### 1 OFFICE OF CHEMICAL SAFETY AND POLLUTION PREVENTION 

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

August 18, 2015

PC Code: 128819
DP Barcode: 428390

MEMORANDUM

SUBJECT: Forchlorfenuron: Response to Comments on the Preliminary Work Plan for Registration Review: Initial Docket Case Number 7057

FROM: Christina Wendel, Biologist
Environmental Risk Branch 2 Environmental Fate and Effects Division (7507P)

THROUGH: Elyssa Arnold, Risk Assessment Process Leader Brian Anderson, Branch Chief
Environmental Risk Branch 2 Environmental Fate and Effects Division (7507P)

TO: Wilhelmena Livingston, Chemical Review Manager Cathryn Britton, Team Leader Kevin Costello, Branch Chief Risk Management and Implementation Branch 4
Pesticide Re-evaluati on Division (7508P)

The Environmental Fate and Effects Division (EFE D) has completed its response to public comments on the Preliminary Work Plan (PWP) for the Registration Review of Forchlorfenuron (Docket ID No.: EPA-HQ-OPP -2014-0641). Comments were received from KIM-C1, LLC (registrant for forchlorfenuron products), and the Center for Biological Diversity (CBD). The comments from CBD will be addressed by the Pesticide Re-evaluation Division (PRD). The comments received by KIM-C1, LLC that were relevant to EFED are summarized below, followed by EFED’s responses.

KIM-C1, LLC

Comment 1: “Under Table 1 (“Anticipated Data Needs for the Forchlorfenuron Registration Review”), page 4, “Terrestrial plant toxicity, vegetative vigor” is listed. KIM -C1, LLC has initiated a new study to replace the prior study which was classified “Supplemental”. The plant culture phase of the new study was scheduled to begin June 15, 2015. Applications will 2015.08.18 08:27:59 -04'00'

### 2 begin approximately July 1, 2015 and terminate approx. July 30, 2015 when shoot dry weight data is collected. A draft report is targeted for October, 2015 and a final report for December,
2015. ”

EFED Response: EFED is looking forward to reviewing the terrestrial plant, vegetative vigor data when it is submitted to the Agency. EFED recommends that care is taken to ensure that the study is conducted at the maxim um application rate of 0.022 lbs a.i./A. This was one of the deviations that resulted in the downgrading of the currently available vegetative vigor toxicity study, and there is uncertainty of risk at the maximum application rate. In addition, a no-effect concentration level needs to be established; this is especially vital for endangered species risk assessments.

Comment 2: The registrant does not agree with the initial conclusions of the avian reproduction study by EFED that wer e presented in the “ EFED Registration Review Problem Formulation for Forchlorfenuron Christina Wendel and Greg Orrick, PC Code: 128819, DP Barcode: 422524, January 28, 2015.” KIM -C1, LLC cite data presented in the study report (Frey LT, Martin KH, Beavers JB, Jaber M. 2014. “Forchlorfenuron: A Reproduction Study with the Northern Bobwhite”, MRID No. 49350901). Including the reproduction of Table 6, and conclusion section of the report.

“ KIM-C1, LLC respectfully requests EFED to re-examine this issue prior to completing the Final Work Plan, and any final conclusions therein, relative to the submitted avian reproduction study. ”

EFED Response: EFED has completed the final review of the avian reproduction DER for bobwhite quail (MRID 49350901). The DER can be found attached to this
memorandum in Appendix A

. Please see EFED’s response to the avian reproduction data waiver request (USEPA, 2015; DP 417353) for additional discussion regarding this study.

References:

Frey L.T., Martin K.H., Beavers J.B., and Jaber M. (2014). Forchlorfenuron: A Reproduction Study with the Northern Bobwhite. Unpublished study performed by Wildlife International, Ltd., Easton, Maryland. Laboratory report number 248-108. Study sponsored by Degussa AG, Product Line Agriculture, D-83308 Trostberg, Germany. Study completed on December 3, 2003 and report amended on March 21, 2014.
MRID 49350901.

United States Environmental Protection Agency (USEPA). 2015. Wendel, C., G. Orrick, E.
Arnold and B. Anderson. Memorandum “ Forchlorfenuron: Response to Data Waiver Requests for a Terrestrial Field Dissipat ion Study and an Avian Reproduction Study AND Summary of the Review of an Ecotoxicit y Study on the Ecological Effects of Forchlorfenuron ” DP barcode: 316642, 417353, and 420428. Dated August 11, 2015. Office of Pesticide Programs, Environmental Fate and Effects Division. Memorandum to the Registration Division.

### Data Evaluation Report on the Reproductive Effects of Forchlorfenuron on Avian Species Colinus virginianus PMRA Submission Number {......} EPA MRID Number 49350901



### Page 1 of 15 Data Requirement: PMRA Data Code {............}
EPA DP Barcode 420428 OECD Data Point {............}
EPA MRID 49350901 EPA Guideline OCSPP (formerly OPPTS) 850.2300

Test material: Forchlorfenuron Purity: 96.95% Common name: Forchlorfenuron Chemical name: IUPAC: Not Reported CAS name: N-(2-chloro-4-pyridinyl)- N’ -phenylurea
CAS No.: 68157-60-8 Synonyms: None Reported

Primary Reviewer: John Marton, Ph.D. Signature: Environmental Scientist, CDM Smith Date: 09/13/2014

Secondary Reviewer: Teri S. Myers, Ph.D. Signature: Environmental Scientist, CDM Smith Date: 09/22/2014

Primary Reviewer: N.E. Federoff Signature: Wildlife Biologist/USEPA/OPP/EFED/ERB2 Date: 08/5/2015

EPA PC Code 128819

CITATION: Frey L.T., Martin K.H., Beavers J.B., and Jaber M. 2014. Forchlorfenuron: A Reproduction Study with the Northern Bobwhite. Unpublished study performed by Wildlife International, Ltd., Easton, Maryland. Laboratory report number 248-108. Study sponsored by Degussa AG, Product Line Agriculture, D-83308 Trostberg, Germany. Study completed on December 3, 2003 and report amended on March 21, 2014. 

## Appendix A. Forchlorfenuron Avian Reproduction DER; MRID 49350901

### Data Evaluation Report on the Reproductive Effects of Forchlorfenuron on Avian Species Colinus virginianus PMRA Submission Number {......} EPA MRID Number 49350901



### Page 2 of 15 EXECUTIVE SUMMARY

The one-generation reproductive toxicity of forchlorfenuron to groups of 16 pairs of 20-week-old Northern bobwhite quail ( Colinus virginianus ) were assessed over 20 weeks. Forchlorfenuron wa s administered to the birds in the diet at nominal concentrations of 0 (negative control), 160, 400 , and 1000 mg a.i./kg diet (ppm). Mean-measured concentrations in the diet were 170, 435, and 1070 mg a.i. /kg diet, respectively. The NOAEC was determined to be <170 mg a.i./kg diet based on significant, slig htly (7.0-9.1%) dose-dependent reductions ( p < 0.05) in 14-d survivor weight at the 170, 435, and 1070 mg a.i./kg diet lev els when compared to the negative control.

There were no treatment-related mortalities, abnormal signs of toxicity, or remarkable pathological findings in any treatment group relative to the negative control. Further, mean food consumption and sex-specific adult body weight gain were comparable between the control and treatment grou ps. With the exception of 14-d survivor weight, there were no other treatment-related effects on any reproductive parameter analyzed.

This study is classified as supplemental

.

Results Synopsis

Test Organism Size/Avg. (mean Weight): 20 weeks

NOAEC: <170 mg a.i./kg diet LOAEC: 170 mg a.i./kg diet

Endpoint(s) Affected: 14-d survivor weight

### Data Evaluation Report on the Reproductive Effects of Forchlorfenuron on Avian Species Colinus virginianus PMRA Submission Number {......} EPA MRID Number 49350901



### Page 3 of 15 I. MATERIALS AND METHODS

GUIDELINE FOLLOWED: This study was conducted following guidelines outlined in the Environmental Protection Agency’s Registration Guidelines Pesticide Assessment Guidelines, FIFRA Subdivis ion E, Hazard Evaluation: Wildlife and Aquatic Organisms, Subsection 71-4; U.S. Environmental Protection Agency Series 850- Ecological Effects Test Guideline OCSPP (formerly OPPTS) Number 850.2300; OECD Guideline 206; and the ASTM “Standard Practice for Conducting Reproductive Studies with Avian Species”. There were no deviations from OCSPP 850.2300 (June 2012)
noted.

COMPLIANCE: Signed and dated No Data Confidentiality, GLP, and Quality Assurance statements were provided. This study was conducted in compliance with Good Laboratory Practice Standards as published by the U.S. Environmental Protection Agency, 40 CFR Part 160, 17 August 1989; OECD Principles of Good Laboratory Practice (ENV/MC/CHEM(98) 17); and Japan MAFF, 11 Nohsan Notification No. 6283. Agricultural Production Bureau, 1 October 1999, with the following exceptions: the test substance was not characterized in accordance with GLP standards; and, stability under storage conditions at the test site was not determined in accordance with GLP standards.

A. MATERIALS:

1. Test Material Forchlorfenuron

Description: Powder

Lot No./Batch No. : 97030010 (Batch No.)

Purity: 96.95%

Stability of compound under test conditions: Mean-measured concentrations were 170, 435, and 1070 mg a.i./kg diet, yielding recoveries of 106, 109, and 107% of nominal.

(OECD recommends water solu bility, stability in water and light, pKa, Pow, and vapor pressure of test compound)

Storage conditions of test chemicals: Stored under ambient conditions.

Physicochemical properties of Forchlorfenuron.

Parameter Values Comments

Water solubility at 20°C Not Reported
Vapor pressure Not Reported
UV absorption Not Reported
pKa Not Reported

Kow Not Reported

### Data Evaluation Report on the Reproductive Effects of Forchlorfenuron on Avian Species Colinus virginianus PMRA Submission Number {......} EPA MRID Number 49350901



### Page 4 of 15 2. Test organism :

Table 1: Test organism.

Parameter Details Remarks

Criteria

Species (common and scientific names): Northern Bobwhite quail, ( Colinus virginianus ) Recommended species include a wild waterfowl species, preferably the mallard (Anas platyrhynchos) or an upland game species, preferably the northern bobwhite (Colinus virginianus)

Age at Study Initiation: 20 weeks From the same hatch, approaching first breeding season, and have not been used in any previous testing.

Birds approaching their first breeding season should be used.

Body Weight: (mean and range) 170-216 g Weight at test initiation

Body weights should be recorded at test initiation and at biweekly intervals up to week eight or up to the onset of egg laying and at
termination.

Source: Trace Pheasantry, 288 Levengood Rd. Douglassville, PA , USA All birds should be from the same

source.

B. STUDY DESIGN:

1. Experimental Conditions

a. Range-finding study: No rang e-finding data were reported.

b. Definitive Study

### Data Evaluation Report on the Reproductive Effects of Forchlorfenuron on Avian Species Colinus virginianus PMRA Submission Number {......} EPA MRID Number 49350901



### Page 5 of 15 Table 2: Experimental Parameters.

Parameter Details Remarks

Criteria

Acclimation period:

Conditions (same as test or not):

Feeding:

Health (any mortality observed): 4 weeks

Same as test

Diet was provided ad libitum and was formulated by Agway Inc., and contained at least 27% protein, 2.5% fat, and no more than 5% fiber.

Appeared normal and healthy prior to testing

. Phenotypically indistinguishable from wild type at start of
acclimation. Basal diet contained approx. 1.1% calcium; however,
additional limestone is needed for breeding birds. Therefore, basal diets contained supplemental limestone of 5% (w/w); (38.5% Ca)
for adult birds.

Recommended observation period
includes a 2

- 3 week health observation period prior to selection of birds for treatment. Generally, birds should be healthy without excess mortality. Feeding should be ad libitum, and sickness, injuries or mortality should be
noted

.

Test duration

pre -laying exposure:

egg -laying exposure:

withdrawal period, if used: 10 weeks (7 weeks pre

- photostimulation

+ 3 weeks post

- photostimulation)

10 weeks

N/A Pre-laying exposure :
pre -photostimulation - 7 weeks; preegg laying: (w / photostimulation)

– 3 weeks (7+3 = 10 weeks). Egg-laying: Approx. 10 weeks

Recommended pre-laying exposure duration: At least 10 weeks prior to the onset of

egg -laying. Recommended exposure duration with

egg -laying:
At least 10 weeks. Recommended withdrawal period : If reduced reproduction is evident, a withdrawal period of up to 3 weeks should be added to the test phase.

### Data Evaluation Report on the Reproductive Effects of Forchlorfenuron on Avian Species Colinus virginianus PMRA Submission Number {......} EPA MRID Number 49350901



### Page 6 of 15 Parameter Details Remarks

Criteria

Pen (parental and offspring)
size:

construction materials:

number: A dult Pens: 27 x 51 cm; sloping
floors that resulted in ceiling height ranging from 20 to 25 cm.

Offspring Pens: Hatchlings housed in batteries of brooding pens 72 x 90 x 23 cm high

.

Adult Pens: Pens were constructed of galvanized wire mesh and galvanized sheeting.

Offspring Pens: External walls and ceilings were constructed of galvanized wire mesh and galvanized sheeting; floors were of galvanized wire mesh.

Adult Pens: 16 pens per treatment
with one male and one female in each pen

.

Offspring Pens: Adult Pens: GQFM Model No. 0330

Offspring Pens: Beacon Steel Co. Model B735Q

Pens Pens should have adequate room and be arranged to prevent cross
contamination. Materials
Recommended materials include nontoxic material and nonbinding material, such as galvanized steel.
Number At least 5 replicate pens should be used for mallards housed in groups of 7. For other arrangements, at least 12 pens should be used, but considerably more may be used if birds are kept in pairs. Chicks should be housed according to parental grouping.

Number of birds per pen (male:female) One male and one female Sex of birds were determined by a visual examination of plumage.

One male and one female per pen should be used. For quail, one male and two females should be used. For ducks, two males and five females
should be used.

Number of pens per group/treatment negative control:
solvent control:
treated: 16 N/A
16 At least 12-16 pens should be used, but considerably more if birds are kept in pairs.

Test concentrations (mg a.i./kg diet)
nominal:

measured: 0 (negative control), 160, 400, and 1000 mg a .i./kg diet

0, 170, 435, and 1070 mg a.i. /kg
diet (mean-measured) Recommended test concentrations
include at least two concentrations other than the control; three or more will provide a better statistical analysis. The highest test concentrations should show a significant effect or be at or above the actual or expected field
residue level.

### Data Evaluation Report on the Reproductive Effects of Forchlorfenuron on Avian Species Colinus virginianus PMRA Submission Number {......} EPA MRID Number 49350901



### Page 7 of 15 Parameter Details Remarks

Criteria

Maximum labeled field residue anticipated and source of information: Not reported

The highest test concentrations should show a significant effect or be at or above the actual or expected field residue level. The source (i.e.,
maximum label rate in lb a .i./A and ppm), label registration no., label date,
and site should be cited ]

Solvent/vehicle, if used type:

amount: N/A; a solvent was not used
Recommended solvents include corn oil or other appropriate vehicle not more than 2% of diet by weight

Was detailed description and nutrient analysis of the basal diet provided (Yes/No) Yes A commercial breeder feed or an equivalent that is appropriate for the test species is recommended.

Preparation of test diet Test diets were prepared by mixing forchlorfenuron into a premix that was used for weekly preparation of the diet. Control and treatment diets were prepared weekly and were adjusted for the purity of the test material. A premixed diet containing the test substance should be mechanically mixed with basal diet. If an evaporative vehicle is used, it should be completely evaporated prior to feeding.

Indicate whether stability and homogeneity of test material in diet determined (Yes/No) Yes

Were concentrations in diet verified by chemical analysis? Yes LOD=1.0 μ g a.i./mL = 50.0 ppm a.i LOQ = 75.0 ppm a.i.

Did chemical analysis confirm that diet was stable and homogeneous? Yes

Feeding and husbandry Feeding and husbandry conditions appeared to be adequate, given guideline recommendations. Basal diets contained supplemental
limestone 5% (w/w); (38.5% Ca)
for adult birds . For offspring the
basal diet did not contain either the
test substance or the additional supplemental limestone.

### Data Evaluation Report on the Reproductive Effects of Forchlorfenuron on Avian Species Colinus virginianus PMRA Submission Number {......} EPA MRID Number 49350901



### Page 8 of 15 Parameter Details Remarks

Criteria

Test conditions (pre-laying) temperature:

relative humidity:

photoperiod: 20.6 ± 1.3°C (SD)

31 ± 11% (SD)

8L: 8D for acclimation and the
first seven weeks of the test; then
it increased to 17L: 7D (at beginning of week 8 to induce egg laying

– maintained at that length until adult birds were
euthanized ) Throughout the test, birds received a mean of approx. 217 lux (~20 foot candles) of illumination by fluorescent lights

– closely approximated noon

- day sunlight. The air handling system in study

room

– designed to vent up to 15 room air volumes every hour and replace them with fresh air.

Recommended temperature:
about 21°C (70°F) Recommended relative humidity:
about 55% Recommended lighting First 8 weeks: 7 h per day. Thereafter: 16 -17 h per day. At least 6 foot -candles are
recommended at bird level.

Egg Collection and Incubation

Egg collection and storage:
collection interval:

storage temperature:

storage humidity:

Incubation :

temperature:

humidity: Daily

14.0 ± 0.1°C (SD)

– mean temp.

81 ± 6% (SD)

– mean relative humidity

37.0 ± 0.0°C (SD)

– avg. temp.

54 ± 0.4% (SD)

– avg. relative humidity Collected daily from all pens, when available. Eggs were stored in a cold room until incubation. All eggs laid in a weekly interval were
considered as one lot.

NatureForm Incubator (Model No.
NMC -4000 or NMC-7000), equipped with pulsator fan and
blades

– produced mild breathing

movement ; also equipped w/ automatic egg rotation device designed to rotate eggs 90 q every hour through 21 days of incubation.

Eggs should be collected daily; recommended egg storage temperature is approximately 16 °C (61°F); recommended humidity is approximately 65%. Recommended collection interval: daily

### Data Evaluation Report on the Reproductive Effects of Forchlorfenuron on Avian Species Colinus virginianus PMRA Submission Number {......} EPA MRID Number 49350901



### Page 9 of 15 Parameter Details Remarks

Criteria

Were eggs candled for cracks prior to setting for incubation?

Were eggs set weekly? Yes

Yes Speed King (Model No. 32) eggcandling lamp, eggs to be incubated were fumigated with formaldehyde

gas. Placed into a NatureForm Incubator (Model No. NMC

- 4000 or NMC-7000)

Eggs should be candled on day 0

When candling was done for fertility? Eggs were candled on Day 11-12
of incubation to determine embryo viability and on Day 21 to determine embryo survival. Speed King (Model No. 32) eggcandling lamp.

Quail: approx. day 11 Ducks: approx. day 14

When the eggs were transferred to the
hatcher? Day 21 Petersime Hatcher (Model No. S6H)

Bobwhite: usually day 21 Mallard: usually day 23

Hatching conditions temperature:

humidity:

photoperiod: 37.2 ± 0.0°C (SD)

– avg. temp. in hatching compartment; 33.3 ± 0.0% (SD)

– avg. wet bulb temp.

77%

Not Reported Eggs were not rotated in the hatcher

Recommended temperature is 39°C (102 °F) Recommended humidity is 70%

Day the hatched eggs were removed and
counted Day 25 or 26 All hatchlings, unhatched eggs, and egg shells were removed from
hatcher.

Eggs for bobwhite should be removed on day 24; for mallard on day 27

Egg shell thickness no. of eggs used:

intervals:

mode of measurement: One egg, when available, from each group

Weekly

Five points around the waist of the shell using a micrometer
calibrated to 0.002 mm. Opened at waist, contents removed, rinsed with water, and air dried for
at least 1 week.

Newly hatched eggs should be collected at least once every two weeks. Thickness of the shell plus membrane
should be measured to the nearest 0.01

mm with 3 - 4 mea surements per shell.

Were egg shells washed and dried for at least 48 hrs. before measuring? Yes, shells were dried for at least
one week

### Data Evaluation Report on the Reproductive Effects of Forchlorfenuron on Avian Species Colinus virginianus PMRA Submission Number {......} EPA MRID Number 49350901



### Page 10 of 15 Parameter Details Remarks

Criteria

Brooding Chambers temperature:

humidity:

photoperiod: 27.3 ± 1.1°C (SD)

– avg. ambient room temp.

21 ± 8% (SD)

– avg. relative humidity

16 hours light per day Thermostats in brooding compartment of each pen were set to maintain a temperature of approx. 38 °C from time of hatching until birds were 14 days old.

Reference chemical, if used N/A; a reference chemical was
not used

2. Observations:

Table 3: Observations.

Parameter Details Remarks

Parameters measured

Parental (mortality, body weight, mean feed consumption)

Egg collection and subsequent development (no. of eggs laid, no. of eggs cracked, shell thickness, no. of eggs set, no. of viable embryos, no. of live 3 week embryos, no. hatched,
no. of 14

- day survivors, average weight of 14 -d old survivors, mortality, gross pathology, others) -Mortality -Signs of toxicity -Body weight
-Feed consumption

- eggs laid - eggs cracked - egg weight - egg shell thickness - eggs set - viable embryos
- live 3 -week embryos - hatchlings - hatchling body weight
- 14 -day-old survivors - 14-day-old survivor body weight Recommended endpoints measured
include:

x Eggs laid/pen

x Eggs cracked/pen

x Eggs set/pen

x Viable embryos/pen

x Live 3-week embryos/pen

x Normal hatchlings/pen

x 14-day-old survivors/pen

x 14-day-old survivors/pen

x Weights of 14-day-old

x survivors (mean per pen)

x Egg shell thickness

x Food consumption (mean per pen)

x Initial and final body weight (mean per pen)

Indicate if the test material was regurgitated None reported

### Data Evaluation Report on the Reproductive Effects of Forchlorfenuron on Avian Species Colinus virginianus PMRA Submission Number {......} EPA MRID Number 49350901



### Page 11 of 15 Parameter Details Remarks

Observation intervals (for various parameters) Birds were observed daily for signs of toxicity and abnormal behavior. Additionally, all offspring were observed daily from hatching until 14 days of age. Adult body weights were measured at test initiation, on weeks 2, 4, 6, 8, and at termination. Feed consumption from each pen was measured weekly throughout
the test. Body weights and food consumption should be measured at least biweekly

Were raw data included? Yes

II. RESULTS AND DISCUSSION:

A. MORTALITY:

No treatment-related mortalities occurre d in the control or treatment groups throughout the test. One incidental death occurred in the highest treatment group. On Day 2 of Week 12 one female was found dead and prior do death was noted to have a head lesion and feather loss. At necropsy the bird was well muscled and had a body weight of 181 grams. Externally, the bird was noted with a laceration at the back of the head and along the dorsal surface of the neck. Internally, a hematoma at the back of the head and neck, and a neck fracture at C1 were noted. A hematoma was also noted on the inner left thigh. The spleen and kidneys were slightly pale, the cecal contents were firm, and the ovary was developing. No other lesions were noted and necropsy of the male pen mate was unremarkable. This single mortality was no t considered to be treatm ent related and the study authors reported a NOAEC of 1000 mg a.i./kg diet

Table 4: Effect of Forchlorfenuron on Mortality of Northern Bobwhite Quail ( Colinus virginianus ).

Mean- Measured and (Nominal)
Concentrations mg a.i./kg diet (ppm a.i.) Observation Period

Week 1 Week 12 Week 20

No. Dead
Male Female No. Dead
Male Female No. Dead
Male Female

Negative Control 0 0 0 0 0 0

170 (160) 0 0 0 0 0 0

435 (400) 0 0 0 0 0 0

1070 (1000) 0 0 0 1 0 1

### Data Evaluation Report on the Reproductive Effects of Forchlorfenuron on Avian Species Colinus virginianus PMRA Submission Number {......} EPA MRID Number 49350901



### Page 12 of 15 B. REPRODUCTIVE AND OTHER ENDPOINTS:

Abnormal Effects/Behavior: No overt signs of toxicity were noted in any treatment group. Incidental clinical observations were made and included those normally asso ciated with injuries and penwear. Such observations included head and foot lesions, ocular injury, an abscess on the head, feather loss, feather picked, nostril picked, and a general pen-worn appearance. Clinical observations, usually associated with the incidental injuries included lameness. Except for these incidental findings, all birds appeared normal throughout the study.

Food Consumption: There were no significant treatment-re lated differences in weekly feed consumption at any treatment level. Overall mean feed consumption ranged from 18 to 19 g/bird/day for all control and treatment groups. The study authors reported a NOAEC of 1000 mg a.i./kg diet.

Body Weights: There were no treatment-related effects on adult body weight for either sex throughout the test. Mean male body weight ranged from 186 to 191 g and from 201 to 207 g at test initiation and termination, respectively. Mean female body weight ranged from 187 to 190 g and from 227 to 234 g at test initiation and termination, respectively. The study authors reported a NOAEC of 1000 mg a.i./kg diet.

The study authors noted significant reductions in hatchling body weight at the 400 and 1000 mg a.i./kg diet treatment groups relative to the control. However, the di fferences were slight (appro ximately 0.5 g; 8%) and were not concentration responsive. Therefore, the study author s did not consider these reductions to be treatmentrelated. The study authors reported a NOAEC of 1000 mg a.i./kg diet.

Reproductive Effects: There were no apparent treatment-r elated effects on any reproductive parameter or eggshell thickness at any treatment level relative to the negative control. The st udy authors reported a NOAEC of 1000 mg a.i./kg diet.

### Data Evaluation Report on the Reproductive Effects of Forchlorfenuron on Avian Species Colinus virginianus PMRA Submission Number {......} EPA MRID Number 49350901



### Page 13 of 15 Table 5: Reproductive and Other Parameters (nominal concentrations; mg/kg-diet)

Parameter Control 100 400 1000 NOAEC/
LOAEC

No. laying pairs 16 16 16 16 N/A

Eggs laid/pen 41 42 40 40 1000/>1000

Eggs laid/hen/day (a) 0.44 0.46 0.43 0.44 1000/>1000

Eggs cracked 6 12 8 15 1000/>1000

Eggs set 552 588 546 514 1000/>1000

Shell thickness (mm ± SD) 0.230±0.012 0.228±0.014 0.221±0.018 0.229±0.013 1000/>1000

Viable embryos 520 509 492 460 1000/>1000

Live 3-week embryos 512 507 485 456 1000/>1000

No. of hatchling/hen 469 478 451 435 1000/>1000

Hatchling weight (g) 5.9 5.8 5.4* (b) 5.5* (b) 1000/>1000

14-day old survivors 437 434 408 380 1000/>1000

14-day old survivors weight (g) 25 24 23 23 1000/>1000

Mean food consumption (g/bird/day) 18 18 19 18 1000/>1000

Weight of females (g; parent)
at test initiation:
at test termination: 190
234 187
230 189
232 189
227 1000/>1000

Weight of males (g; parent)
at test initiation:
at test termination: 191
207 186
201 190
204 190
202 1000/> 1000

Gross pathology None considered treatment-related

(a) Based on 92 days of egg production
(b) Not considered treatment-related * Significantly reduced from control ( p < 0.05).

C. REPORTED STATISTICS:

Adult sex-specific body weight, mean food consumption, and reproductive parameters were analyzed using ANOVA. Dunnett’s multiple comparison procedure was us ed to compare the three treatment means with the control group mean and assess the statistical significance of the observed differences. Sample units were the individual pens within each experimental group, except adult body weights were the sample unit was the individual bird. Percentage data were examined usin g Dunnett’s method following acrsine square root transformations. Percentage data included the following en dpoints: eggs laid of maximum laid, eggs cracked of eggs laid, viable embryos of eggs set, live 3-week embryos of viable embryos, hatchlings of 3-week embryos, 14-d old survivors of hatchings, hatchlings of eggs set, 14-d old survivors of eggs set, hatchlings of maximum set, and 14-d old survivors of maximum set.

### Data Evaluation Report on the Reproductive Effects of Forchlorfenuron on Avian Species Colinus virginianus PMRA Submission Number {......} EPA MRID Number 49350901



### Page 14 of 15 D. VERIFICATION OF STATISTICAL RESULTS:

Statistical Method: The reviewer analyzed eggs laid per, eggs not cracked per egg se t, viable embryos per egg set, live embryos per egg set, hatchlings per egg set, 14-d survivors per egg set, live embryos per viable embryo, hatchlings per live embryo, 14-survivors per normal hatchling, eggshell thickness, and sex-specific adult body weight change. Prior to determining NOAEC values all data were tested for normality using the Shapiro- Wilk’s ( α =0.01) test and for homogeneity of variance using Levene’s test ( α =0.01). Eggs laid per pen, eggshell thickness, and mean food consumption met the assumptions of ANOVA and were analyzed using Dunnett’s test. Male weight gain, hatchling weight, and survivor weight also exhibited a suggestive monotonically decreasing trend and was therefore also analyzed using the Williams (14-d survivor weight, male weight gain) or the Jonckheere-Terpstra (hatchling weight) test. All remainin g endpoints were analyzed using Wilcoxon/Bonferroni Adjustment test. All analyses were conducted using CE TIS statistical software vers ion 1.8.7.12 with database backend settings implemented by EFED on 3/25/2014. All analyses were conducted using the mean-measured concentrations, and the α level was set to 0.05 unless specified otherwise

.
NOAEC: <170 mg a.i./kg diet LOAEC: 170 mg a.i./kg diet Most Sensitive Endpoint(s): 14-d survivor weight

Table 6: Reproductive and Other Parameters (m ean-measured concentrations, mg a.i./kg diet).

Parameter Control 170 435 1070 NOAEC/ LOAEC

Eggs laid/pen 40.6 42.0 39.9 40.2 1070/>1070

Eggs not cracked/eggs laid (%) 99.1 98.1 98.9 97.0 1070/>1070

Viable embryos/eggs set (%) 93.9 87.0 87.2 89.7 1070/>1070

Live embryos/eggs set (%) 92.0 86.2 85.7 89.0 1070/>1070

Hatchlings/eggs set (%) 83.7 82.4 78.6 84.9 1070/>1070

14-d hatchlings/eggs set (%) 76.2 73.2 69.6 75.6 1070/>1070

Live embryos/viable embryos (%) 97.8 99.1 97.3 98.8 1070/>1070

Hatchlings/live embryos (%) 90.8 95.8 90.9 95.7 1070/>1070

14-d hatchlings/normal hatchlings (%) 90.1 88.0 90.0 88.9 1070/>1070

Eggshell thickness (mm) 0.229 0.228 0.221 0.229 1070/>1070

Hatchlings weight (g) 5.75 5.73 5.44 5.47 1070/>1070

14-d survivor weight (g) 25.4 23.6* 23.1* 23.1* <170/170

Mean food consumption (g/bird/day) 17.8 17.7 18.6 17.8 1070/>1070

Male weight gain (g) 16.1 14.9 14.1 11.6 1070/>1070

Female weight gain (g) 44.6 42.4 43.1 38.9 1070/>1070

* Significant reduction relative to the negative control ( William’s test, p < 0.05)

### Data Evaluation Report on the Reproductive Effects of Forchlorfenuron on Avian Species Colinus virginianus PMRA Submission Number {......} EPA MRID Number 49350901



### Page 15 of 15 E. STUDY DEFICIENCIES:

There were no study deficiencies.

F. REVIEWER ’ S COMMENTS:

The reviewer’s results were based on the mean -measured concentrations whereas those of the study authors were based on nominal concentrations. Further, the study authors did not consider the significant reductions in 14-d survivor body weight to be treatment-related. The review er detected significant reductions in 14-d survivor body weight at all treatment levels (7.0-9.1 %) relative to the negative control and these reductions followed a mildlysuggestive dose- dependent pattern. Therefore, the reviewer’s re sults are reported in the Executive Summary and
Conclusions sections of this DER.

Homogeneity of the test substance in the diet was evaluated by collection of six samples from each of the treated diets and one sample from the control diet on Day 0 of Week 1. Samples were collected from the top, middle, and bottom of the left and right sections of the mixing ve ssel. Control and treatment group diet samples were also collected from the feed troughs on Day 7 of Week 1 to assess stability of the test substance under actual test conditions. None of the control samples showed any indic ation of the presence of the test substance or of the presence of co-eluting substances. Means and standard deviations for the three concentrations were 170 ± 5.24, 432 ±5.56, and 1060 ±16.4 mg a.i./kg diet, with coefficien ts of variation of 3.08, 1.29, and 1.55%, respectively.

A sample was also collected from the control and treatment grou p diets during Weeks 2, 3, 4, 8, 12, 16, and 20 of the test to measure/verify test concentrations. Means an d standard deviations were 170 ± 7.90, 435 ± 7.66, and 1070 ± 35.3 mg a.i./kg diet, with coefficients of variation of 4.65, 1.76, and 3.30%, respectively. Mean measured concentrations yielded recoveries of 106 to 109% of nominal.

The in-life portion of the definitive toxicity test was conducted from October 16, 2002 to April 14, 2003.

G. CONCLUSIONS:

This study is scientifically sound but is classified as supplemental because a valid NOAEC was not established. The NOAEC and LOAEC were <170 and 170 mg a.i./kg diet, respectively, based on significant and suggestively dose-dependent inhibitions in 14-d survivor weight at all treatment levels relative to the negative control.

NOAEC: <170 mg a.i./kg diet LOAEC: 170 mg a.i./kg diet Endpoint(s) Affected: 14-d survivor weight

III. REFERENCES:

Merck & Co., Inc. 1991. The Merck Veterinary Manual . Merck & Co. Rahway, NJ. 1832 pp.

Dunnett, C.W. 1955. A Multiple Comparison’s Procedure for Comparing Several Trea tments with a Control. Jour.
Amer. Statis. Assoc . 50: 1096-1121.

Dunnett, C.W. 1964. New Tables for Multiple Comparisons with a Control. Biometrics 20:482-491.