary to
the chronic hyperplastic changes due to the presence of calculi.
Analysis of urinary bladder stones taken from one male and one female at
terminal sacrifice revealed that the composition of the stones consisted
of approximately 90 to 95% of test material.

At 2000 ppm:

There was no treatment-related change in females. Mortality rate, body
weight or food consumption parameters were not affected in males. At the
12-month interim sacrifice there were no treatment-related organ weight
changes and no macroscopic or microscopic findings.

For animals allocated to the carcinogenicity phase of the study, 1/13
premature deaths in males was attributed to the presence of urinary
stones. Treatment-related macroscopic findings at unscheduled sacrifice
consisted of gritty content found in the urinary bladder of 1/13 males
and in the kidney of 2/13 males, compared to no cases in the control
group.

At the carcinogenicity phase (18-month) sacrifice, treatment-related
findings were confined to males. Stones were observed in the urinary
bladder of 1/37 males and gritty content was found in the kidney of 1/37
males at the macroscopic examination, compared to no cases in the
control group. At the microscopic examination, treatment-related effects
were found in the liver, kidney, urinary bladder, prostatic urethra and
ureters. In the liver, minimal centrilobular to panlobular
hepatocellular hypertrophy was observed in a few males. In the urinary
bladder, stones were noted in 2/49 males. Simple hyperplasia and
interstitial oedema were observed in 1/49 males and intraluminal mixed
cell infiltrate was observed in another male. In the kidney, stones were
noted in the pelvis and collecting ducts in males. Hyperplasia was