TOXICOLOGY STUDY NO. 87-XE-08JG-10, SEPTEMBER 2008 – APRIL 2010 

PROTOCOL NO. 08JG-80-08-04-02 

TOXICOLOGY PORTFOLIO 

TWO-GENERATION TEST FOR ENDOCRINE DISRUPTION AND REPRODUCTIVE 

TOXICITY IN JAPANESE QUAIL (COTURNIX JAPONICA) USING ENDOSULFAN 

PREPARED BY MICHAEL J. QUINN JR., HEALTH EFFECTS RESEARCH 

PROGRAM 

Approved for public release; distribution unlimited. 

Specialty: 500C, Toxicity Tests 
Use of trademarked name(s) does not imply endorsement by the U.S. Army 

but is intended only to assist in the identification of a specific product. 

ACKNOWLEDGEMENTS 

The authors would like to acknowledge Les Touart for the opportunity to participate in 

this extensive undertaking. We’d also like to thank Patience Browne for her constant 

help and support throughout this endeavor. A warm thank you also goes to Pat Beall, 

Valerie Adams, William Eck, Emily Lent, Larry Williams, Matthew Bazar, Will McCain, 

Lee Crouse, Mark Way, Arthur O’Neill, and Christiana Grim for their assistance during 

the conduct of this study. The authors also appreciate the large amount of endosulfan 

that was used to complete this long term study that was donated by Robert Everich of 

Makhteshim-Agan of North America Inc. This study was done concurrently with two 

other similar ones by U.S. Geological Survey, Patuxent Wildlife Research Center and 

Wildlife International Ltd.; we’ve supported each other throughout each study’s conduct 

in the development and refinement of this test method, and the authors are grateful for 

having had the opportunity to collaborate with such outstanding researchers in this field. 
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 2008-April 2010 

Sponsor

U.S. Environmental Protection Agency, Office of Science Coordination and Policy, 1200 

Pennsylvania Avenue, NW (7203), Washington, DC 20460 

Study Title

Toxicology Study No. 87-XE-08JG-10 

Protocol No. 08JG-80-08-04-02 

Two-Generation Test for Endocrine Disruption and Reproductive Toxicity in Japanese 

Quail (Coturnix japonica) Using Endosulfan 

Authors

Michael J. Quinn, Ph.D. 

Michelle Cook, M.S. 

Terry Hanna 

Alicia Shifflet, A.S. 

Allison Jackovitz, B.S. 

Craig A. McFarland, D.V.M., Ph.D. 

Mark S. Johnson, Ph.D. 

Study Completion Date

April 2010 

Performing Laboratory

U.S. Army Public Health Command 

Portfolio of Toxicology 

Health Effects Research Program (MCHB-IP-THE) 

5158 Blackhawk Road 

Aberdeen Proving Ground, Maryland 21010-5403 

Laboratory Project ID

08JG-80-08-04-02 

 
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 2008-April 2010 

GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT 

The study described in this report was conducted in compliance with Title 40, Code of 

Federal Regulations (CFR), Part 792, Good Laboratory Practice Standards, except for 

the following: 

No deviations from the aforementioned regulation affected the quality or integrity of the 

study or the interpretation of the results. 

____________________________ _________________________ 

Michael J. Quinn Date 

Study Director 

_____________________________ _________________________ 

Mark S. Johnson Date 

Program Manager 

Health Effects Research 
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 2008-April 2010 

Prepared by: 

_____________________________ __________________ 

Michael J. Quinn, Jr. Date 

Biologist, Health Effects Research Program 

Reviewed by: 

____________________________ __________________ 

Craig A. McFarland Date 

Toxicologist, Health Effects Research Program 

Approved by: 

____________________________ __________________ 

Mark S. Johnson Date 

Director, Portfolio of Toxicology
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, 

September 2008-April 2010 

i 

Table of Contents 

Paragraph Page 

1. REFERENCES ................................................................................................ 1 

2. PURPOSE ....................................................................................................... 1 

3. AUTHORITY .................................................................................................... 1 

4. SCOPE ............................................................................................................ 1 

5. BACKGROUND ............................................................................................... 1 

6. MATERIALS AND METHODS ......................................................................... 2 

 6.1. Experimental Design ............................................................................... 2 

 6.2. Data Analysis .......................................................................................... 4 

 6.3. Quality Assurance ................................................................................... 4 

7. RESULTS ........................................................................................................ 5 

 7.1. Eggshell Thickness and Strength ............................................................ 5 

 7.2. Egg Production ........................................................................................ 6 

 7.3. Sperm Measures ..................................................................................... 9 

 7.4. Growth and Body Weight......................................................................... 10 

 

 7.5. Feed Consumption .................................................................................. 16 

 7.6. Cloacal Gland Area ................................................................................. 21 

 

 7.7. Yolk Endosulfan Content ......................................................................... 22 

 7.8. Onset to Puberty ..................................................................................... 23 

 

 7.9. Copulatory Behavior ................................................................................ 24 
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, 

September 2008-April 2010 

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 7.10. Organ Weights ...................................................................................... 29 

 

 7.1. Perivitelline Layer Sperm Penetration ..................................................... 32 

 7.2. Fertility and Hatchability .......................................................................... 33 

8. DISCUSSION .................................................................................................. 34 

9. CONCLUSION ................................................................................................ 36 

10. POINT OF CONTACT .................................................................................... 37 

Appendices 

A - REFERENCES ............................................................................................... A-1 

B - QUALITY ASSURANCE STATEMENT .......................................................... B-1 

C - ARCHIVES AND STUDY PERSONNEL ........................................................ C-1 

D - ANIMAL USE PROTOCOL WITH MODIFICATIONS ..................................... D-1 

E - DEVIATIONS TO THE PROTOCOL..................................................... E-1 

F - DIET PREPARATION REPORT........................................................... F-1 

G - CONTRIBUTING SCIENTIST’S STATISTICAL ANALYSIS REPORT.......... G-1 

H - DRAFT PATHOLOGY REPORT......................................................... H-1 

I - HORMONE ANALYSIS METHODS......................................................... I-1 

J - STATISTICAL ANALYSIS OF HORMONE DATA..................................... J-1 

K - RAW DATA...................................................................................... K-1 

LIST OF FIGURES 

Table 1. List of Critical Events ............................................................................. 4 

Table 2. F0 Eggshell Thickness and Strength ..................................................... 5 

Table 3. F1 Eggshell Thickness and Strength ..................................................... 5 

Table 4. F2 Eggshell Thickness and Strength ..................................................... 6 

Table 5. F0 Egg Production ................................................................................. 7 

Table 6. F1 Egg Production ................................................................................. 7 

Table 7. F2 Egg Production ................................................................................. 8 

Table 8. F0 Sperm Measures .............................................................................. 9 

Table 9. F1 Sperm Measures .............................................................................. 9 

Table 10. F2 Sperm Measures ............................................................................ 10 

Table 11. F0 Adult Body Weight .......................................................................... 11 
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, 

September 2008-April 2010 

iii 

Table 12. F1 Young Growth Measures ................................................................ 12 

Table 13. F1 Adult Body Weight .......................................................................... 13 

Table 14. F2 Young Growth Measures ................................................................ 14 

Table 15. F2 Adult Body Weight .......................................................................... 15 

Table 16. F0 Female Feed Consumption ............................................................ 17 

Table 17. F0 Male Feed Consumption ................................................................ 17 

Table 18. F0 Paired Feed Consumption .............................................................. 18 

Table 19. F1 Female Feed Consumption ............................................................ 18 

Table 20. F1 Male Feed Consumption ................................................................ 19 

Table 21. F1 Paired Feed Consumption .............................................................. 19 

Table 22. F2 Female Feed Consumption ............................................................ 20 

Table 23. F2 Male Feed Consumption ................................................................ 20 

Table 24. F0 Cloacal Gland Area ........................................................................ 21 

Table 25. F1 Cloacal Gland Area ........................................................................ 21 

Table 26. F2 Cloacal Gland Area ........................................................................ 22 

Table 27. F0 Onset to Puberty ............................................................................ 23 

Table 28. F1 Onset to Puberty ............................................................................ 23 

Table 29. F2 Onset to Puberty ............................................................................ 24 

Table 30. F0 Number of Mount Attempts ............................................................ 24 

Table 31. F0 Onset to Mount ............................................................................... 25 

Table 32. F0 Number of Successful Copulations ................................................ 25 

Table 33. F0 Onset to Successful Copulation ..................................................... 25 

Table 34. F1 Number of Mount Attempts ............................................................ 26 

Table 35. F1 Onset to Mount ............................................................................... 26 

Table 36. F1 Number of Successful Copulations ................................................ 27 

Table 37. F1 Onset to Successful Copulation ..................................................... 27 

Table 38. F2 Number of Mount Attempts ............................................................ 28 

Table 39. F2 Onset to Mount ............................................................................... 28 

Table 40. F2 Number of Successful Copulations ................................................ 28 

Table 41. F2 Onset to Successful Copulation ..................................................... 29 

Table 42. F0 Female Organ Weights .................................................................. 29 

Table 43. F0 Male Organ Weights ...................................................................... 30 

Table 44. F1 Female Organ Weights .................................................................. 30 

Table 45. F1 Male Organ Weights ...................................................................... 31 

Table 46. F2 Female Organ Weights .................................................................. 31 

Table 47. F2 Male Organ Weights ...................................................................... 31 

Table 48. Male Gonadal Weights ........................................................................ 32 

Table 49. Ovarian Follicle Counts ....................................................................... 32 

Table 50. Perivitelline Layer Sperm Penetration ................................................. 33 

Table 51. F0 Generation’s Egg Fertility and Hatchability ..................................... 33 

Table 52. F1 Generation’s Egg Fertility and Hatchability ..................................... 34 
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, 

September 2008-April 2010 

iv 

Table 53. F0 Generation Adult Hormone Data .................................................... 34 

Table 54. F1 Generation Adult Hormone Data .................................................... 35 

Table 55. F2 Generation Adult Hormone Data .................................................... 36 

Table 56. Male Japanese Quail Effects Comparison Across Generations .......... 37 

Table 57. Female Japanese Quail Effects Comparison Across Generations ...... 38 

Table 58. Male Copulatory Behavior Comparison Among Generations .............. 38 

LIST OF FIGURES 

Figure 1. Yolk Endosulfan Content ...................................................................... 22 
TOXICOLOGY STUDY NO. 87-XE-08JG-10 

PROTOCOL NO. 08JG-80-08-04-02 

TWO-GENERATION TEST FOR ENDOCRINE DISRUPTION AND REPRODUCTIVE 

TOXICITY IN JAPANESE QUAIL (COTURNIX JAPONICA) USING ENDOSULFAN 

SEPTEMBER 2008 – APRIL 2010 

1 References 

See Appendix A for a listing of references. 

2 Purpose 

The objective of this research is to determine the value added by examining endocrine endpoints in 

a two-generation avian reproductive toxicity test paradigm. 

3 Authority 

This research and assessment addresses part of the environmental safety and occupational health 

(ESOH) objectives outlined in Army Regulation (AR) 200-1, AR 40-5 and AR 70-1, Department of 

Defense Instruction (DODI) 4715.4, and the Army Environmental Research and Technology 

Assessment (AERTA) requirement A (3.3.c). It was conducted as part of the on-going effort, under 

the U.S. Army Environmental Quality Technology, Ordnance Program, Pollution Prevention Pillar, 

under the direction of Research, Development and Engineering Command (RDECOM), 

Environmental Acquisition Logistics & Sustainment Program. 

4 Scope 

This report documents the conduct, findings, and conclusions of using endosulfan to develop and 

assess a two-generation test for endocrine disruption in Japanese Quail. 

5 Background 

The U.S. Environmental Protection Agency (EPA) is implementing an Endocrine Disruptor 

Screening Program comprised of a battery of Tier 1 screening assays and Tier 2 tests. One of the 

Tier 2 tests under development is a two-generation avian assay using Japanese quail (Coturnix 

japonica). This test is designed to characterize dose-response relationships and immediate and 

latent adverse consequences associated with exposure to putative endocrine modulating 

compounds. The avian two-generation assay serves to evaluate potential population effects of 

endocrine modulating compounds on myriad avian species, and represents terrestrial, oviparous 

wildlife species in general. This study will use endosulfan to fully evaluate the proposed test 

method for its ability to detect adverse effects and characterize dose responses in a two-generation 

paradigm, and to develop and optimize a protocol for the avian two-generation assay for 

characterizing endocrine disruptors. This work will help to determine the value added by examining 

endocrine endpoints over two generations in comparison to tests that include only one generation, 

such as the Ecological Effects Guidelines for Avian Reproduction Tests (EPA, 1996).
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 

2008-April 2010 

2 

6 Materials and Methods 

6.1 Experimental Design 

The parental (F0) generation was first exposed to endosulfan (CAS No. 115-29-7), a known 

estrogenic endocrine disrupting chemical, at four weeks of age and throughout the remainder of the 

birds’ lives. Eggs from this generation were incubated and hatched to produce the first generation 

(F1). The F1 generation was exposed to the treatments in ovo (embryonic) through maternal 

deposition and through feed during the entirety of their lives. Eggs from this generation were 

incubated and hatched to produce the second generation (F2). The F2 generation was exposed to 

endosulfan only in ovo through maternal deposition; all feed for the F2 generation was untreated. 

Measures of reproductive behavior, circulating hormone levels, egg quality, reproduction, and 

development were assessed in the P, F1, and F2 generations as described below. Additional 

measures of growth, reproductive maturity, immunocompetence, were assessed in the F1 and F2 

generations as described below. 

Chemical Treatments: 

The sample size for each treatment was 20 males and 20 females to ensure that there were at 

least 12 breeding pairs in the parental generation control group at the end of the treatment period to 

have successfully produced eggs. The lowest reported LC50 from a five-day dietary exposure to 

endosulfan in 14-day old Japanese quail was found to be 1250 ppm (FWS, 1975). The treatment 

levels for this study were 0 (control), 30, 60, 125, and 250 ppm. The highest dose is one-fifth of the 

lowest known LC50 value and assumed not to cause mortality or other severe signs of parental 

toxicity that would have precluded the evaluation of reproductive parameters. The lowest 

concentration should produce no effects on adult health or reproductive measures. Endosulfan 

was supplied by Makhteshim-Agan of North America Inc., Norwalk, CT. The test diets were 

prepared by Wildlife International Ltd., Easton, MD, USA. Homogeneity and concentration 

verifications for endosulfan were made by USACHPPM Directorate of Laboratory Services. Upon 

receipt of the first batch of treated feed, all doses were tested for concentration verification. Once 

all treatment levels were found to be within acceptable ranges, each subsequent batch of treated 

feed required only two randomly selected doses to be tested for concentration verification unless 

the results of these tests were suggestive of further dose levels being tested, according to GLP 

standards. 

Eggs: 

To generate the F0 generation, a minimum of 60 eggs were hatched for each treatment level, to 

provide sufficient numbers of chicks to establish at least 12 breeding pairs (20 pairs was the goal). 

Twenty males and females were used for the study, beginning exposure to the treatments at week 

4. Optimally, a goal of at least 6 eggs were set for hatch for each parental pair of the F0 generation 

to ensure that a minimum of 60 eggs would be hatched for each test level, providing sufficient 

numbers of chicks to establish at least 12 breeding pairs (20 is the goal) per F1 test level. If a 

treatment level produced insufficient eggs to set the minimum, that level would be dropped from 

further analysis in the test. Optimally, a goal of at least 6 eggs was set for each parental pair of the 

F1 generation to ensure that a minimum of 40 eggs would be set and hatched for each treatment 

level. 

During the treatment period, all eggs were collected at least once daily and recorded by hen of 

origin. The egg production rate was determined for each hen. 
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 

2008-April 2010 

3 

After the hatch of each generation, fertility and embryo viability were determined by candling and/or 

dissecting the unhatched eggs. It was decided that candling would not be performed during 

incubation of each generation to prevent delay of embryonic development. Chicks that did not 

hatch within approximately 24 hours of the majority of the other hatching chicks were considered 

unhatched. No assistance was given to chicks during hatching; chicks that remained alive but had 

no clear ability to hatch on their own were humanely removed from the egg and decapitated 

immediately. 

Prior to incubation of subsequent generations, eggs were collected to assess eggshell strength and 

thickness. Eggshell strength was measured by an Egg Force Reader (Orka Technology Ltd.) that 

evaluates the breaking point of an egg’s shell. Eggshell thickness was measured using an Egg 

Shell Thickness Gauge (Orka Technology Ltd.) that measures thickness through ultrasound. 

General Reproductive Fitness Measures and Observations: 

During the treatment period, all birds were observed daily for overt signs of toxicity or other clinical 

signs of illness or trauma; injuries sustained and subsequent treatments were recorded. 

Observations included mortality and clinical signs of toxicity such as lethargy, depression, wing 

droop, ruffled feathers, etc. If an individual or pair was removed during the study due to nontreatment related injury or death, data from all preceding weeks were reevaluated. Clearly 

moribund individuals or those otherwise in severe distress were euthanized by carbon dioxide 

asphyxiation. 

All adult birds underwent necropsy and gross pathology assessment. The wet weight of the brain, 

liver, kidneys, gonads, and thyroid glands were recorded immediately following death/euthanasia. 

Any morphologic abnormalities were recorded in the study’s notebook or necropsy sheets. Adult 

birds that died or were euthanized during the course of the treatment periods were necropsied 

similarly. Histopathology was performed on the brain, right thyroid gland, liver, bursa of Fabricius, 

kidneys, and gonads. Because of the small size of the tissues, the thyroid glands from half of the 

adults were fixed in formalin for histopathologic analyses, while thyroid glands from the remaining 

half were flash frozen and stored for later glandular thyroxine (T4) content. Immediately following 

euthanasia, spermatozoa were released from a weighed section of epididymis to determine percent 

motility and progression and sperm cell concentration per gram epididymis using an IVOS Sperm 

Analyzer, Version 12.1 (Hamilton-Thorne Research, Beverly, MA, USA; methods described by 

McFarland et al., 2008). 

Biochemical/Endocrine Measurements: 

All hormone analyses were conducted at the University of Maryland, Department of Animal and 

Avian Sciences. Glandular T3 and T4 content and serum estradiol were measured by 

radioimmunoassay (RIA) kits (Siemens) and serum testosterone was measured via an Enzyme 

Linked Immuno Sorbent Assay (ELISA) kit (Caymen). Details of their methodologies can be found 

in Appendix I. 

Behavioral measures: 

Initiation of normal male sexual behavior is important for successful reproduction. In reproductively 

mature males of the F1 and F2 generations, trials were preformed when a minimum of 90% of 

control males reached reproductive maturity. The reproductive behaviors measured included: time 

taken for mount attempts to be initiated, number of mount attempts made within the time limit (3 

minutes), time taken to achieve a successful copulation, and number of successful cloacal 
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 

2008-April 2010 

4 

contacts. Males and females of the F1 generation were paired following completion of the 

behavioral measures. Birds from the F2 generation were euthanized upon completion of this 

measure. 

Growth: 

Growth was measured every three days in chicks in the F1 and F2 generations from day of hatch to 

up to at least week 2 of age. Growth measures included body weight, tarsus length, and the length 

from the tip of the beak to the culmen. 

6.2 Data Analysis 

All observational data were recorded in the laboratory notebook and work sheets where applicable. 

Data were analyzed by PHC statistician Shane Hall. Specifics regarding this analysis can be found 

in the Contributing Scientist Report, Appendix G. 

6.3 Quality Assurance 

The U.S. Army Public Health Command (formerly USACHPPM) Good Laboratory Practice Policy—

Policy Memorandum 74 states — All experiments and studies conducted by any element of the 

AIPH Portfolio of Toxicology will be compliant with the applicable Good Laboratory Practice (GLP) 

Standard guideline (USACHPPM 2009). For this study, the test article dictates that the following 

regulation applies: 

Title 40, Code of Federal Regulations (CFR), Part 792, Toxic Substances Control Act (TSCA), 

Good Laboratory Practice Standards: Final Rule 

According to this policy and that these results may be used in regulatory decisions involving the 

EPA, this study was conducted in compliance with TSCA GLP standards and followed the 

appropriate regulatory testing guidelines.. In compliance with the GLP requirements, the AIPH 

Quality Systems Office audited critical phases of this study. A Quality Assurance Statement is 

provided in Appendix B, which provides the dates of these audits along with the audited phases 

and the dates that the results of the audits were reported to Management and the Study Director. 

The additional GLP requirement of identifying the location of the archived information is provided in 

Appendix C as well as the names of personnel contributing to the performance of this study. 

In addition, all Investigators and technicians adhered to The Guide for Care and Use of Laboratory 

Animals (U.S. Department of Health, Education, and Welfare Publication No. NIH 86-23, 1996). 

Table 1. List of Critical Events 

Critical Event Date of Event 

Animal Use Protocol 04/18/2008 

Animals Received (eggs) 11/14/2008 

Study Start 09/03/08 

Experimental Start 12/29/08 

Experimental Completion 04/01/10 
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 

2008-April 2010 

5 

Necropsy Start 07/21/2009 

Necropsy Completion 04/01/2010 

Study Completion 04/2010 

7 Results 

 Eggshell Thickness and Strength. Endosulfan did not cause any differences in eggshell 

thickness for eggs produced by the F0 and F2 generations (Tables 2-4). Eggshell thickness was 

reduced in the two highest dose groups of the F1 generation compared to controls, and eggshells 

from the 250 ppm dose group were also significantly thinner than those from the 30 ppm treatment 

level. Average eggshell strength was significantly higher in the 250 ppm dose group than controls 

for both the F0 and the F1 generations, however endosulfan had no effect on this trait in the F2 

generation. 

Table 2. Eggshell thickness and strength from the F0 generation of Japanese 

quail (Coturnix japonica) exposed to endosulfan. 

F0 Generation ‐ Egg Strength

   Thickness (mm) Strength

   p‐value = 0.788 p‐value = 0.045*

Dose Average Std. Error Average Std. Error

0 0.93 0.07

30 0.890 0.060 1.05 0.06

60 0.981 0.146 1.05 0.05

125 0.846 0.051 1.19 0.05

250 0.869 0.066 1.11 0.06

Table 3. Eggshell thickness and strength from the F1 generation of Japanese 

quail (Coturnix japonica) exposed to endosulfan. 

F1 Generation ‐ Egg Strength

   Thickness (mm) Strength

   p‐value = 0.000* p‐value = 0.016**

Dose Average Std. Error Average Std. Error

0 0.938 0.052 0.96 0.05

30 0.845 0.047 1.02 0.05

60 0.813 0.052 1.04 0.04

125 0.753 0.032 1.09 0.05

250 0.647 0.038 1.19 0.04
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 

2008-April 2010 

6 

Table 4. Eggshell thickness and strength from the F2 generation of Japanese 

quail (Coturnix japonica) exposed to endosulfan. 

F2 Generation ‐ Egg Strength

   Thickness (mm) Strength

   p‐value = 0.924 p‐value = 0.053

Dose Average Std. Error Average Std. Error

0 0.476 0.024 0.89 0.06

30 0.486 0.024 1.28 0.17

60 0.500 0.031 1.03 0.07

125 0.471 0.020 1.00 0.05

250 0.483 0.021 1.14 0.06

 Egg Production. Egg production was not significantly affected in the F0 and F2 generations 

(Tables 5 and 7). In the F1 generation, females from the 250 ppm dose group produced the largest 

number of eggs between the first and second weeks and between the second and third weeks 

(Table 6). 
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 2008-April 2010 

7 

Table 5. Numbers of eggs produced per week by F0 generation Japanese quail (Coturnix japon

endosulfan. Table 6. Numbers of eggs produced per week by F1 generation Japanese quail (Coturnix japonendosulfan. 

Model Term P‐value Dose Week 6 Week 7 Week 8 Week 9 Week 10 Week 11 Week 12 Week 13 Week 14 Week 15 Week 16 Week 

0 0.45 2.00 3.35 4.65 4.50 3.55 3.15 2.90 3.20 2.85 3.00 2.7

30 0.60 2.95 3.80 4.30 4.30 5.20 5.85 6.35 6.65 6.15 6.00 6.260 0.40 2.40 3.80 4.25 4.55 4.95 4.70 5.60 5.10 5.40 5.55 5.6

125 0.05 2.05 4.35 5.45 5.95 5.35 6.20 5.95 5.60 5.75 6.05 6.2250 0.40 2.45 4.15 5.35 5.40 4.75 5.25 5.10 5.75 5.40 4.65 5.2

Overall 0.29 2.35 3.87 4.77 4.90 4.71 4.90 5.00 5.11 4.94 4.90 5.0

Week 0.000

* Statistically signi ficant interaction (p‐value <.05) indicated that egg production change from week to week was signi ficantly di fferent between dose groups.

GEE Estimated Marginal Means

F0Generation Egg Production

GEE Significance Tests

Interaction ‐            

Dose by week

0.000*

Dose 0.051

Model Term P‐value Dose Week 6 Week 7 Week 8 Week 9 Week 10 Week 11 Week 12 Week 13 Week 14 Week 15 Week 16 Week 17 Week 18 Week 19 

0 0.95 2.80 4.45 5.45 5.40 5.70 4.90 5.20 4.50 5.30 5.90 5.90 5.30 5.20 

30 0.10 1.60 3.50 5.50 6.05 6.30 5.55 5.35 5.00 6.05 5.80 5.60 5.75 5.45 60 1.55 3.50 4.95 5.65 5.40 5.70 5.30 3.95 4.50 5.50 5.55 5.40 5.00 4.30 

125 0.25 2.35 4.20 5.60 5.70 6.05 5.55 4.95 4.15 5.00 5.20 5.10 5.95 5.90 250 0.20 3.00 4.95 5.85 5.60 5.45 4.95 5.40 4.35 5.50 5.60 5.25 5.50 5.15 

Overall 0.37 2.56 4.37 5.61 5.62 5.83 5.24 4.94 4.49 5.46 5.60 5.44 5.49 5.17 

Interaction ‐            

Dose by week

0.000*

GEE Significance Tests

* Statistically signi ficant interaction (p‐value <.05) indicated that egg production change from week to week was signi ficantly different between dose groups.

GEE Estimated Marginal Means

F1 Generation Egg Production

Dose 0.539

Week 0.000
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 2008-April 2010 

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Table 7. Numbers of eggs produced per week by F2 generation Japanese quail (Coturnix japon

endosulfan. 

Model Term P‐value Dose Week 6 Week 7 Week 8 Week 9 Week 10 Week 11 Week 12 Week 13 Week 14 Week 15 Week 16 Week 17 We

0 0.35 0.85 3.25 3.95 3.85 4.70 5.95 6.20 5.45 5.70 5.35 5.35 5

30 0.60 1.60 3.30 3.80 4.35 5.15 5.80 6.35 5.30 6.25 5.70 5.80 560 0.45 0.75 3.15 3.85 4.75 5.10 5.65 6.50 5.15 6.05 5.40 5.85 5

125 0.05 0.55 2.95 3.40 4.50 5.40 4.95 6.05 4.90 5.15 4.35 5.00 5250 0.30 1.00 2.55 2.40 3.45 4.00 3.65 4.50 4.25 4.85 4.05 4.30 4

Overall 0.27 0.89 3.03 3.43 4.15 4.84 5.12 5.87 4.99 5.57 4.93 5.22 5

Dose 0.087

Week 0.000

GEE Significance Tests

Interaction ‐            

Dose by week

0.000*

* Statistically signi ficant interaction (p‐value <.05) indicated that egg production change from week to week was significantly different between dose groups.

GEE Estimated Marginal Means

F2 Generation Egg Production
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 

2008-April 2010 

9 

 Sperm Measures. The number of sperm/g epididymis was the only sperm parameter to be 

affected by endosulfan in the F0 generation (Table 8); sperm count in the 125 ppm dose group was 

lower than that of controls. Percent progressive sperm was the only parameter affected in the F1 

generation; sperm from the 250 ppm group tended to move in a progressive direction more often 

than those from the 0, 30, and 60 ppm groups (Table 9). There was also a greater percentage of 

progressively moving sperm from the control and 60 ppm dose groups in the F2 generation (Table 

10). 

Table 8. Percent motility, percent of motile sperm that moved progressively, and 

number of sperm per gram tissue epididymis from the F0 generation of Japanese 

quail (Coturnix japonica) exposed to endosulfan. 

F0 Generation ‐ Sperm

   % Motile % Progressive Sperm/gram

   p‐value = 0.600 p‐value = 0.983 p‐value = 0.033*

Dose Average Std. Error Average Std. Error Average Std. Error

0 93.5 1.2 17.6 1.7 6917.7 385.4

30 84.3 7.9 17.3 2.2 5278.7 786.3

60 89.9 2.1 16.8 1.3 4786.7 709.7

125 86.8 4.0 18.1 2.6 4212.0 610.7

250 88.4 3.3 16.7 1.7 5132.3 443.6

* Tukey Post‐hoc test indicated that dose 125 had a significantly different average sperm per

gram compared to the control group.

Table 9. Percent motility, percent of motile sperm that moved progressively, and 

number of sperm per gram tissue epididymis from the F1 generation of Japanese 

quail (Coturnix japonica) exposed to endosulfan. 

F1 Generation – Sperm

   % Motile % Progressive Sperm/gram

   p‐value = 0.328 p‐value = 0.010* p‐value = 0.338

Dose Average Std. Error Average Std. Error Average Std. Error

0 37.3 8.2 3.8 1.5 443.2 56.6

30 31.6 5.6 3.1 1.0 342.3 55.0

60 45.3 5.9 5.5 1.3 496.2 46.0

125 44.4 5.9 4.5 0.8 427.7 60.4

250 47.8 3.5 9.8 1.9 465.2 53.0

° ID 177 (dose 250) excluded from analysis: recorded 0's for all three measurements

* Tukey post‐hoc test indicated that dose 250 has significantly greater percent progressive

average compared to dose 60, 30, and the control group.
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 

2008-April 2010 

10 

Table 10. Percent motility, percent of motile sperm that moved progressively, and 

number of sperm per gram tissue epididymis from the F2 generation of Japanese 

quail (Coturnix japonica) exposed to endosulfan. 

F2 Generation ‐ Sperm

   % Motile % Progressive Sperm/gram

   p‐value = 0.338* p‐value = 0.014*° p‐value = 0.303*

Dose Average Std. Error Average Std. Error Average Std. Error

0 77.8 4.0 16.0 1.6 5585.6 698.2

30 78.4 3.3 12.3 1.5 5167.8 856.9

60 85.4 1.4 15.6 0.8 6879.8 579.1

125 81.4 1.8 12.2 0.8 5663.6 374.8

250 66.2 6.1 11.6 1.7 5284.2 653.5

* P‐value calculated from Kruskall‐Wallis test ‐ homogeneity of variance assumption violated

for parametric ANOVA test.

     

° Post hoc evaluation indicates that the control group and dose 60 had significantly greater

average percent progressive sperm compared to dose 30, 125, and 250.

 Growth and Body Weight. Chick growth was not assessed in the F0 generation because the 

birds began exposure to endosulfan at 4 weeks of age. Adult female F0 generation body weights 

were not different, however a significant reduction in body weight was observed in males from the 

125 and 250 ppm treatment levels (Table 11). 

 No differences in culmen length were observed for the F1 generation (Table 12), however the 

average tarsus length for week old control chicks was larger than that of the 60 ppm dose group. 

Statistical differences in body weight for the first five weeks of age were observed for the F1 

generation, but these changes were minimal and sporadically distributed among the does groups. 

The dose-by-week interaction was statistically significant for females 6-20 weeks of age in the F1 

generation. All dose-by-week interactions were significant, as there was a great amount of 

variation between doses from week to week. Average body weights were typically highest in 

females in the 250 ppm dose group, with female body weights lowest in the control and 60 ppm 

dose groups. The dose-by-week interaction was not statistically significant for the males, however 

week and dose individually were. The control group’s average body weight was significantly lower 

than the other four dose groups, with the 30 ppm treatment level having a consistently lower 

average body weight than observed at the 60 ppm level. 

 

No significant differences in body weight, tarsus, or culmen measurements were observed in chicks 

from the F2 generation (Table 13). Adult F2 females from the 60 ppm dose group had lower 

average body weights than those from the 30 and 125 ppm dose groups. Control male adults from 

the F2 generation had lower average body weights compared to all other groups. Additionally, 

males at the 125 ppm treatment level had higher average body weights than all other levels. 
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 2008-April 2010 

11 

Table 11. Mean body weight from adult female and male F0 generation Japanese quail (Coturniexposed to endosulfan. 

Factor P‐value Dose Week 6 Week 7 Week 8 Week 9 Week 10 Week 11 Week 12 Week 13 Week 14 Week 15 Week 

0 105.1 113.9 124.7 130.0 129.8 132.1 131.5 136.3 137.4 138.2 134.

30 108.9 120.5 125.5 125.7 129.7 133.3 136.4 138.2 138.2 139.5 139.60 103.7 115.2 125.3 127.6 130.5 133.7 131.5 134.6 136.9 137.0 138.

125 104.3 116.6 126.7 129.5 131.9 131.5 135.9 135.9 134.0 137.7 140.250 105.1 117.6 125.4 130.3 133.7 132.3 134.3 135.0 141.9 139.4 133.

Overall 105.4 116.7 125.5 128.6 131.1 132.5 133.9 136.0 137.7 138.3 137.

Factor P‐value Dose Week 6 Week 7 Week 8 Week 9 Week 10 Week 11 Week 12 Week 13 Week 14 Week 15 Week 

0 98.2 103.6 105.2 107.6 107.8 109.5 108.0 109.2 110.3 111.5 111.

30 99.5 104.8 106.4 106.7 107.8 110.2 107.9 111.3 112.7 112.0 111.60 99.2 103.9 105.6 107.3 109.2 109.9 107.9 108.6 112.0 111.1 113.

125 100.0 103.1 103.2 104.0 105.0 106.6 104.4 105.8 102.3 109.5 111.250 98.0 1010.9 103.5 105.4 106.8 108.0 106.6 108.3 108.6 108.8 110.

Overall 99.0 103.4 104.8 106.2 107.3 108.8 106.9 108.6 109.2 110.6 111.

Week 0.000*

Interaction ‐ 

dose by week

0.999

Dose 0.000*

Week 0.000**

F0Generation Female Body Weights

F0Generation Male Body Weights

Significance Tests Estimated Marginal Means

Significance Tests Estimated Marginal Means

Interaction ‐ 

dose by week

0.917

Dose 0.161

* Tukey post‐hoc tests indicate that weekly averages are significantly di fferent from each other.

* Tukey post‐hoc tests indicate that dose 125 had significant lower body weight compared to the control group, dose 30, and dose 60. Dose 250 had a significantly low

60.** Tukey post‐hoc tests indicate that weekly averages are significantly di fferent from each other.
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 

2008-April 2010 

12 

Table 12. Mean body weight from young female and male F1 generation 

Japanese quail (Coturnix japonica) exposed to endosulfan. 

Week 1 Week 2 Week 3

Female Male Female Male Female Male

p‐value = .13 p‐value = .00 N/A* p‐value = .00 p‐value = .00 p‐value = .30 p‐value = .58 p‐value = .35 p‐value = .01

0 13.5 35.0 54.3 78.2 75.5 97.3 91.1 113.9 94.2

30 14.2 36.9 80.0 76.4 75.0 93.1 95.6 110.3 102.9

60 15.8 26.7 51.3 66.9 80.3 100.1 93.1 118.5 98.9

125 18.4 34.8 58.9 79.2 67.9 99.4 95.3 115.5 101.5

250 18.3 35.2 58.0 80.7 76.4 102.1 93.2 113.4 103.6

p‐value = .33 p‐value = .00 p‐value = .63 p‐value = .61 p‐value = .73 p‐value = .17 p‐value = .60 p‐value = .22 p‐value = .65

0 18.1 22.6 20.4 32.7 32.1 33.3 33.1 33.6 33.1

30 19.0 21.3 21.7 32.8 32.1 33.3 32.7 33.4 32.7

60 19.3 19.0 21.4 32.4 32.5 33.3 33.0 33.4 32.8

125 19.9 20.1 21.3 33.3 32.0 33.3 33.0 33.2 33.0

250 19.3 20.5 20.6 32.5 32.1 34.3 33.3 34.1 33.2

p‐value = .14 p‐value = .09 p‐value = .53 p‐value = .96 p‐value = .52 p‐value = .05 p‐value = .41 p‐value = .48 p‐value = .11

0 3.7 6.0 7.4 6.8 6.8 7.6 7.2 7.4 6.9

30 3.9 6.5 7.1 6.9 6.7 7.7 7.1 7.2 7.2

60 3.8 6.3 7.5 6.9 7.0 7.4 7.0 7.4 6.9

125 4.1 6.3 7.4 6.9 6.8 7.2 7.3 7.4 7.4

250 4.1 5.8 7.0 6.8 6.9 7.5 7.2 7.1 7.4

* Dose 30 only had 1 observa tion.  Dose 30 va riance was coul d not be calcula ted s o the ANOVA tes t was unable to be executed.

Dose

F1 Generation Tarsus, Culmen, and Body Weight Averages

Tarsus

Culmen

Week 4 Week 5 Week 6

Body Weight
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 2008-April 2010 

13 

Table 13. Mean body weight from adult female and male F1 generation Japanese quail (Coturniexposed to endosulfan. 

Factor P‐value Dose Week 7 Week 8 Week 9 Week 10 Week 11 Week 12 Week 13 Week 14 Week 15 Week 16 Week 17 Week 18 We

0 118.4 126.5 128.5 127.5 130.0 131.3 130.1 132.1 135.9 134.7 134.3 134.1 1

30 124.2 129.0 136.3 134.9 135.1 136.2 136.6 138.1 136.3 140.9 138.3 140.1 160 138.2 126.8 131.9 130.7 131.2 131.9 126.8 131.0 131.0 139.6 133.3 125.0 1

125 126.5 132.5 133.7 135.1 134.8 136.0 136.8 131.8 137.0 139.3 138.7 142.0 1250 130.6 132.6 136.1 137.1 133.6 138.5 139.1 139.3 142.4 142.4 139.3 141.5 1

Overall 127.5 129.4 133.3 133.0 132.9 134.8 133.9 134.5 136.5 139.4 136.8 136.5 1

Factor P‐value Dose Week 7 Week 8 Week 9 Week 10 Week 11 Week 12 Week 13 Week 14 Week 15 Week 16 Week 17 Week 18 We

0 101.9 103.7 105.1 105.1 106.5 105.9 104.6 106.8 111.7 108.3 109.0 112.2 1

30 103.8 107.4 108.7 107.9 109.2 107.0 107.0 111.8 111.3 111.1 112.0 112.2 160 106.2 108.8 110.8 110.4 111.5 111.3 110.0 111.6 113.9 114.5 115.5 109.9 1

125 107.4 109.1 109.9 109.3 110.7 109.7 108.9 114.4 111.2 111.8 112.5 112.2 1250 104.8 107.7 108.1 107.6 108.4 108.3 110.2 109.5 111.2 111.7 112.1 112.5 1

Overall 104.8 107.3 108.5 108.0 109.2 108.4 108.1 110.6 111.8 111.5 112.2 111.8 1

Week 0.000**

F1Generation Male Body Weights

F1Generation Female Body Weights

Week 0.000

Significance Tests

Interaction ‐ 

dose by week

0.996

Significance Tests

Interaction ‐ 

dose by week

0.005*

Dose 0.000

Estimated Marginal Means

Estimated Marginal Means

Dose 0.000*

* Statistically significant interaction indicated that body weight change from week to week was signi ficantly different between the five dose groups.* Tukey post‐hoc tests indicated that the control group's body weight was significantly lower than the other four dose groups. Dose 30 also ahd a significantly lower b** Tukey post‐hoc tests indicate that weekly averages are significantly di fferent from each other.
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 

2008-April 2010 

14 

Table 14. Mean body weight from young female and male F2 generation 

Japanese quail (Coturnix japonica) exposed to endosulfan. 

Dose Week 1 Week 2 ‐ A Week 2 ‐ B Week 3 ‐ A Week 3 ‐ B Week 4 ‐ A Week 4 ‐ B Week 5 ‐ A Week 5 ‐ B

p‐value = .91 p‐value = .94 p‐value = .47 p‐value = .57 p‐value = .95 p‐value = .77 p‐value = .83 p‐value = .65 p‐value = .78

0 7.6 16.2 24.8 34.6 46.9 57.0 70.3 78.7 81.8

30 7.2 16.9 25.7 36.2 47.7 60.3 72.3 80.6 84.0

60 7.7 17.2 26.6 37.4 48.2 59.6 72.0 79.2 83.2

125 7.7 17.1 24.6 36.8 47.7 60.3 73.1 82.5 84.6

250 7.6 16.3 23.9 36.0 46.6 58.4 74.0 82.8 86.0

p‐value = .80 p‐value = .26 p‐value = .28 p‐value = .31 p‐value = .14 p‐value = .39 p‐value = .16 p‐value = .26 p‐value = .74

0 15.6 17.8 20.9 23.0 27.2 28.0 30.3 31.0 32.7

30 15.8 19.4 21.4 23.4 26.9 29.7 31.5 31.8 32.8

60 16.1 19.1 21.3 24.1 28.3 29.7 31.7 31.9 32.3

125 15.8 19.8 19.0 23.1 27.1 28.6 31.8 32.8 32.3

250 16.0 18.9 20.1 22.4 26.0 28.5 31.5 32.4 33.1

p‐value = .37 p‐value = .70 p‐value = .43 p‐value = .76 p‐value = .57 p‐value = .56 p‐value = .26 p‐value = .79 p‐value = .82

0 3.8 4.7 5.6 6.3 6.7 7.3 7.4 7.9 7.8

30 3.7 4.7 5.6 6.5 6.6 7.4 7.8 8.0 7.7

60 3.9 4.9 5.7 6.6 5.8 7.7 7.5 8.1 7.9

125 3.7 4.9 5.9 6.3 5.9 7.3 7.4 7.9 7.8

250 4.0 4.8 5.5 6.6 6.6 7.4 7.2 7.9 7.7

Body Weight

Tarsus

Culmen

F2 Generation Tarsus, Culmen, and Body Weight Averages
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 2008-April 2010 

15 

Table 15. Mean body weight from adult female and male F1 generation Japanese quail (Coturniexposed to endosulfan. 

Factor P‐value Dose Week 6 Week 7 Week 8 Week 9 Week 10 Week 11 Week 12 Week 13 Week 14 Week 15 Week 16 We

0 100.9 108.8 124.3 134.5 133.5 132.3 137.8 137.9 140.3 141.6 138.3 1

30 106.7 116.0 124.2 130.5 133.9 135.6 137.6 138.5 139.7 143.2 139.8 160 102.1 108.2 124.5 131.5 131.6 132.5 135.9 138.3 138.5 140.0 136.5 1

125 103.5 112.6 125.2 129.6 136.1 139.3 139.7 140.1 140.4 142.4 139.6 1250 108.1 115.7 125.8 128.1 131.6 133.4 135.9 137.7 139.3 142.0 140.0 1

Overall 104.2 112.2 124.8 130.8 133.3 134.6 137.5 138.5 139.6 141.8 138.8 1

Factor P‐value Dose Week 6 Week 7 Week 8 Week 9 Week 10 Week 11 Week 12 Week 13 Week 14 Week 15 Week 16 We

0 94.3 98.1 103.4 104.4 104.7 106.1 108.5 108.5 108.7 109.4 109.8 1

30 100.4 104.2 107.3 109.4 109.5 110.7 112.6 112.7 113.4 114.1 114.1 1

60 97.6 101.9 106.0 108.0 108.7 110.0 110.8 112.0 111.4 112.9 112.9 1

125 102.8 106.9 109.7 111.7 113.4 113.6 115.1 115.6 114.3 115.1 114.3 1250 101.0 103.6 104.1 107.2 108.6 110.2 111.9 113.0 111.2 112.8 113.3 1

Overall 99.2 102.9 106.1 108.1 109.0 110.1 111.8 112.3 111.8 112.8 112.9 1

Significance Tests

Interaction ‐ 

dose by week 0.948

Estimated Marginal Means

Week 0.000**

F2Generation Male Body Weights

Significance Tests Estimated Marginal Means

* Tukey post‐hoc tests indicated that dose 60 had a signi ficantly lower body weight compared to dose 30 and dose 125.

** Tukey post‐hoc tests indicate that weekly averages are significantly di fferent from each other.

** Tukey post‐hoc tests indicate that weekly averages are significantly di fferent from each other.

* Tukey post‐hoc tests indicate that the control group had a significantly lower body weight compared to the other four dose groups. Dose 125 had a signi ficantly higher body 

groups.

F2 Generation Female Body Weights

Interaction ‐ 

dose by week

1.000

Dose 0.000*

Week 0.000**

Dose 0.003*
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 

2008-April 2010 

16 

 Feed Consumption. While same-sex paired, the juvenile control males and females from the F0 

generation had greater increases in food consumption than all other dose groups (Tables 16 and 

17). When males and females were paired as adults, the control, 60, and 250 ppm dose groups 

consumed less food than pairs from the 30 and 125 ppm dose groups (Table 18). For the F1 

generation, juvenile females from the 250 ppm dose group consumed more food than the other 

doses (Table 19); no differences were found with juvenile male feed consumption (Table 20). F1 

generation adult male/female pairs from the 125 ppm treatment level had a lower average feed 

consumption than those from the control and 250 ppm levels only during the first week of being 

paired (Table 21). F2 females from the control dose group had the highest average for feed 

consumption at ten weeks of age and greater, and no significant differences were observed in 

males (Tables 22 and 23). 
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 2008-April 2010 

17 

Table 16. Mean weekly (from week 5 through 10) feed consumption for F0 generation female Ja(Coturnix japonica) exposed to endosulfan. 

F0 Generation Female Food Consumption (Weeks 5‐10)

Significance Tests Estimated Marginal Means

Factor P‐value Dose Week 5 Week 6 Week 7 Week 8 Week 9 Week 10 

Interaction ‐ dose

by week 0.003 0 113.1 114.9 138.7 154.6 166.7 199.8 

30 132.0 134.8 146.1 146.0 156.8 182.5 

Dose 0.000 60 117.6 121.9 154.3 154.0 154.3 196.5 

125 123.1 141.1 146.7 187.0 191.2 187.2 

Week 0.000

250 125.4 145.6 147.8 175.5 175.4 186.7 

Overall 122.2 131.6 146.7 163.6 168.9 190.5

Table 17. Mean weekly (from week 5 through 10) feed consumption for F0 generation male Jap(Coturnix japonica) exposed to endosulfan. 

F0 Generation Male Food Consumption (Weeks 5‐10)

Significance Tests Estimated Marginal Means

Factor P‐value Dose Week 5 Week 6 Week 7 Week 8 Week 9 Week 10 

Interaction ‐ dose

by week 0.111 0 107.4 121.2 131.7 123.1 121.4 136.9 

30 109.5 117.3 122.0 107.4 106.8 118.3 

Dose 0.000 60 113.5 113.2 121.3 115.9 116.1 118.8 

125 112.9 111.0 112.1 118.6 109.7 116.4 

Week 0.000 250 106.6 111.4 114.9 117.9 114.5 121.1 

Overall 110.0 114.8 120.4 116.6 113.7 122.2

* ID 69 ‐ dose 30 data points removed for week 9 and week 10 due to low food consumption results.
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 2008-April 2010 

18 

Table 18. Mean weekly (from week 11 through 18) feed consumption for F0 generation female aquail (Coturnix japonica) pairs exposed to endosulfan. 

Factor P‐value Dose Week 11 Week 12 Week 13 Week 14 Week 15 Week 16 Week 17 We

0 154.0 290.9 305.4 306.1 282.1 293.6 289.1 11

30 160.4 325.7 324.9 326.5 323.6 302.4 316.2 1260 159.0 300.9 279.2 330.5 292.1 289.9 295.9 12

125 163.4 320.2 320.1 323.6 317.9 313.6 315.7 12250 161.8 299.5 291.0 305.5 292.0 292.4 318.6 12

Overall 159.7 307.5 304.1 318.4 301.5 298.4 307.1 12

F0 Generation Food Consumption (Week 11‐18)

Significance Tests Estimated Marginal Means

Interaction ‐ 

dose by week

0.6400.000

Week 0.000

Dose

Table 19. Mean weekly (from week 7 through 12) feed consumption for F1 generation female Ja(Coturnix japonica) exposed to endosulfan. 

Factor P‐value Dose Week 7 Week 8 Week 11 Week 12 Overall

0 74.6 126.1 150.1 196.8 131.3

30 78.4 123.4 151.5 190.5 129.960 102.9 131.6 147.0 199.8 137.4

125 89.5 124.7 143.4 202.1 133.8250 103.3 145.5 163.1 215.1 149.5

Overall 89.7 130.3 151.0 200.8

Interaction ‐ 

dose by week 0.069

Dose 0.000

Week

* Id 119, 12, 299, 163, and 163 datapoints removed for week 7 due to low food consumption results.

0.000

F1Generation Female Food Consumption (Weeks 7‐12)

Significance Tests Estimated Marginal Means
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 2008-April 2010 

19 

Table 20. Mean weekly (from week 7 through 12) feed consumption for F1 generation male Jap(Coturnix japonica) exposed to endosulfan. 

Factor P‐value Dose Week 7 Week 8 Week 11 Week 12 Overall

0 79.4 90.1 95.6 124.1 97.6

30 71.9 91.6 93.3 117.9 94.060 68.5 97.5 102.7 124.1 99.1

125 70.5 95.5 102.4 126.1 97.1250 74.7 95.8 100.5 120.3 97.7

Overall 73.0 94.5 99.1 122.5

Week 0.000

Dose 0.188

F1Generation Male Food Consumption (Weeks 7‐12)

Significance Tests Estimated Marginal Means

Interaction ‐ 

dose by week

0.157

Table 21. Mean weekly (from week 13 through 21) feed consumption for F1 generation female aquail (Coturnix japonica) pairs exposed to endosulfan. 

Factor P‐value Dose Week 13 Week 14 Week 15 Week 16 Week 17 Week 18 Week 19 Week 2

0 179.3 130.9 346.4 293.4 315.3 309.1 286.7 327.8 

30 161.7 142.6 338.3 263.2 291.0 298.4 278.4 270.8 60 173.4 138.4 329.1 266.5 290.9 286.1 268.1 331.4 

125 161.0 130.9 323.3 246.9 297.9 294.5 294.3 279.7 250 172.8 131.8 336.8 281.8 301.5 308.1 304.2 316.7 

Overall 169.6 134.9 334.7 270.4 299.3 299.2 286.3 305.3 

0.001

Estimated Marginal Means

F1 Generation Food Consumption (Week 13‐21)

Significance Tests

Interactio

n ‐ dose

0.559

DoseWeek 0.000
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 2008-April 2010 

20 

Table 22. Mean weekly (from week 6 through 18) feed consumption for F2 generation female Ja(Coturnix japonica) exposed to endosulfan. 

Factor P‐value Dose Week 6 Week 7 Week 8 Week 9 Week 10 Week 11 Week 12 Week 13 Week 14 Week 15 Wee

0 91.1 114.7 138.7 98.8 130.7 195.8 219.1 243.9 193.7 231.8 214

30 103.4 120.9 125.0 112.4 134.6 183.6 197.7 216.3 202.2 212.5 11160 73.5 108.8 128.3 91.1 136.4 193.9 207.0 236.3 200.7 213.2 107

125 94.5 116.0 125.3 91.7 139.3 169.6 178.0 222.6 183.3 198.7 195250 84.5 106.8 123.9 100.8 127.4 187.3 202.5 249.1 193.9 202.0 179

Overall 90.0 113.4 128.2 98.9 133.7 186.2 200.8 233.6 194.7 211.6 161

F2Generation Female Food Consumption (Week 6‐18)

Estimated Marginal Means

Week 0.000

Significance Tests

Interaction ‐ 

dose by week

0.000

Dose 0.000

* Id 305 (week 9), 215 (week 9), 86 (week 16), 73 (week 16), 84 (week 16) datapoints removed due to inaccurate food consumption results

Table 23. Mean weekly (from week 6 through 18) feed consumption for F2 generation male Jap(Coturnix japonica) exposed to endosulfan. 

Factor P‐value Dose Week 6 Week 7 Week 8 Week 9 Week 10 Week 11 Week 12 Week 13 Week 14 Week 15 We

0 97.8 120.8 122.0 114.6 104.6 121.3 120.9 196.0 119.1 133.7 1

30 98.9 115.4 119.6 122.4 107.7 113.6 110.4 200.1 122.8 129.9 160 89.2 121.7 127.2 123.9 115.2 122.1 130.0 204.1 123.9 127.6 1

125 94.2 125.0 129.3 122.0 122.5 120.1 116.1 193.8 119.1 126.0 1250 96.6 116.8 117.3 103.9 132.5 122.1 124.5 198.1 122.3 124.0 1

Overall 95.3 119.9 123.1 117.4 116.5 119.8 120.3 198.4 121.4 128.2 1

F2 Generation Male Food Consumption (Week 6‐18)

Estimated Marginal Means

Dose 0.077

Week 0.000

Significance Tests

Interaction ‐ 

dose by week

0.003

* ID 193 datapoint removed from week 7 due to inaccurate food consumption result.
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 

2008-March 2010 

21 

 Cloacal Gland Area. Control males from the F0 and F1 generations had larger cloacal glands 

during development, however they were only statistically larger than those from the 60 ppm dose 

group in the F0 generation and the 30 ppm dose group in the F1 generation (Tables 23 and 24). 

Endosulfan had no effect on cloacal gland size in the F2 generation (Table 25). 

Table 24. Mean cloacal gland area (mm) from F0 generation male Japanese quail 

(Coturnix japonica) exposed to endosulfan. 

Factor P‐value Dose Week 6 Week 7 Overall

0 113.4 164.4 138.9

30 75.4 118.6 97.0

60 70.0 114.2 92.1

125 97.7 165.0 131.4

250 84.1 136.7 110.4

Overall 88.1 169.8

F0 Generation Cloaca

Interaction ‐ 

dose by week

Dose

Week

0.230

0.013*

0.000**

Significance Tests

* Tukey post‐hoc tests indicate that the control group had a signi ficantly greater CG

area compared to dose 60.

Estimated Marginal Means

** Week 7 had significantly greater CG area compared to week 6.

Table 25. Mean cloacal gland area (mm) from F1 generation male Japanese quail 

(Coturnix japonica) exposed to endosulfan. 

Factor P‐value Dose Week 6 Week 7 Overall

0 139.7 189.3 164.5

30 89.2 157.0 123.1

60 150.7 177.2 163.9

125 109.5 173.5 141.5

250 119.6 160.8 140.2

Overall 121.7 171.5

F1 Generation Cloaca

Significance Tests Estimated Marginal Means

Interaction ‐ 

dose by week

0.588

Dose 0.017*

Week 0.000**

** Week 7 had a significantly greater CG area average compared to week 6.

* Tukey post‐hoc tests indicated that the control group had a significantly greater CG

area compared to dose 30.
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 

2008-March 2010 

22 

Table 26. Mean cloacal gland area (mm) from F2 generation male Japanese quail 

(Coturnix japonica) exposed to endosulfan. 

Factor P‐value Dose Week 6 Week 7 Week 8 Overall

0 104.0 151.5 205.3 153.6

30 100.4 160.8 218.3 159.8

60 76.8 152.8 210.0 146.5

125 97.4 148.6 225.8 157.3

250 74.7 148.9 201.7 141.8

Overall 90.6 152.5 212.2

Dose 0.905

Interaction ‐ 

dose by week

* All weeks were signi ficantly di fferent from each other.

Significance Tests Estimated Marginal Means

F2 Generation Cloaca

Week 0.000*

0.785

 Yolk Content. Figure 1 shows endosulfan concentrations in yolk samples collected from F0 

generation eggs. A dose-dependent increase is apparent for both the α- (endosulfan I) and βisomers (endosulfan II). 

Figure 1. Yolk concentrations of endosulfan I and II from eggs collected from the 

F0 generation of Japanese quail (Coturnix japonica) that were exposed to 

endosulfan via feed. 
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 

2008-March 2010 

23 

 Onset to Puberty. Onset to puberty was unaltered by endosulfan in the F0 generation (Table 

27). Although the p-value for male onset was slightly less than 0.05, there were no significant 

differences between treatment levels after accounting for multiple testing. Females in the F1 

generation from the 30 ppm dose group had significantly longer times to onset of reproductive 

maturity than females from the control and 60 ppm dose groups. Males from all dose groups 

exhibited a similar pattern of onset to puberty (Table 28). No differences in onset to puberty were 

observed for either gender in the F2 generation (Table 29). 

Table 27. Mean onset to reproductive maturation in male and females Japanese 

quail (Coturnix japonica) of the F0 generation that were exposed to endosulfan. 

Table 28. Mean onset to reproductive maturation in male and females Japanese 

quail (Coturnix japonica) of the F1 generation that were exposed to endosulfan.

Dose Average Std. Error Average Std. Error

0 57.2 1.68 44.7 0.47

30 56.1 1.81 46.8 0.67

60 56.6 1.70 45.9 0.64

125 56.3 1.05 44.7 0.51

250 56.3 1.44 45.8 0.59

P‐value = .991 P‐value = .044

Females Males

F0 Generation Onset to Puberty (days)

Dose Average Std. Error Average Std. Error

0 49.6 1.44 43.2 2.47

30 55.4 1.31 46.0 1.86

60 49.9 1.59 41.6 1.92

125 54.3 1.27 44.3 1.84

250 53.7 1.17 46.5 1.92

Females Males

P‐value = .007* P‐value = .405

* Tukey post‐hoc tests indicate that dose 30 has

signi ficantly longer onset to puberty compared to the

control group and dose 60.

F1Generation Onset to Puberty (days)
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 

2008-March 2010 

24 

Table 29. Mean onset to reproductive maturation in male and females Japanese 

quail (Coturnix japonica) of the F0 generation that were exposed to endosulfan. 

 Copulatory Behavior. For the F0 generation, males from the control and 125 ppm groups had 

significantly greater numbers of mount attempts compared to males from the 30, 60, and 250 ppm 

groups in the first of three trials (Table 30). In the second trial, males from the 125 ppm dose group 

had a significantly greater number of mount attempts compared to the control, 30, and 60 ppm 

males. No other differences in copulatory behavior were observed for F0 generation males. Time 

to initiate mount (Table 31), number of copulations per trial (Table 32), and time until first copulation 

(Table 33) were unaffected by endosulfan in the F0 generation. 

Table 30. Mean number of mount attempts initiated within a 3 minute time period 

per trial by male Japanese quail (Coturnix japonica) from the F0 generation that 

were exposed to endosulfan. 

Dose Average Std. Error Average Std. Error

0 53.2 1.94 48.1 1.32

30 54.4 2.18 48.7 1.46

60 55.7 1.72 48.1 1.26

125 57.3 1.81 47.1 1.23

250 55.3 2.66 48.1 1.24

F2 Generation Onset to Puberty (days)

Females Males

P‐value = .680 P‐value = .933

Dose

Average number

of Mounts

95% β Confidence

Interval* P‐value Average number

of Mounts

95% β Confidence

Interval* P‐value Average number

of Mounts

95% β Confidence

Interval* P‐value

0 6.60 4.95 4.40

30 4.60 .53 ‐ .91 0.01 4.35 .66 ‐ 1.18 0.39 5.00 .81 ‐ 1.44 0.38

60 4.20 .48 ‐ .84 0.00 4.75 .72 ‐ 1.28 0.78 5.35 .89 ‐ 1.58 0.17

125 6.55 .78 ‐ 1.26 0.95 6.90 1.07 ‐ 1.81 0.01 5.21 .92 ‐ 1.61 0.25

250 4.40 .51 ‐ .87 0.00 5.55 .85 ‐ 1.48 0.41 4.75 .81 ‐ 1.44 0.61

* Confidence Intervals reflect comparison to control group (ppm = 0).  Ppm levels with a confidence interval not including 1.00 is considered to be significantly different

F0Generation ‐ Number of Mounts

Trial 1

Omnibus Likelihood ratio p‐value = .000

Trial 2

Omnibus Likelihood ratio p‐value = .006

Trial 3

Omnibus Likelihood ratio p‐value = .676
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 

2008-March 2010 

25 

Table 31. Mean amount of time (s) from start of test until first mount attempt per 

trial by male Japanese quail (Coturnix japonica) from the F0 generation exposed 

to endosulfan. 

Table 32. Mean number of successful copulations within a 3 minute period per 

trial completed by Japanese quail (Coturnix japonica) from the F0 generation that 

were exposed to endosulfan. 

Table 33. Mean amount of time (s) from start of test until first successful 

copulation per trial by male Japanese quail (Coturnix japonica) from the F0 

generation exposed to endosulfan. 

Dose

0 38.5 12.7 90% 21.9 11.8 90% 21.5 11.8 90%

30 58.1 14.6 80% 27.8 11.9 95% 14.7 8.6 95%

60 77.6 14.8 80% 28.6 10.0 95% 22.5 9.2 95%

125 26.5 9.3 95% 18.7 9.2 95% 15.7 9.0 95%

250 47.9 13.7 85% 16.5 6.3 100% 10.8 4.1 100%

F0 Generation ‐ Time to first Mount

Trial 1 Trial 2 Trial 3

Log rank p‐value = .069 Log rank p‐value = .830 Log rank p‐value = .688

Average time to

first mount

Standard

Error

Mounted

Pairs

Average time to

first mount

Standard

Error

Mounted

Pairs

Average time to

first mount

Standard

Error

Mounted

Pairs

Trial 2

Dose

Average number

of Copulations

95% β Confidence

Interval* P‐value Average number

of Copulations

95% β Confidence

Interval* P‐value Average number

of Copulations

95% β Confidence

Interval* P‐value

0 0.30 0.42 0.50

30 0.35 .39 ‐ 3.47 0.78 0.50 .49 ‐ 3.01 0.72 0.65 .57 ‐ 2.97 0.53

60 0.10 .07 ‐ 1.65 0.18 0.15 .09 ‐ 1.34 0.13 0.20 .13 ‐ 1.28 0.12

125 0.20 .19 ‐ 2.36 0.53 0.58 .55 ‐ 3.42 0.49 0.55 .47 ‐ 2.59 0.83

250 0.30 .32 ‐ 3.10 1.00 0.40 .36 ‐ 2.53 0.92 0.55 .47 ‐ 2.59 0.83

* Confidence Intervals reflect comparison to control group (ppm = 0).  Ppm levels with a confidence interval not including 1.00 is considered to be significantly different

Omnibus Likelihood ratio p‐value = .205 Omnibus Likelihood ratio p‐value = .226

F0Generation ‐ Number of Copulations

Trial 1

Omnibus Likelihood ratio p‐value = .458

Trial 3

Dose

0 154.7 12.7 20% 121.6 16.5 42% 115.8 16.6 45%

30 144.0 12.5 30% 107.9 16.4 50% 110.5 17.6 45%

60 170.6 7.3 10% 159.8 11.2 15% 151.8 13.0 20%

125 150.3 13.4 20% 120.3 17.4 47% 122.1 17.3 40%

250 139.9 15.7 26% 124.3 17.1 35% 92.9 17.7 55%

Copulated

pairs

F0Generation ‐ Time to first Copulation

Log rank p‐value = .624 Log rank p‐value = .220 Log rank p‐value = .248

Trial 1 Trial 2 Trial 3

Average time

to first

copulation

Standard

Error

Average time

to first

copulation

Standard

Error Copulated

pairs

Average time

to first

copulation

Standard

Error Copulated

pairs
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 

2008-March 2010 

26 

No significant differences were observed in either of three trials for number of mount attempts, elapsed 

time until first copulatory attempt, and number of successful copulations in male Japanese quail from the 

F1 generation (Tables, 34-36). In the last of three trials, males from the 125 ppm dose group took 

significantly longer to achieve a successful copulation than males from the 30 ppm dose group (Table 

37). 

Table 34. Mean number of mount attempts initiated within a 3 minute time period 

per trial by male Japanese quail (Coturnix japonica) from the F1 generation that 

were exposed to endosulfan. 

Table 35. Mean amount of time (s) from start of test until first mount attempt per 

trial by male Japanese quail (Coturnix japonica) from the F1 generation exposed 

to endosulfan. 

Dose

Average number

of Mounts

95% β Confidence

Interval* P‐value Average number

of Mounts

95% β Confidence

Interval* P‐value Average number

of Mounts

95% β Confidence

Interval* P‐value

0 2.45 2.80 2.20

30 1.90 .51  ‐  1.19 0.24 2.20 .53 ‐ 1.17 0.23 2.21 .66 ‐ 1.53 0.98

60 1.95 .52  ‐  1.21 0.29 2.70 .66 ‐ 1.40 0.85 2.75 .84 ‐ 1.86 0.27

125 2.00 .54  ‐  1.24 0.34 1.95 .46 ‐ 1.05 0.08 2.50 .76 ‐ 1.70 0.54

250 2.05 .55  ‐  1.27 0.40 2.32 .56 ‐ 1.23 0.35 2.65 .79 ‐ 1.82 0.38

* Confidence Intervals reflect comparison to control group (ppm = 0).  Ppm levels with a confidence interval not including 1.00 is considered to be significantly different

Trial 1 Trial 2 Trial 3

Omnibus Likelihood ratio p ‐ value = .773 Omnibus Likelihood ratio p ‐ value = .381 Omnibus Likelihood ratio p ‐ value = .740

F1Generation  ‐  Number of Mounts

Dose

0 59.4 14.8 85% 44.8 15.8 80% 37.9 15.2 85%

30 43.8 13.2 85% 27.2 11.8 90% 21.6 11.8 90%

60 60.1 13.2 90% 37.8 12.5 90% 10.6 2.9 100%

125 67.9 15.9 75% 37.4 12.0 95% 40.5 13.9 90%

250 44.3 11.9 90% 20.0 9.2 95% 15.8 9.5 94%

Mounted

Pairs

Average time to

first mount

Standard

Error

Mounted

Pairs

Average time to

first mount

Standard

Error

Mounted

Pairs

Average time to

first mount

Standard

Error

F1Generation ‐ Time to first Mount

Trial 1 Trial 2 Trial 3

Log rank p‐value = .795 Log rank p‐value = .790 Log rank p‐value = .444
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 

2008-March 2010 

27 

Table 36. Mean number of successful copulations within a 3 minute period per 

trial completed by Japanese quail (Coturnix japonica) from the F1 generation that 

were exposed to endosulfan. 

Table 37. Mean amount of time (s) from start of test until first successful 

copulation per trial by male Japanese quail (Coturnix japonica) from the F1 

generation exposed to endosulfan. 

No differences in number of mount attempts made within the three minute time period for each of the 

three trials for the copulatory behavior tests were observed for the F2 generation (Table 38). Birds from 

the 30 ppm treatment group in the F2 generation initiated their mount attempts earlier than any other 

dose group for all three trials, while control males took the longest (Table 39). Males from the 30 ppm 

treatment level also had a larger number of successful copulations than controls in the first trial (Table 

40), and took the least amount of time to achieve a successful copulation in the first and third trials (Table 

41). 

Trial 1

Dose

Average number

of Copulations

95% β Confidence

Interval* P‐value Average number

of Copulations

95% β Confidence

Interval* P‐value Average number

of Copulations

95% β Confidence

Interval* P‐value

0 0.35 0.50 0.75

30 0.60 .68 ‐ 4.35 0.26 0.55 .47 ‐ 2.59 0.83 1.00 .68 ‐ 2.62 0.41

60 0.60 .68 ‐ 4.35 0.26 0.50 .42 ‐ 2.40 1.00 0.75 .49 ‐ 2.05 1.00

125 0.35 .35 ‐ 2.85 1.00 0.45 .37 ‐ 2.22 0.82 0.40 .23 ‐ 1.26 0.15

250 0.40 .41 ‐ 3.12 0.80 0.71 .61 ‐ 3.27 0.42 0.53 .31 ‐ 1.61 0.41

* Confidence Intervals reflect comparison to control group (ppm = 0).  Ppm levels with a confidence interval not including 1.00 is considered to be significantly different

Trial 3

Omnibus Likelihood ratio p‐value = .193

Trial 2

Omnibus Likelihood ratio p‐value = .581 Omnibus Likelihood ratio p‐value = .873

F1Generation ‐ Number of Copulations

Dose

0 142.7 13.5 30% 114.4 18.0 40% 91.2 17.8 60%

30 105.6 16.7 55% 113.4 16.0 50% 58.9 14.4 85%

60 110.2 14.7 55% 128.4 15.8 40% 100.5 16.9 60%

125 143.8 14.3 30% 117.6 17.2 40% 130.8 16.1 35%

250 135.3 15.2 35% 91.9 20.2 53% 123.3 18.2 39%

Average time

to first

copulation

Standard

Error

Copulated

pairs

Average time

to first

copulation

Standard

Error

Copulated

pairs

Average time

to first

copulation

Standard

Error

Copulated

pairs

F1 Generation ‐ Time to first Copulation

Log rank p‐value = .212 Log rank p‐value = .755 Log rank p‐value = .013

Trial 1 Trial 2 Trial 3
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 

2008-March 2010 

28 

Table 38. Mean number of mount attempts initiated within a 3 minute time period 

per trial by male Japanese quail (Coturnix japonica) from the F2 generation that 

were exposed to endosulfan. 

Table 39. Mean amount of time (s) from start of test until first mount attempt per 

trial by male Japanese quail (Coturnix japonica) from the F2 generation exposed 

to endosulfan. 

Table 40. Mean number of successful copulations within a 3 minute period per 

trial completed by Japanese quail (Coturnix japonica) from the F2 generation that 

were exposed to endosulfan. 

Trial 2

Dose

Average number

of Mounts

95% β Confidence

Interval* P‐value Average number

of Mounts

95% β Confidence

Interval* P‐value Average number

of Mounts

95% β Confidence

Interval* P‐value

0 1.95 1.90 1.65

30 2.25 .75 ‐ 1.77 0.51 2.55 .88 ‐ 2.04 0.17 2.21 .85 ‐ 2.11 0.21

60 1.95 .64 ‐ 1.56 1.00 1.50 .49 ‐ 1.27 0.33 2.10 .81 ‐ 2.01 0.30

125 2.20 .73 ‐ 1.74 0.58 2.05 .69 ‐ 1.68 0.74 1.80 .68 ‐ 1.75 0.72

250 2.05 .68 ‐ 1.63 0.82 2.10 .71 ‐ 1.71 0.66 2.15 .83 ‐ 2.05 0.25

* Confidence Intervals reflect comparison to control group (ppm = 0).  Ppm levels with a confidence interval not including 1.00 is considered to be significantly different

F2Generation ‐ Number of Mounts

Trial 1

Omnibus Likelihood ratio p‐value = .945

Trial 3

Omnibus Likelihood ratio p‐value = .221 Omnibus Likelihood ratio p‐value = .659

Dose

0 84.9 18.2 60% 62.6 16.4 80% 47.9 15.3 80%

30 17.6 4.5 100% 3.4 0.5 100% 5.1 2.7 100%

60 57.7 16.9 80% 49.4 15.8 80% 22.8 9.3 95%

125 49.8 13.7 85% 26.9 11.1 95% 17.4 8.9 95%

250 50.2 13.7 85% 24.7 10.2 95% 17.1 9.0 95%

Mounted

Pairs

Average time to

first mount

Standard

Error

Mounted

Pairs

Average time to

first mount

Standard

Error

Mounted

Pairs

Average time to

first mount

Standard

Error

F2 Generation ‐ Time to first Mount

Trial 1 Trial 2 Trial 3

Log rank p‐value = .032 Log rank p‐value = .000 Log rank p‐value = .016

Trial 1 Trial 2

Dose

Average number

of Copulations

95% β Confidence

Interval* P‐value Average number

of Copulations

95% β Confidence

Interval* P‐value Average number

of Copulations

95% β Confidence

Interval* P‐value

0 0.35 0.60 0.65

30 1.10 1.34 ‐ 7.36 0.01 0.75 .59 ‐ 2.67 0.57 1.16 .90 ‐ 3.54 0.10

60 0.50 .54 ‐ 3.75 0.47 0.55 .40 ‐ 2.08 0.84 0.65 .46 ‐ 2.16 1.00

125 0.50 .54 ‐ 3.75 0.47 0.60 .45 ‐ 2.23 1.00 0.60 .42 ‐ 2.02 0.84

250 0.65 .74 ‐ 4.66 0.18 0.65 .49 ‐ 2.37 0.84 0.90 .68 ‐ 2.83 0.37

* Confidence Intervals reflect comparison to control group (ppm = 0).  Ppm levels with a confidence interval not including 1.00 is considered to be significantly different

F2Generation ‐ Number of Copulations

Omnibus Likelihood ratio p‐value = .043 Omnibus Likelihood ratio p‐value = .950

Trial 3

Omnibus Likelihood ratio p‐value = .268
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 

2008-March 2010 

29 

Table 41. Mean amount of time (s) from start of test until first successful 

copulation per trial by male Japanese quail (Coturnix japonica) from the F2 

generation exposed to endosulfan. 

 Organ Weights. No significant differences were observed with adult brain,kidney, liver, or 

thyroid weights for females from the F0 generation (Table 42), however kidney wieghts from adult 

males of the 250 ppm dose group were significantly smaller than those from the control and 30 ppm 

dose groups (Table 43). Additionally, male thyroid glands of the 125 and 250 ppm dose groups 

weighed less than those from controls. 

Table 42. Mean brain, kidney, liver, and thyroid gland weights (g) with standard 

error of deviation from adult female Japanese quail (Coturnix japonica) of the F0 

generation that were exposed to endosulfan (ppm). 

Dose

0 150.3 13.4 20% 112.8 17.2 50% 104.5 17.6 50%

30 72.8 14.4 80% 83.6 17.7 60% 44.5 13.6 85%

60 114.1 16.8 50% 111.8 17.7 45% 113.1 17.0 45%

125 118.2 16.6 45% 96.7 17.3 55% 96.7 16.0 60%

250 113.3 15.2 50% 105.7 16.1 60% 77.9 15.3 80%

Average time

to first

copulation

Standard

Error

Copulated

pairs

Average time

to first

copulation

Standard

Error

Copulated

pairs

Average time

to first

copulation

Standard

Error

Copulated

pairs

F2 Generation ‐ Time to first Copulation

Log rank p‐value = .005 Log rank p‐value = .863 Log rank p‐value = .005

Trial 1 Trial 2 Trial 3

Dose Average Std. Error Average Std. Error Average Std. Error Average Std. Error Average Std. Error

0 127.6 7.18 0.73 0.030 1.31 0.110 3.42 0.221 0.014 0.002

30 144.8 2.24 0.76 0.018 1.37 0.069 4.21 0.179 0.014 0.002

60 136.4 5.40 0.75 0.006 1.23 0.093 3.99 0.272 0.016 0.004

125 135.4 2.59 0.78 0.015 1.24 0.105 3.44 0.313 0.017 0.003

250 141.9 3.47 0.77 0.010 1.21 0.041 3.78 0.161 0.016 0.002

F0Generation Female Organ Weights

Body Weight Brain Kidneys Liver Thyroid

P‐value = .083* P‐value = .344 P‐value = .631 P‐value = .082 P‐value = .831

* P‐value calculated from Kruskall‐wallis test because homogenei ty of variance assumption violated in parametric ANOVA test.
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 

2008-March 2010 

30 

Table 43. Mean brain, kidney, liver, and thyroid gland weights (g) with standard 

error of deviation from adult male Japanese quail (Coturnix japonica) of the F0 

generation that were exposed to endosulfan (ppm). 

Females from the 125 ppm treatment level had larger brains than those from controls (Table 44). 

Although the p-value for male thyroid weights from the F1 generation was less than 0.05, post-hoc tests 

revealed that the differences between dose groups were not significant, due to the adjustment of multiple 

testing (Table 45). Cloacal gland size from F1 generation males approached significance (p=0.054), with 

glands from the 250 ppm group being approximately 13% larger than controls. 

Table 44. Mean brain, kidney, liver, and thyroid gland weights (g) with standard 

error of deviation from adult female Japanese quail (Coturnix japonica) of the F1 

generation that were exposed to endosulfan (ppm). 

Dose Average Std. Error Average Std. Error Average Std. Error Average Std. Error Average Std. Error

0 112.3 2.35 0.78 0.015 0.79 0.044 1.73 0.070 0.020 0.003

30 114.2 3.16 0.77 0.018 0.79 0.036 1.91 0.089 0.013 0.001

60† ‐ ‐ ‐ ‐ ‐ ‐ ‐ ‐ ‐ ‐

125 108.3 1.57 0.75 0.014 0.61 0.058 1.67 0.047 0.011 0.001

250 110.3 1.57 0.79 0.019 0.57 0.057 1.80 0.035 0.012 0.001

** Tukey post‐hoc results indicate that dose 0 has a significantly greater thyroid weight than dose 125 and dose

250.

† Only 1 observaƟon for dose 60.  With only 1 observaƟon, standard deviaƟon/error cannot be calculated and

therefore cannot be included in ANOVA test.

P‐value = .226 P‐value = .435 P‐value = .012* P‐value = .057 P‐value = .004**

* Tukey post‐hoc results indicate that dose 0 and dose 30 have significantly greater kidney weight than dose 250 at the 95%

confidence level.

F0 Generation Male Organ Weights

Body Weight Brain Kidneys Liver Thyroid

Dose Average Std. Error Average Std. Error Average Std. Error Average Std. Error Average Std. Error

0 137.0 4.24 0.73 0.009 1.39 0.003 4.10 0.206 0.014 0.004

30 142.2 2.09 0.76 0.013 1.28 0.072 4.10 0.243 0.011 0.001

60 129.7 2.64 0.75 0.006 1.30 0.058 3.69 0.226 0.008 0.001

125 144.7 1.32 0.78 0.010 1.32 0.027 4.16 0.241 0.012 0.002

250 144.4 3.21 0.75 0.030 1.30 0.026 4.10 0.281 0.016 0.003

† Kruskall‐wallis test indicates that dose 125 has a significantly greater brain weight averages compared to the control group

at 95% confidence level

F1Generation Female Organ Weights

Body Weight Brain Kidneys Liver Thyroid

P‐value = .002° P‐value = .038*† P‐value = .313* P‐value = .641 P‐value = .103*

* P‐value calculated from Kruskall‐wallis test because homogeneity of variance assumption violated in parametric ANOVA test.

° Tukey post‐hoc results indicate that dose 30, 250 and 125 have signi ficantly greater body weight average compared to dose

60 at 95% confidence level.
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 

2008-March 2010 

31 

Table 45. Mean brain, kidney, liver, and thyroid gland weights (g) with standard 

error of deviation from adult male Japanese quail (Coturnix japonica) of the F1 

generation that were exposed to endosulfan (ppm). 

No differences in brain, kidney, liver, or thyroid gland weights, or clocal gland size were observed in 

Japanese quail from the F2 generation (Tables 46 and 47). 

Table 46. Mean brain, kidney, liver, and thyroid gland weights (g) with standard 

error of deviation from adult female Japanese quail (Coturnix japonica) of the F2 

generation that were exposed to endosulfan (ppm). 

Table 47. Mean brain, kidney, liver, and thyroid gland weights (g) with standard 

error of deviation from adult male Japanese quail (Coturnix japonica) of the F2 

generation that were exposed to endosulfan (ppm). 

Dose Average Std. Error Average Std. Error Average Std. Error Average Std. Error Average Std. Error Average Std. Error

0 109.0 3.12 0.75 0.020 0.73 0.039 1.76 0.125 0.008 0.002 210.4 18.66

30 113.4 1.92 0.72 0.036 0.76 0.030 1.94 0.113 0.017 0.003 179.4 13.50

60 114.6 2.88 0.76 0.016 0.77 0.030 1.88 0.074 0.010 0.002 191.7 10.10

125 113.1 2.40 0.78 0.023 0.76 0.027 1.86 0.103 0.015 0.003 196.6 14.78

250 112.0 2.35 0.76 0.014 0.76 0.019 1.83 0.071 0.008 0.001 239.1 9.92

P‐value = .608 P‐value = .572 P‐value = .912 P‐value = .767 P‐value = .033 P‐value = .054

F1 Generation Male Organ Weights

Body Weight Brain Kidneys Liver Thyroid Coacal Gland

Dose Average Std. Error Average Std. Error Average Std. Error Average Std. Error Average Std. Error

0 138.1 2.72 0.77 0.009 1.19 0.035 4.02 0.192 0.014 0.0018

30 139.1 2.64 0.75 0.010 1.27 0.049 4.14 0.066 0.014 0.0010

60 132.8 1.53 0.75 0.013 1.33 0.035 4.43 0.367 0.011 0.0011

125 138.3 2.87 0.77 0.014 1.28 0.062 4.25 0.228 0.012 0.0019

250 135.8 1.81 0.77 0.011 1.22 0.053 3.98 0.377 0.015 0.0017

F2Generation Female Organ Weights

Body Weight Brain Kidneys Liver Thyroid

P‐value = .403* P‐value = .248 P‐value = .303 P‐value = .880* P‐value = .597

* P‐value calculated from Kruskall‐wallis test because homogeneity of variance assumption violated in parametric ANOVA test.

Dose Average Std. Error Average Std. Error Average Std. Error Average Std. Error Average Std. Error Average Std. Error

0 108.1 1.49 0.77 0.011 0.72 0.015 1.66 0.055 0.012 0.0080 221.1 14.23

30 111.1 2.41 0.76 0.015 0.77 0.018 1.79 0.053 0.011 0.0083 196.1 13.67

60 116.7 1.94 0.78 0.012 0.80 0.030 1.85 0.067 0.012 0.0086 201.3 6.43

125 117.4 1.79 0.79 0.017 0.79 0.025 1.92 0.072 0.010 0.0064 222.6 7.22

250 110.9 2.39 0.76 0.012 0.77 0.042 1.88 0.109 0.011 0.0074 201.4 16.48

* Tukey post‐hoc test indicate that dose 60 and dose 125 have signi ficantly greater body weight compared to dose 0 at a 95% confidence level.

P‐value = .009* P‐value = .322 P‐value = .345 P‐value = .157 P‐value = .894 P‐value = .432

F2Generation Male Organ Weights

Body Weight Brain Kidneys Liver Thyroid Coacal Gland
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 

2008-March 2010 

32 

Ovarian weights did not differ among treatment levels in the F0 and F2 generations, however ovaries 

from F1 females from the 60 ppm dose group were smaller than those from all other dose groups (Table 

48). Testes weight did not differ among dose groups in any generation. Although the number of 

dominant follicles per ovary did not differ among dose groups for any generation, follicle counts were 

lower in birds of the 60 ppm treatment level in the F1 generation compared to the other dose groups 

(Table 49). 

Table 48. Mean gonad weights with standard errors of the mean from adult 

Japanese quail (Coturnix japonica) exposed to endosulfan. 

Table 49. Mean ovarian follicle counts with standard errors of the mean from 

female Japanese quail (Coturnix japonica) that were exposed to endosulfan. 

   Ovarian Follicle Counts         

        F0 Generation       F1 Generation       F2 Generation

Dose           p = 0.155            p = 0.057            p = 0.273

(ppm) Average Std. Error Average Std. Error Average Std. Error

0 2.56 0.58 4.78 0.28 4.10 0.31

30 4.00 0.14 4.60 0.34 3.90 0.38

60 3.25 0.77 3.20 0.57 3.40 0.53

125 3.46 0.16 4.77 0.15 3.60 0.34

250 3.60 0.27 4.14 0.26 6.55 0.43

 Perivitelline Layer Sperm Penetration. No differences were observed with the number of 

hydrolysis points made by spermatozoa around the germinal disc for either the F0 or the F1 

generation (Table 50). 

                                  F0Generation                             F1Generation                           F2Generation

       Ovary Weight      Testes Weight       Ovary Weight      Testes Weight      Ovary Weight      Testes Weight

Dose                p = 0.407           p = 0.560                p = 0.003*           p = 0.750               p = 0.360           p = 0.355

(ppm) Average Std. Error Average Std. Error Average Std. Error Average Std. Error Average Std. Error Average Std. Error

0 3.27 0.79 3.72 0.29 5.56 0.31 3.34 0.09 2.85 0.64 3.4 0.15

30 4.95 0.45 3.10 0.23 5.02 0.41 2.92 0.35 3.80 0.55 3.32 0.20

60 4.32 0.85 3.14 0.17 3.09 0.63 2.97 0.34 2.85 0.78 3.66 0.11

125 4.49 0.27 3.19 0.22 5.00 0.36 3.08 0.16 3.26 0.57 3.28 0.15

250 5.13 0.31 3.18 0.22 4.91 0.35 1.07 0.34 1.98 0.53 3.13 0.25

*Mean ovarian weights from 60 ppm dpse group weigh less than those from all other treatment levels.
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 

2008-March 2010 

33 

Table 50. Number of hydrolysis points around the germinal disc made by 

spermatozoa from Japanese quail (Coturnix japonica) from the F0 and F1 

generations of quail that were exposed to endosulfan (ppm). 

           F1 Generation          F2 Generation

           # Sperm Holes          # Sperm Holes

          p‐value = 0.202         p‐value = 0.135

Dose (ppm) Average Std. Error Average

Std.

Error

0 26.75 5.53 10.00 2.47

30 14.13 2.18 22.85 5.55

60 20.34 3.61 17.19 3.61

125 18.55 2.57 25.99 6.67

250 17.43 4.27 19.89 2.46

 Fertility and Hatchability. Endosulfan did no apparent effects on fertility or hatchability in F1 and 

F2 chicks (Tables 51 and 52). 

Table 51. Percent fertility, hatchability, deaths occurring at approximately 1 and 2 

weeks of development, and F1 chicks that initiated but failed to hatch (pipped) in 

eggs produced by the F0 generation (exposure to endosulfan through feed only) 

of Japanese quail (Coturnix japonica). 

endosulfan (ppm) % fertility % hatchability % died wk 1 % died wk 2 % pipped

0 84.9 79 9.7 8 3.2

30 94.8 86.5 6.1 4.3 3.1

60 91.9 84.6 2.9 11.8 0.7

125 86.6 94.6 0 4 1.3

250 83.3 88.8 4 7.2 0
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 

2008-March 2010 

34 

Table 52. Percent fertility, hatchability, deaths occurring at approximately 1 and 2 

weeks of development, and F2 chicks that initiated but failed to hatch (pipped) in 

eggs produced by the F1 generation (exposure to endosulfan in ovo and through 

feed) of Japanese quail (Coturnix japonica). 

endosulfan (ppm) % fertility % hatchability % died wk 1 % died wk 2 % pipped

0 99.2 89.8 4.7 2.4 3.1

30 99.2 91.1 3.2 2.4 3.2

60 93.6 94.2 2.9 1.9 1

125 98.2 91.1 7.1 1.8 0

250 96.2 95.3 0 3.9 0.8

 Hormone Analysis. Endosulfan did not alter adult glandular T3 and T4 in any of the 

three generations (Tables 53-55). Adult serum estradiol and testosterone was likewise 

unchanged in the F0 and F1 generations. In the F2 generation serum estradiol was 

lower in females from the 250 ppm dose group than the 60 ppm group, and serum 

testosterone was lower in males from the 125 and 250 ppm dose groups than control 

males. 

Table 53. Average glandular thyroxine (T4) and triiodothyronine (T3) and serum 

estradiol and testosterone collected from F0 generation (exposure through feed 

only) adult Japanese quail (Coturnix japonica). 

Gender Dose Group

(ppm)

T3 (ng/ml) T4 (ng/ml)* Estradiol (pg/ml) Testosterone (pg/ml)*

N Mean SE N Mean SE N Mean SE N Mean SE

Female

Control 5 1.62 0.20 5 2.54 0.73 5 43.45 6.56 5 522.5 345.9

30 5 1.37 0.09 4 2.32 0.74 5 46.25 3.19 5 868.2 177.7

60 5 1.20 0.21 4 6.98 1.81 5 34.51 2.61 5 339.0 85.1

125 5 1.35 0.22 4 3.74 1.26 5 35.66 7.33 5 453.3 267.9

250 5 1.25 0.10 4 2.86 1.39 5 45.59 7.47 5 467.3 274.3

Male

Control 5 1.46 0.11 5 2.49 0.68 5 10.89 4.02 5 1986.3 732.4

30 5 1.50 0.09 4 2.50 0.50 5 14.55 8.85 5 791.8 136.2

60 5 1.60 0.12 5 2.34 1.07 5 10.86 2.89 5 909.6 126.1

125 5 1.58 0.14 5 2.71 0.51 5 9.88 1.37 5 1264.3 363.3

250 5 1.50 0.09 5 3.26 0.92 5 6.90 2.42 5 1157.0 297.8

P‐Value Dose = .820 Dose = .568 Dose = .563 Dose = .792
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 

2008-March 2010 

35 

Gender = .070 Gender = .235 Gender = .000       

(Females > Males)

Gender = .000         

(Males > Females)

* Analyzed using Log‐transformed data due to violations in ANOVA assumptions.      

Table 54. Average glandular thyroxine (T4) and triiodothyronine (T3) and serum 

estradiol and testosterone collected from F1 generation (exposure through 

embryonic exposure and feed) adult Japanese quail (Coturnix japonica). 

Gender

Dose

Group

(ppm)

T3 (ng/ml) T4 (ng/ml)* Estradiol (pg/ml) Testosterone (pg/ml)

N Mean SE N Mean SE N Mean SE N Mean SE

Female

Control 5 1.39 0.20 4 3.11 0.70 5 30.88 8.40 5 301.8 174.3

30 5 1.99 0.27 4 1.99 0.53 5 54.52 19.76 5 551.6 98.1

60 5 1.89 0.31 5 1.78 0.41 5 57.93 8.33 5 299.2 64.0

125 5 1.24 0.19 4 3.20 1.39 5 44.24 9.25 5 583.4 280.3

250 6 1.40 0.16 3 3.24 2.11 6 46.79 3.65 6 450.8 144.1

Male

Control 5 1.60 0.16 5 2.51 0.43 5 10.16 2.24 5 925.8 405.1

30 4 1.65 0.12 3 1.52 0.39 5 9.31 2.22 5 1249.1 437.0

60 5 1.63 0.15 5 2.80 0.58 5 9.23 0.62 5 2114.3 720.8

125 5 1.66 0.16 5 3.21 0.38 5 9.49 1.08 5 3280.4 1241.9

250 4 1.39 0.16 3 2.92 0.92 4 12.88 7.16 4 1693.4 586.0

P‐Value

Dose = .148 Dose = .459 Dose = .546 Dose = .159

Gender = .981 Gender = .607 Gender = .000       

(Females > Males)

Gender = .000         

(Males > Females)

* Analyzed using Log‐transformed data due to violations in ANOVA assumptions.      

Table 55. Average glandular thyroxine (T4) and triiodothyronine (T3) and serum 

estradiol and testosterone collected from F2 generation (embryonic exposure 

only) adult Japanese quail (Coturnix japonica). 

Gender

Dose

Group

(ppm)

T3 (ng/ml) T4 (ng/ml)* Estradiol (pg/ml) Testosterone (pg/ml)

N Mean SE N Mean SE N Mean SE N Mean SE

Female

Control 5 1.84 0.34 2 2.99 1.57 5 30.12 2.75 5 235.3 72.2

30 5 1.42 0.13 4 1.84 0.71 5 39.30 5.30 5 297.5 45.7

60 4 1.18 0.17 3 2.79 1.66 5 49.86 4.67 5 620.5 229.5

125 3 0.89 0.31 3 4.20 2.46 5 36.27 11.38 5 334.6 214.1
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 

2008-March 2010 

36 

250 5 1.38 0.34 4 3.89 1.45 5 18.83 4.98 5 511.3 279.6

Male

Control 5 1.66 0.09 4 2.69 0.09 5 10.17 1.86 5 2191.4 197.0

30 5 1.66 0.32 5 2.96 0.94 5 6.95 1.53 5 1759.1 425.1

60 5 1.80 0.14 5 1.72 0.24 5 5.61 1.96 5 1143.9 300.1

125 4 1.54 0.20 5 1.95 0.82 5 7.65 1.47 5 899.7 213.8

250 5 1.80 0.25 5 3.39 0.92 5 6.42 2.33 5 785.7 252.6

P‐Value

Dose = .388 Dose = .576

Dose * Gender =

.022

Dose * Gender =

.006 Gender  = .033         

(Males > Females) Gender = .712

* Analyzed using Log‐transformed data due to violations in ANOVA assumptions.      

8 Discussion 

Although endosulfan is sometimes described as being moderately to highly toxic to birds (Watts, 

2009) and is characterized as an endocrine disruptor that causes reproductive and developmental 

damage in humans and other mammals (Varayoud et al., 2008), it has often been demonstrated as 

a relatively weak endocrine disruptor in avian species. The adult Japanese quail acute oral LD-50 

for endosulfan is reported as 46 mg/kg, and the acute dietary LD-50 ranges from 1197-2906 mg/kg 

(Prakash et al., 2009; Hill and Camardese, 1986). For in ovo exposures, the chicken (white 

leghorn) embryo LD-50 ranged from 5.35 mg endosulfan/egg for a one-time injection at embryonic 

day 3 (Pourmirza, 2000) to 28 mg Endocel (35% endosulfan)/brown leghorn egg injected after 24h 

incubation (Mobarak and Al-Asmari, 2011). Mobarak and Al-Asmari (2011) stated 14 mg/egg to be 

a teratogenic dose level for brown leghorn chicks while 21 mg/egg was considered to be more of an 

embryotoxic dose. Chicks from their lowest dose of 7 mg/egg also had significantly more 

deformities compared to control chicks. In addition to skeletal and cartilaginous deformities, 

generalized edema, and hematomas were also reported at this lowest dose level. Whereas other 

studies have reported neurotoxic effects of endosulfan in birds as evidenced by ataxia, tremors, 

atypical posturing (Prakash et al., 2009; Hudson et al., 1984), limb paralysis (Mobarak and AlAsmari, 2011), depression and excitation (Qamar et al., 2012), no such effects were observed in 

the current study. 

As a potential endocrine disrupting chemical, endosulfan has been described as estrogenic and 

antiandrogenic, having been shown to cause chronic depression of circulating testosterone 

(Varayoud et al., 2008; Grunfield and Bonefeld-Jorgensen, 2004; Chatterjee et al., 2008). The 

estrogen-mimicking potential is somewhat limited, having been measured to be about six orders of 

magnitude lower than that of estradiol as measured by the E-screen test (Soto et al., 1994). 

Endosulfan failed to induce a uterotrophic response in ovariectomized rats but induced changes in 

progesterone receptor and estrogen receptor alpha expression (Varayoud et al., 2008). The first 

study to assess endosulfan’s potential for reproductive toxicity in birds (Japanese quail) found no 
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 

2008-March 2010 

37 

effects on egg laying, eggshell thickness, or hatchability from birds that were fed feed treated with 

Endocel (35% endosulfan) at levels as high as 1000 ppm for 20 weeks (Prakash et al., 2009). Male 

Japanese quail that received 5 mg/kg endosulfan in feed had reduced testes relative weights and 

leukocyte counts compared to controls, while birds exposed to 25 and 50 mg/kg Thiodan EC 35 

(35% endosulfan) had suppressed onset of puberty (as measured by appearance of foam in feces) 

and a delay in timing of mounting pen mates (Qamar et al., 2012). No alterations in serum 

testosterone levels were observed. 

Although the information concerning endosulfan toxicity in an avian species gained from this study 

may be useful given the dearth of information available for avian species, the main objective of this 

study was to assess the value added by use of the F2 generation compared to a study that would 

test no further than the F1 generation. The current guideline for the avian two-generation toxicity 

test that has been proposed by the US EPA (Touart 2005) is a large, long-term experiment that is 

arguably resource expensive. The current study makes use of a large number of animals. In order 

to ensure a sufficient sample size for adequate statistical analysis and the production of 

subsequent generations for all dose levels, extra birds were produced than directly needed for 

inclusion in the groups that were measured and assessed for toxicity. The current trend in 

toxicology is one of a reduction in animal use and an increase in the reliance on in vitro versus in 

vivo studies as applicable. It is acknowledged that the endpoints in this study require the exposure 

of a living system, and non-animal alternatives are not available to replicate the complex processes 

of reproduction and development in an intact organism. However, the primary objective of this 

study is the justification of the additional F2 generation. The current study took approximately 

thirteen months to complete all in-life portions, with many additional months required for histological 

and hormone analyses. Though recent refinements in the methodologies for this avian twogeneration test have reduced the amount of time required to conduct this test, the investigation was 

temporally as well as financially expensive. 

Assessment of the utility of the F2 generation was challenging using the mildly estrogenic model 

endocrine disrupting chemical endosulfan. Endosulfan induced minimal changes in physiology and 

behavior, with many of the effects not particularly dose dependent or consistent across 

generations. What was fairly consistent was the lack of effects seen in adult male and female 

specific measures in the F2 generation (Tables 53 and 54). While males at the lowest treatment 

level did display altered copulatory behaviors compared to controls, observed effects (earliest 

initiation to mount females, earliest time to successful copulation, and greatest number of 

successful copulations) were not considered adverse (Table 55). Additionally, although some 

changes in adult hormone levels were observed, no correlating adverse functional reproductive 

effects occurred. 

Table 56. Comparison of adult male Japanese quail (Coturnix japonica) measures 

between the F1 (exposed to endosulfan in ovo and through feed) and F2 (in ovo

exposure only) generations. ND = no difference. 
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 

2008-March 2010 

38 

Table 57. Comparison of adult female Japanese quail (Coturnix japonica) 

measures between the F1 (exposed to endosulfan in ovo and through feed) and 

F2 (in ovo exposure only) generations. ND = no difference. 

Table 58. Comparison of results of F1 and F2 generation male copulatory 

behavior tests in Japanese quail (Coturnix japonica) exposed to endosulfan. ND = 

no difference. 

Yamashita et al. (2011) evaluated changes in Japanese quail exposed to low doses of estradiol as 

a positive control to determine sensitive endpoints for use in a one-generation avian endocrine 

disrupting toxicity test. Adult quail were fed a diet containing 0.3, 3, and 30 ppm 17 β-estradiol for 

six weeks. Decreased body weight, testicular and ovarian atrophy, decreased spermatogenesis, 

decreased egg production and fertility rate, and a reduction in eggshell thickness were observed in 

birds receiving 30 ppm estradiol compared to controls. Though eggs produced by the F1 

generation of the current endosulfan study did have thinner shells than controls at the 125 and 250 

ppm dose levels, egg production was not reduced in either generation. In fact, egg production was 

increased for a short time in birds from the 250 ppm group. A stimulatory effect of estradiol on egg 

production has been documented previously (El-Ghalid, 2009), and a hormetic effect of estradiol 

(and xenoestrogens) on egg production is likely. The sperm parameters and fertility endpoints 

measured in this study were also unchanged by endosulfan in both generations. Histological 

Male Specific Measures F1 F2

male onset to maturity ND ND

sperm concentration ND ND

sperm motility ND ND

% progressive sperm 250 ppm more progressive than 0, 30, 60 ppm 0& 60 ppm more progressive than rest

perivitelline layer penetration ND ND

testes weight ND ND

cloacal gland area 250 ppm largest (p = 0.054) ND

Female Specific measures F1 F2

female onset to maturity 30 ppm sooner than 0 and 60 ppm ND

egg production 250 ppm highest lay rate btwn. 1st & 3rd week ND

eggshell thickness 125& 250 ppm less than controls ND

eggshell strength 250 ppm greater than controls ND

ovary weight 60 ppm smallest ND

dominant follicle count ND ND

Copulatory Behavior Measure F1 F2

time until 1st mount attempt ND 30 ppm earliest initiation in trial 3

time until 1st copulatory success 125 ppm longer than 30 ppm in trial 3 30 ppm earliest success in trials 1& 3

# attempts / 3min trial interval ND ND

# successful copulations ND 30 ppm more than controls in trial 1
Toxicology Study No. 87-XE-08JG-10, Protocol No. 08JG-80-08-04-02, September 

2008-March 2010 

39 

results (Appendix G) suggest that endosulfan caused no effects on male or female reproductive 

tissue as well. Yamashita et al. (2011) also reported increased incidences of persistent right 

oviducts in females caused by low doses of estradiol. Only one bird in the current study from the 

250 ppm dose group exhibited this abnormality, making the interpretation questionable and not 

likely associated with exposure to endosulfan. 

9 Conclusion 

There appears to be no clear benefit to produce a generation of Japanese quail beyond the F1 

generation for the assessment of low potency endocrine disrupting chemicals exemplified by 

endosulfan based on the observed effects that were caused by the exposure levels selected for this 

study. In this case, a one-generation or extended one-generation avian toxicity test may be more 

appropriate. It is likely that the results of the Endocrine Disrupting Screening Program‘s Tier 1 

toxicity tests could be used to help determine if a candidate chemical would be more appropriately 

screened by an (extended) one- or a full two-generation test. 

10 Point of Contact 

If you have questions concerning this study, contact Dr. Michael J. Quinn, Health Effects Research 

Program at 410-436-1064. 

 MICHAEL J. QUINN 

 Biologist 

 Health Effects Research Program 

APPROVED: 

MARK S. JOHNSON 

Program Manager 

Health Effects Research Program 
B-1 

APPENDIX A 

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Yamashita, R., A. Oshima, Y. Hasegawa-Baba, M. Wada, K. Shibuya. 2011. Endocrine 

disrupting effects of low dose 17 β-estradiol (E2) on the Japanese quail (Coturnix japonica) were 

detected by modified one-generation reproduction study. J. Toxicol. Sci., 36: 43-54. 
Toxicology Study No. 87-XE-09HW-08, Protocol No. 09HW-80-08-03-01, 

April–June 2008 

APPENDIX B 

QUALITY ASSURANCE STATEMENT 

For: DTOX Study No. 87-XE-09HW-08, Protocol No. 09HW-80-08-03-01, titled 

“Sensitivity Determination of the Humoral Response to Foreign Red Blood Cells in the 

Northern Bobwhite, April-June 2008.” The following critical phases were inspected/ 

audited by the USAPHC (Prov)-Quality Systems Office: 

PRE IN-LIFE PHASE OF THE STUDY

Critical Phase Inspected/Audited Date Inspected 

/Audited 

Date Reported 

to 

Management/SD 

GLP Protocol Review 

IN-LIFE PHASE OF THE STUDY

Critical Phase Inspected/Audited 

Date 

Inspected 

/Audited 

Date Reported to 

Management/SD 

Test System – Facilities 

Test System – Receipt 

Test System – Identification 

Test Article – Control 

Test Article – Dosing 

Maintenance and Calibration of Equipment 

Compliance with DTOX Protocols 

Compliance with DTOX SOPs 

Reagents and Solutions 

Clinical Testing – Bleeding Technique 

POST IN-LIFE PHASE OF THE STUDY

Critical Phase Inspected/Audited 

Date 

Inspected 

/Audited 

Date Reported to 

Management/SD 

Final Study Report Review 

Study Raw Data Review 

Note: All findings were made known to the Study Director and the Program Manager at the time 

of the audit/inspection. If there were no findings during the inspection, the inspection was 

reported to Management and the Study Director on the date shown in the table. 

_____________________________________ __________________ 

Michael P. Kefauver Date 

GLP Assessor, QSO 

_____________________________________ __________________ 

Gene Sinar Date 

Team Leader, QSO
APPENDIX C 

ARCHIVES AND STUDY PERSONNEL 

C-1. ARCHIVES 

 a. All raw data, documentation, records, protocol, and a copy of the final report 

generated as a result of this study will be archived in Bldg. E-2100, USAPHC (Prov), for 

a minimum of five (5) years following submission of a final report. 

 b. Records on animal receipt, diet, and facility environmental parameters will be 

archived by the Veterinary Medical Division, Toxicology Directorate, for a minimum of 

five (5) years following submission of the final report to the Sponsor. If the report is 

used to support a regulatory action, it shall, along with all supporting data, be retained 

indefinitely. 

 c. Archived SOPs and maintenance and calibration logbooks may be found in Room 

1026, Building E-2100, USAPHC (Prov), Aberdeen Proving Ground, Maryland 21010. 

 d. The present study uses USAPHC (Prov) Toxicology Study No. 87-XE-08JG-10, 

Protocol 08JG-80-08-04-02 for all filings. 

C-2. PERSONNEL. The following USAPHC (Prov) personnel participated in the 

studies reported herein. 

a. Directorate of Toxicology: Dr. Michael Quinn, Michelle Cook, M.S., Alicia Shifflet, 

A.S., Allison Jackovitz, B.S., Craig A. McFarland, D.V.M., Ph.D., Mark S. Johnson, 

Ph.D. 

 b. Quality Systems Office: Mr. Gene Sinar, Mr. Michael Kefauver 
D-1 

APPENDIX D 

ANIMAL USE PROTOCOL WITH MODIFICATIONS 

 
ANIMAL USE PROTOCOL 

TOXICOLOGY DIRECTORATE 

U.S. ARMY CENTER FOR HEALTH PROMOTION 

AND PREVENTIVE MEDICINE 

ABERDEEN PROVING GROUND, MD 21010-5403

PROTOCOL TITLE: Two-generation test for endocrine disruption and reproductive toxicity 

in Japanese quail (Corturnix japonica) using endosulfan. 

 

PROTOCOL NUMBER: 

PRINCIPAL INVESTIGATOR/STUDY DIRECTOR: 

Michael J. Quinn, Jr., Ph.D. 

Biologist 

Health Effects Research Program 

Toxicology Directorate (410) 436-1064 

U.S. Army Center for Health Promotion and Preventive Medicine 

Aberdeen Proving Ground, MD 21010-5403 

CO-INVESTIGATOR(S): 

Mark Johnson, Ph.D., DABT 

Biologist 

Health Effects Research Program 

Toxicology Directorate (410) 436-5081 

U.S. Army Center for Health Promotion and Preventive Medicine 

Aberdeen Proving Ground, MD 21010-5403 

Emily LaFiandra, Ph.D. 

Toxicologist 

Health Effects Research Program 

Toxicology Directorate (410) 436-7749 

U.S. Army Center for Health Promotion and Preventive Medicine 

Aberdeen Proving Ground, MD 21010-5403 

Craig McFarland, Ph.D. 

Biologist 

Health Effects Research Program 

Toxicology Directorate (410) 436-7784 

U.S. Army Center for Health Promotion and Preventive Medicine 

Aberdeen Proving Ground, MD 21010-5403 

Amanda Hess-Ruth 

Biologist 

Toxicity Evaluation Program 

Toxicity Directorate (410)436-3973 

U.S. Army Center for Health Promotion and Preventive Medicine 
Aberdeen Proving Ground, MD 21010-5403 

SPONSOR: 

Christiana Grim, D.V.M. 

Project Officer 

Office of Science Coordination and Policy 

U.S. Environmental Protection Agency 

1200 Pennsylvania Ave., NW (7203) 

Washington, DC 20460 

 Tel: 202-564-8458 

I. NON-TECHNICAL SYNOPSIS: 

 Many military chemicals are known or suspected to be endocrine active and have been 

found in soil and water on military installations at significant concentrations. Avian species 

make up a large percentage of the biodiversity in these areas and have great potential for 

exposure to these contaminants. The purpose of this study is to establish a new laboratory 

testing procedure that can be used to assess the impact of endocrine active chemicals upon avian 

health and reproduction. Data collected from such tests may be used in the assessment of risks to 

birds exposed to military environmental contaminants. These data from a two-generation test are 

principally intended to evaluate the adverse consequences of exposure to a “possible” endocrine 

disruptor to the reproductive viability of first generation birds, but may also provide information 

to confirm a chemical as an endocrine disruptor. 

II. BACKGROUND: 

II.1. Background: The U.S. Environmental Protection Agency (EPA) is implementing an 

Endocrine Disruptor Screening Program (EDSP) comprised of a battery of Tier 1 screening 

assays and Tier 2 tests. One of the Tier 2 tests under development is a two-generation avian 

assay using Japanese quail (Coturnix japonica). This test is designed to characterize doseresponse relationships and immediate and latent adverse consequences associated with exposure 

to putative endocrine modulating compounds. The avian two-generation assay serves to evaluate 

potential population effects of endocrine modulating compounds on myriad avian species, and 

represents terrestrial, oviparous wildlife species in general. This study will use endosulfan to 

fully evaluate the proposed test method for its ability to detect adverse effects and characterize 

dose responses in a two-generation paradigm, and to develop and optimize a protocol for the 

avian two-generation assay for characterizing endocrine disruptors. This work will help to 

determine the value added by examining endocrine endpoints over two generations in 

comparison to tests that only include one generation, such as the Ecological Effects Guidelines 

for avian reproduction tests (EPA, 1996). 

II.2. Literature Search for Duplication: 

II.2.1. Literature Source(s) Searched: 
 A number of databases were consulted in an effort to obtain information on the toxicity of 

endosulfan in the Japanese quail as well as a determination for duplication of effort. The 

databases checked were: 

 

 a. MEDLINE®: National Library of Medicine Database (1950 – 2008) 

b. RTECS: Registry of Toxic Effects of Chemical Substances (1960 – 2007) 

Science (1996-1999) 

c. TOXLINE® (1900 – 2006) 

d. PubMed (1980-2008) 

e. Adverse Reaction Database (2007-2008) 

f. Toxfile (1965-2008) 

g. Inside Conferences (1993-2008) 

h. Biological and Chemical Index (1983-2008) 

i. DTIC, STINET: Defense Technical Information Center Private Scientific and Technical 

Information Network (1980-2008) 

j. AGRICOLA: USDA National Agricultural Library (1970 – 2008) 

k. International Phamacology Abstracts (1970-2007) 

l. BIOSIS Toxline ® (1969-2008) 

m. Pharm-line ® (1978-2002) 

n. Pascal (1973-2008) 

o. Global Health (1983-2008) 

p. CBRN START: Chemical, Biological, Radiological and Nuclear. Scientific and 

Technical 

q. Analysis Research Tool (1980-2008) 

r. CAB Abstracts (1972-2008) 

s. EMBASE (1974-2008) 

t. NTIS: National Technical Information Service (1964-2008) 

u. BIOSIS Previews (1926-2008) 

II.2.2. Date of Search: 11 February 2008, by the research librarian at Wood Technical 

Library.

II.2.3. Period of Search: 1980-2008. 

 No date limits were applied to any of the databases. The time periods established for each of 

the individual databases were the defaults used, as shown above.

 

II.2.4. Key Words of Search: Japanese quail, Coturnix, quail, endosulfan, tiovel, thiotox, 

thiosulfan tionel, thiosulfan, thionex, thiodan, thimul, thifor, malix, insectophene, hildan, 

endosulfan, endocel, devisulphan, crisulfan, chlorthiepin, beosit, benzoepin, 115-29-7. 

II.2.5. Results of Search: Few studies were located using the above described search and an 

independent literature search. 

1. Acute Exposures. The 5-day dietary LC50 for endosulfan in 14-day old Japanese 

quail was found to be 1250 ppm and 2906 ppm (FWS, 1975; Hill and Camardese, 

1986). In mallards, the oral LD50s at 36 h, 7 d, 30 d, and 6 months of age were 27.8, 
6.47, 7.89, and 34.4 mg/kg body weight, respectively (Hudson et al., 1972). In 

pheasants, the LD50 values rage from 80 to >320 mg/kg (Hudson et al., 1984). 

2. Chronic Exposures. Broiler chicks exposed to 2 ppm endosulfan for 8 wk had a 

decrease in serum globulin and acetyl cholinesterase activity, total leukocyte and Tlymphocyte counts, and reductions in spleen and bursa of Fabricius weights (Garg et 

al., 2004a). Bursal follicle sizes and thymus lymphocyte numbers also decreased. 

Under the same conditions, broiler chicks also exhibited bone effects (Garg et al., 

2004b): the zones of proliferating, maturing, degenerating and calcifying cartilage 

cells were reduced in width, and a reduction in active osteons was observed. 

3. In Ovo Exposures. At 100 and 500 mg/kg, endosulfan in corn oil reduced the 

hatchability of white leghorn chickens by 24 and 53%, respectively, compared to 

controls (Dunachie and Fletcher, 1969). Previous studies had shown 5 mg/egg to 

reduce hatchability in white leghorns by up to 60% (Dunachie and Fletcher, 1966). 

 Spraying or dipping layer eggs with endosulfan in alcohol or acetone at 0.5 to 5% 

concentrations resulted in sterility of the gonads after the gonads were removed at 

days 8 and 9 of incubation and cultured on agar medium with 3 drops of a 0.5 to 1.0 

g/l endosulfan solution (Lutz and Lutz-Ostertag, 1972). 

4. In Vitro Studies. Alpha-endosulfan was observed to bind weakly to the chicken 

estrogen receptor (Matthews et al., 2000). 

III. OBJECTIVE/HYPOTHESIS: 

 The objective of this research is to determine the value added by examining endocrine 

endpoints in a two-generation avian reproductive toxicity test paradigm. 

IV. MILITARY RELEVANCE: 

Many chemicals used by the military that are known or suspected to be endocrine active have 

been found in environmental media at many military installations. Currently, few methods exist 

to ascertain the magnitude of these effects, although some have recently been proposed. Most 

species of birds are protected by the Migratory Bird Species Act and some are protected by the 

Endangered Species Act. As such, the DoD is by executive order obligated to abide by these 

laws. Presence of some species at specific installations (e.g. Bald Eagle at Aberdeen Proving 

Ground) has had a significant impact on reducing and curtailing military activity, therefore 

affecting readiness of our forces. Proactive collaboration in the development of methods that 

may, in the future, be mandated would serve the military, the Environmental Protection Agency, 

and the federal government at large. 

V. MATERIALS AND METHODS: 

V.1. Experimental Design and General Procedures: 

V.1.1. Experimental Design: 

 General Design: 

The general design of this study is illustrated in Figure 1. We will begin to expose the 
parental (P) generation at four weeks of age and throughout the rest of the birds’ lives. 

Eggs from this generation will be incubated and hatched to produce the first generation 

(F1). The F1 generation will be exposed to the treatments in ovo (embryonic) through 

maternal deposition and through feed throughout the entirety of their lives. Eggs from 

this generation will be incubated and hatched to produce the second generation (F2). The 

F2 generation will only be exposed to endosulfan in ovo through maternal deposition; all 

feed for the F2 generation will be untreated. Measures of reproductive behavior, 

circulating hormone levels, egg quality, reproduction, and development will be assessed 

in the P, F1, and F2 generations as described below. Additional measures of growth, 

reproductive maturity, immunocompetence, will be assessed in the F1 and F2 generations 

as described below. 

Figure 1. Timeline of major events in avian two-generation toxicity test. 

Chemical Treatments: 

The sample size for each treatment will be 16 to ensure that there are 12 breeding pairs in 

the parental generation control group at the end of the treatment period that have 

successfully produced eggs. The lowest reported LC50 from a five-day dietary exposure 

to endosulfan in 14-day old Japanese quail was found to be 1250 ppm (FWS, 1975). The 

treatment levels will be 0 (control), 30, 60, 125, and 250 ppm. The highest dose to be 

administered is one-fifth of the lowest known LC50 value and should not cause mortality 

or other severe signs of parental toxicity that will preclude the evaluation of reproductive 

parameters. The lowest concentration should produce no effects on adult health or 

reproductive measures. The test diets will be mixed and assayed at Wildlife International 

Ltd., Easton, MD, USA. Stability analyses and homogeneity and concentration 

verifications will be made every time Wildlife International Ltd. mixes new diet during 

the treatment period, according to GLP standards. 
Eggs: 

To generate the P generation, a minimum of 60 eggs will be hatched for each treatment 

level, to provide sufficient numbers of chicks to establish at least 12 breeding pairs (16 

pairs is the goal) per P test level. Sixteen males and females will be used for the study, 

beginning exposure to the treatments at week 4. Optimally, 6 eggs will be set for hatch 

for each parental pair of the P generation to ensure that a minimum of 60 eggs will be 

hatched for each test level, providing sufficient numbers of chicks to establish at least 12 

breeding pairs (16 is the goal) per F1 test level. If a treatment level produces insufficient 

eggs to set the minimum, that level will be dropped from further analysis in the test and 

eggs will not be set to hatch. Fertilized eggs not set to hatch will be frozen prior to being 

discarded. Optimally, 6 eggs will be set for each parental pair of the F1 generation, but a 

minimum of 40 eggs will be set and hatched for each treatment level. 

During the treatment period, all eggs, with the exception of those that are cracked, broken 

or abnormal, or used for eggshell or biochemical measurements, will be incubated. Only 

those eggs from the final week cohorts will be allowed to hatch. Eggs will be collected at 

least once daily, numbered according to pen of origin, and stored in a chilled 

environment; broken eggs will be numbered, recorded and then discarded. The total 

number of eggs laid per pen will be accounted for at the end of the study. 

Fertility and embryo viability will be determined by removing the eggs out of the 

incubator and candling them. Eggs are candled for fertility after 8 days of incubation and 

again for viability of embryos approximately 2 days before hatching (day 15 if the quail 

population at the laboratory generally hatches on day 17). Except for eggs incubated to 

hatch for production of subsequent generations, the late-stage embryos (day 15) will 

undergo necropsy and observations. During candling, eggs should not be allowed to cool 

to room temperature, since this may delay embryonic development. Those chicks that 

have not hatched within approximately 24 hours of the majority of chicks hatching will 

be considered unhatched. No assistance will be given to chicks during hatching, however 

chicks that remain alive but have no clear ability to hatch will be humanely removed 

from the egg and decapitated immediately. 

During the treatment periods, one egg per pen will be collected from odd numbered pens 

in odd numbered weeks and from even numbered pens during even numbered weeks. 

These eggs will be used to assess eggshell strength and eggshell thickness. Eggshell 

strength will be measured by an Egg Force Reader (Orka Technology Ltd.) that tests the 

breaking point of an egg’s shell. Eggshell thickness will be measured by an Egg Shell 

Thickness Gauge (Orka Technology Ltd.) that measures the thickness of egg shells 

through ultrasound or by micrometers or calipers. 

General Reproductive Fitness Measures and Observations: 

During the treatment period, the parental birds will be observed daily to detect any overt 

signs of toxicity or other clinical signs of illness or trauma. Observations will include 

mortality and clinical signs of toxicity such as lethargy, depression, wing droop, ruffled 

feathers, etc. Any injuries sustained and subsequent treatment will be recorded. If a pair 

is removed during the study due to death, injury, etc., data from all preceding weeks are 
evaluated. If data are used from a pen in which mortality occurred or a bird was injured, 

this will be explained. Clearly moribund individuals or those otherwise in severe distress 

will be euthanized, as described below. 

Prior to and during the treatment periods, the following information will be recorded in 

the study’s notebook or worksheets: 

 Eggs laid 

 Eggs cracked 

 Eggs broken 

 Description of all egg abnormalities 

 Eggs set 

 Eggshell strength 

 Eggshell thickness 

 Fertile eggs 

 Infertile eggs 

 Early embryonic deaths 

 Late embryonic deaths 

 Eggs that hatch 

 Eggs that do not hatch 

 Egg weight at 8 days and 24 hours prior to hatch 

The following clinical/fitness observations on all hatched offspring will be recorded 

during the treatment periods in the study’s notebook or worksheets: 

 Day of hatch 

 Hatchling mortality 

 Hatchling morphology and/or abnormalities 

 Clinical signs of toxicity 

 Number of 14-day-old surviving chicks 

 Chick body weight at hatching and 14 days after hatching 

 Sex of F1/F2 chicks 

All adult birds will undergo necropsy and gross pathology assessment. The wet weight of 

the brain, liver, kidneys, gonads, and thyroid glands will be recorded as soon as possible 

after death/euthanasia. Any morphologic abnormalities will be recorded in the study’s 

notebook or necropsy sheets. Adult birds that die, or are euthanized during the course of 

the treatment period, will be necropsied similarly. Histopathology will be performed on 

the brain, right thyroid gland, liver, bursa of Fabricius, kidneys, and gonads. The left 

thyroid gland will be stored for glandular thyroxine (T4) content. All late-stage embryos 

(2 days prior to hatch) that are not allowed to hatch will be decapitated and undergo 

necropsy as described above for adults. 

Birds (adults or chicks) which are in severe distress will be euthanized. If one member of 

a pair dies or is euthanized during the treatment period, the other member of the pair will 

be euthanized as well. 
Biochemical/Endocrine Measurements: 

Measurements of thyroid function will include thyroid gland weight, glandular T4 

content, and thyroid histology/histopathology. Measurements will be taken during 

embryonic and adult life stages. Eggs that are set, but not in the cohorts to be hatched, 

can be used for embryonic measurements. 

Blood from embryos removed from the shell that are used for necropsy and egg yolk 

samples shall be collected for steroid hormone (estradiol and testosterone) analysis. 

Blood collected from adults at the time of necropsy shall also have serum steroid analyses 

performed. 

Vitellogenin is a protein that is generally controlled by estrogen. It is present in females 

of oviparous species during egg development. It is not expressed in normal males due to 

lack of estrogen signal, however in males that have been exposed to estrogenic 

contaminants, it is expected that vitellogenesis will be stimulated. Vitellogenin will be 

measured from blood of the late embryos used for steroid hormone analysis, thyroid 

hormone analysis, and necropsy/pathology at CHPPM or a contracted lab. It will also be 

measured in adult blood at necropsy. 

Behavioral measures: 

Initiation of normal male sexual behavior is important for successful reproduction. In 

reproductively mature males of the F1 and F2 generations, preferably when at least 90% 

of control males have reached reproductive maturity, trials shall be performed to assess 

reproductive behavior including time to mount attempts, number of mount attempts, and 

number of successful cloacal contacts. Males and females will be paired in the F1

generation following completion of the behavioral measures. Birds from the F2

generation will be euthanized upon completion of this measure. 

Growth: 

Growth will be measured at least every three days in chicks up to at least week 2 of age. 

Growth measures will include body weight, tarsus length, and the length from the tip of 

the beak to the culmen. 

Immune Measures: 

The measurement of antibody production in response to foreign red blood cells is a 

common method for assessing humoral immunity. Quail will be immunized with chukar 

partridge (Alectoris chukar) erythrocytes (upon availability) because Japanese quail are 

minimally sensitive to sheep, human, chicken, turkey, and duck erythrocytes and do not 

produce immunoglobulins to human, bovine, and mouse albumins. The ability to 

perform this measure is dependent on the availability of chukar partridge red blood cells, 

since this material is normally only available right before and during hunting seasons. 

Adult quail to be challenged will receive 0.1 ml of a 1.0% suspension of chukar red blood 

cells (CRBCs) in saline injected into the jugular vein. Blood will be collected from the 

jugular vein 6 days after the CRBC injection, and serum may be frozen until antibody 

titers are determined. Total (IgM and IgG) and 2-mercaptoethanol-resistant (IgG) 
antibody activities will be measured by a hemagglutination assay at CHPPM or at a 

contracted lab. 

Cell mediated response will be measured in juveniles by using a 0.1 ml dose of 1 mg/ml 

phytohemagglutinin (PHA-P) in phosphate-buffered saline (PBS). One wing web will 

receive an injection with the PHA solution while the other wing web will receive an 

injection of PBS alone. The thickness of each wing web will be measured to the nearest 

0.01 mm immediately before and 24 hr after the injections using pressure-sensitive 

micrometers with a low-tension spring that does not crush the skin. A stimulation index 

will be calculated as the difference in the change in thickness of the PHA-injected wing 

web from the change in thickness of the PBS-injected wing web. 

This study will be conducted in compliance with Good Laboratory Practice (GLP) 

regulations found in the Toxic Substances Control Act (TSCA): 40 CFR (Code of 

Federal Regulations) 792, plus amendments. The investigators and technicians will 

adhere to The Guide for Care and Use of Laboratory Animals (U.S. Department of 

Health, Education, and Welfare Publication No. NIH 86-23, 1996). 

 

V.2. Data Analysis: All observational data will be recorded in the laboratory notebook and 

using LabCat® where applicable. The analysis of variance (ANOVA) assumptions will be 

checked, and if they are met, a one-way ANOVA will be applied followed by a multiple 

comparisons test to compare treatment groups. If data do not meet the ANOVA assumptions, 

data may be transformed appropriately or a nonparametric ANOVA may be applied. 

Observational categorical data may be tested with a Fisher’s Exact or Chi Square test. Statistical 

significance is defined as p < 0.05. 

V.3. Laboratory Animals Required and Justification: 

V.3.1. Non-animal Alternatives Considered: The objective addressed by this study is to 

determine the value added by examining endocrine endpoints in an avian reproductive toxicity 

test paradigm. Non-animal alternatives would not provide the necessary information.

V.3.2. Animal Model and Species Justification: The Japanese quail was chosen for this study 

because it is an avian species that is both often used in toxicity tests and breeds readily in a 

laboratory setting. In addition, these birds only take six weeks to reach sexual maturity, which 

greatly reduces the time required to produce multi-generations when compared to other similar 

species, such as the northern bobwhite which takes an average of six months to reach 

reproductive maturity. 

 

V.3.3. Laboratory Animals: 

V.3.3.1. Genus and Species: Coturnix japonica

V.3.3.2. Strain/Stock: In-bred; nonspecific 
V.3.3.3. Source/Vendor: University of Maryland, Department of Animal and Avian Sciences, 

College Park, MD. 

V.3.3.4. Age: Eggs - adult

V.3.3.5. Weight: Adult weight = ~150g 

V.3.3.6. Sex: Males and females

V.3.3.7. Special Considerations: None

V.3.4. Number of Animals Required (By Species): 

1) Japanese quail: 

a. P generation: Minimum of 60 eggs set to incubate for 5 treatments = potential 

for maximum of 300 chicks; 16 males and 16 females from each treatment 

(80 males and 80 females total) will be kept for exposure to endosulfan. 

b. F1 generation: Minimum of 60 eggs set to incubate for 5 treatments = 

potential for maximum of 300 chicks; 16 males and 16 females from each 

treatment (80 males and 80 females total) will be kept for exposure to 

endosulfan. 

c. F2 generation: Minimum of 40 eggs set to incubate for 5 treatments = 

potential for maximum of 200 chicks; minimum of 12 males and 12 females 

from each treatment (60 males and 60 females total) will be kept for behavior 

analyses. 

V.3.5. Refinement, Reduction, Replacement: 

V.3.5.1. Refinement: The exposure levels used in this study are based on data from previous 

studies that determine toxicity of endosulfan in Japanese quail and other avian species. 

Enrichment will consist of group housing for hatchlings and juveniles and pair housing for 

adults. Singly housed birds will be provided with sisal rope. 

V.3.5.2. Reduction: This study will utilize a group of birds to determine reproductive, 

developmental, immune, and behavioral effects in the same individuals. This is a large reduction 

in the number of birds relative to the number that would be needed to measure these effects 

separately in individual birds. 

V.3.5.3. Replacement: No replacement alternatives are appropriate for this study. 

V.4. Technical Methods: 

V.4.1. Pain/Distress Assessment: Pain or distress are not expected to be experienced by 

any of the birds from the treatments in this study. 

V.4.1.1. APHIS Form 7023 Information 
V.4.1.1.1. Number of animals: 440 (maximum potential ~800)

V.4.1.1.1.1. Column C: 440 – ~800 (100%) 

V.4.1.1.1.2. Column D: 0 

V.4.1.1.1.3. Column E: 0 

V.4.1.2. Pain Relief/Prevention: No other substances that could potentially interfere with the 

interpretation of these results will be administered in the unlikely event that the animal appears 

to be experiencing pain or discomfort. However, moribund or animals in overt pain will be 

humanely euthanized as described in section V.4.6. (Euthanasia). Based on previous experience 

with this model, we will consider the following as indications of distress where recovery is 

unlikely at continued exposure regimes: 

1) Loss of body weight to exceed 20%. 

2) Loss of righting response and inability to care and feed without assistance. 

3) Indications of respiratory infection (nasal discharge coupled with congested breathing). 

A weight of evidence determination will be made on a case-by-case basis after consultation with 

the principal investigator and the attending veterinarian. 

V.4.1.2.1. Anesthesia/Analgesia Tranquilization: n/a 

V.4.1.2.2. Pre- and Post-procedural Provisions: None

V.4.1.2.3. Paralytics: None

V.4.1.3. Literature Search for Alternatives to Painful or Distressful Procedures: 

V.4.1.3.1. Sources Searched:

 

 a. MEDLINE®: National Library of Medicine Database (1950 – 2008) 

b. RTECS: Registry of Toxic Effects of Chemical Substances (1960 – 2007) 

Science (1996-1999) 

c. TOXLINE® (1900 – 2006) 

d. PubMed (1980-2008) 

e. Adverse Reaction Database (2007-2008) 

f. Toxfile (1965-2008) 

g. Inside Conferences (1993-2008) 

h. Biological and Chemical Index (1983-2008) 

i. DTIC, STINET: Defense Technical Information Center Private Scientific and Technical 

Information Network (1980-2008) 

j. AGRICOLA: USDA National Agricultural Library (1970 – 2008) 

k. International Phamacology Abstracts (1970-2007) 

l. BIOSIS Toxline ® (1969-2008) 

m. Pharm-line ® (1978-2002) 
n. Pascal (1973-2008) 

o. Global Health (1983-2008) 

p. CBRN START: Chemical, Biological, Radiological and Nuclear. Scientific and 

Technical 

q. Analysis Research Tool (1980-2008) 

r. CAB Abstracts (1972-2008) 

s. EMBASE (1974-2008) 

t. NTIS: National Technical Information Service (1964-2008) 

u. BIOSIS Previews (1926-2008) 

V.4.1.3.2. Date of Search: 11 February 2008 

V.4.1.3.3. Period of Search: The period of search of the databases is the same as that stated in 

section II.2.3. 

V.4.1.3.4. Key Words of Search: endosulfan, thionex, thiodan, benzoepin, rasayansulfan, 

chlorthiepin, devisulfan, Japanese quail, quail, feed aversion, stress, preven*, control, manage*, 

reduce, elimina*, alleviat*, decreas*, diminish, relief, animal testing reduction, animal testing 

refinement, biolog* Model*, theoretical model, biolog* model*, animal testing replacement, 

animal testing alternative, alternat*, computer model*, simulate, alternative animal test*, animal 

model*. Asterisks indicate that the word roots were used as search terms. 

V.4.1.3.5. Results of Search: Few papers were found that discussed avian euthanasia 

methods. Several papers were found that evaluated various anesthesia techniques, including the 

use of halothane, ketamine-xylazine, and isoflurane. Nothing was found regarding pain or 

distress. 

Additionally, an independent literature search was conducted using a combination of several 

academic search engines and by manually tracking down literature cited in captive care 

literature. This search yielded several class-specific anesthesia/euthanasia review papers. The 

following synopses summarize our most relevant findings: 

1. There are no alternatives to using live birds for addressing the purpose of this study. 

2. The housing and husbandry design is appropriate to the study and in meeting the life 

maintenance and enrichment needs of the Japanese quail. 

3. The use of asphyxiation through CO2 exposure has been recommended for anesthesia and 

euthanasia in birds (AVMA, 2007). 

V.4.1.4. Unalleviated Painful/Distressful Procedure Justification: Since the goal of this 

investigation is to determine the effects from exposure to an environmental contaminant, no 

other substances that could potentially interfere with the interpretation of these results will be 

administered. However, moribund or animals in overt pain will be humanely euthanized as 

described in section V.4.6.
V.4.2. Prolonged Restraint: Not applicable

V.4.3. Surgery: Not applicable 

V.4.3.1. Pre-surgical Provisions: Not applicable

V.4.3.2. Procedure: Not applicable

V.4.3.3. Post-surgical Provisions: Not applicable

V.4.3.4. Location: Not applicable

V.4.3.5. Surgeon: Not applicable

V.4.3.6. Multiple Major Survival Operative Procedures: Not applicable

V.4.3.6.1. Procedures: Not applicable

V.4.3.6.2. Scientific Justification: Not applicable

V.4.4. Animal Manipulations: Quail will be restrained briefly while the injections for the 

immune response measures are administered. One person will present the bird to the injector, 

keeping the wings and legs firmly pressed against its body. The injector will then brush the 

feathers away from the featherless tract in the neck region and then inject the solution into the 

jugular vein. Blood will also be collected from this vein. 

V.4.4.1. Injections: Not applicable 

V.4.4.2. Biosamples: Blood will be collected according to DTOX SOP 053. 

V.4.4.3. Adjuvants: Chukar partridge erythrocytes.

V.4.4.4. Monoclonal Antibody (MAbs) Production: Not applicable.

V.4.4.5. Animal Identification: Individual animals will be identified by cage card and leg 

bands according to DTOX SOP 003.

V.4.4.6. Behavioral Studies: Not applicable.

V.4.4.7. Other Procedures: None. 

V.4.4.8. Tissue Sharing: Tissues may be shared with U.S. Army Corps of Engineers, 

Engineering Research and Development Center for toxicogenomics research.

V.4.5. Study Endpoint: The study will be terminated within one week via euthanasia 

following completion of reproductive behaviors assessed in the F2 generation. Early euthanasia 
may occur when a weight of evidence determination (as described in V.4.1.2) indicates that 

animals are experiencing significant pain or distress. 

V.4.6. Euthanasia: Birds will be euthanized by CO2 asphyxiation as recommended by the 

AVMA Guidelines on Euthanasia (2007) and as specified by SOP 066. Although unhatched 

birds are not covered by these guidelines, fertilized eggs that are not destined to be set to hatch 

will be frozen prior to being discarded, and late stage embryos will be decapitated immediately 

following removal from the egg. 

V.5. Veterinary Care: 

V.5.1. Husbandry Considerations: Treatments will be randomly assigned to birds within 

cages and cage units. Cage unit frames are constructed of 1” stainless steel tubing. Cages are 

constructed of perforated stainless sheet. Each cage unit will have two litter pans per tier that are 

easily removed. Each unit will have individual stainless steel feeders and one stainless Edstrom 

cup drinker. Watering will be automatic. Cage dimension is 7” wide x 9.5” tall x 11” deep. 

Birds will be acclimated to laboratory conditions for a minimum of five days prior to the onset of 

experiments. Birds will be kept in conditions of constant photoperiod. Birds will be housed in 

AAALAC accredited facility at USACHPPM. Each cage will contain a card describing 

treatment and animal number. Detailed husbandry practices are contained in DTOX SOP 004. 

The photoperiod for chicks and adults will be 16 or 17 hours light and 8 or 7 hours of darkness. 

Chicks and juveniles will be group housed, and adults of the P and F1 generations will be pair 

housed. Adult males from the F2 generation may be single housed for reproductive behavioral 

measures, and the F2 females may be single housed for the assessment of onset of reproductive 

maturity. Individuals will be paired according to similar weights to minimize the potential for 

aggression. Pairs will be monitored daily for pen mate aggression, and incompatible pairs will 

be separated and repaired as appropriate.

Hatchlings will be dried completely before being transferred to brooders. Once grouped, chicks 

will be checked at least twice daily to ensure that group shunning, hypothermia, and/or starvation 

do not occur. Sufficient space for feeding and drinking will be provided, especially during the 

first week post-hatch to allow equal access to food and water to weaker chicks. Chicks will be 

housed together by treatment group and will be identifiable individually by leg bands. Sexes will 

be separated at 4-5 weeks and placed into individual cages to allow for determination of 

reproductive maturity. Birds from the F1 generation will be pair housed following reproductive 

behavior measures. 

Housing conditions: 

Age (weeks) Temp. (o

C) RH (%) 

1 35-38 40-80 

2 30-35 40-80 

3-4 23-27 40-80 

>4 16-27 40-80 
Feed and water will be provided ad libitum. Chicks will receive a growth diet and adults will be 

given a laying diet that will accommodate for expenditure of calcium through egg laying. 

Eggs will be stored in a cold storage facility for a maximum of one week prior to setting in an 

incubator. Cool storage prevents embryo development and aids in synchronizing the 

development of embryos of eggs laid during that week. All cracked and abnormal eggs will be 

removed prior to incubation. Eggs will be equilibrated to room temperature prior to being set in 

the incubator. All cracked and abnormal eggs will be recorded. Eggs should be turned in the 

incubator at least three times a day. For incubation, temperature should be between 37.5-37.8o

C 

and relative humidity should be between 50-70%. Eggs should be hatched at 37-37.5o

C and 

relative humidity should be between 70-75%. 

V.5.1.1. Study Room: Housing- as assigned, E2100/ E-2101. 

 Dosing- rm. E2100 / E2101 

 Necropsy- rm. 3201, E2100 

V.5.1.2. Special Husbandry Provisions: Not applicable 

V.5.1.3. Exceptions: None

V.5.2. Veterinary Medical Care: 

 

V.5.2.1. Routine Veterinary Medical Care: All animals will be observed at least twice daily 

by the animal care staff and once on weekend days and holidays. Appropriate methods of animal 

care shall be maintained to prevent, control, diagnose, and treat diseases and injuries. If it is 

determined that an animal is under distress, comatose, or moribund then the observer will report 

to the principal investigator or the attending veterinarian for an examination of its disposition and 

medical condition. If the animal is judged (by the PI through consultation with the attending 

veterinarian) unlikely to recover, as indicated by having met criteria spelled out in V.4.1.2., then 

the animal will be humanely euthanized as described. 

V.5.2.2. Emergency Veterinary Medical Care: Animals will be observed at least once daily 

by the animal care staff on weekend days and holidays. If an animal is noted to be ill, the 

Attending Veterinarian will be contacted. Should the animal require euthanasia as described 

previously, the Attending Veterinarian will contact the PI/study director. If the PI/study director 

or other co-investigators are not available, the Attending Veterinarian will euthanize the animal 

and take as many required samples as possible from the animal according to protocol. The 

animal carcass will be held at the facility under refrigeration for a minimum of three days in 

order for investigators to take any additional required samples.

V.5.3. Environmental Enrichment: 

V.5.3.1. Enrichment Strategy: Enrichment will consist of group housing for hatchlings and 

juveniles and pair housing for adults. Singly housed birds will be provided with sisal rope.

V.5.3.2. Enrichment Restriction: NONE. 
VI. STUDY PERSONNEL QUALIFICATIONS AND TRAINING: 

 All investigators named in this study have demonstrated an understanding of the humane care 

and use of research animals. They have taken part in discussions of pertinent laws and 

regulations concerning the use of animals in biomedical research in the Department of Defense 

as required by Public Laws 89-544, 91-579, and 99-198 (The Animal Welfare Act and 

Amendments, DOD Directives, and Army Regulations). They are familiar with the concepts for 

the reduction or elimination of the use of animals and have concluded that there is a need for the 

use of animals in this study. They have been familiarized in the proper methods for minimizing 

and/or alleviating pain in the animal species selected for study. 

 If their training in required procedures has not been of a formal, documented nature, they will 

demonstrate proficiency in these procedures prior to performing them on study animals. The 

Attending Veterinarian will observe them and determine proficiency or additional training needs. 

 Personnel working on this study have been advised on the animal care and use policy at this 

institution and are aware of the established reporting mechanisms for observed deficiencies in 

animal care and treatment. The following is a table of participating investigators listing training 

and qualifications: 

Study Personnel Table 

Staff Member Procedure(s) Years of Experience Qualifications 

Quinn Collection and 

handling, 

phlebotomy, 

euthanasia, 

necropsy 

10+ yrs animal research. 

PI and Co-PI on studies using 

the northern bobwhite and 

Japanese quail; Quail 

necropsy training (June 2005) 

Ph.D, Animal 

Science 

Johnson Handling, 

euthanasia, 

necropsy, dosing 

10+ yrs animal research. 

Refined the bobwhite model 

for its use in ecotoxicology 

assessment; Short course on 

animal care & use (May 

2000, April 2003) 

Ph.D, Veterinary 

Medical Science; 

D.A.B.T., 

McFarland Handling, 

euthanasia, 

necropsy, dosing 

10+ years animal and 

toxicological investigations: 

Northern bobwhite handling, 

dosing, prosection training 

(Sept 2005); Avian handling 

& blood withdrawal (Jan 

2007) 

Ph.D, Biology, 

DVM 

Amanda HessRuth Handling, 

behavior, 

necropsy, dosing 

5+ years animal and 

toxicological investigations, 

experience specific to animal 

behavior; will receive 

specific OTJ training 

B.S. Animal 

Behavior 

LaFiandra Handling, 

euthanasia, 

10 years animal and 

toxicological investigations; 

Ph.D, Natural 

Resources and 
necropsy, dosing Avian euthanasia & necropsy 

training (Dec 2007); Avian 

Handling & Dosing (in-house 

training, Dec 2007- Mar 

2008) 

Environmental 

Studies 

Dr. Michael Quinn has11 years of experience in avian toxicology and has over 20 publications in 

the field of wildlife toxicology. He has worked in this field at the Patuxent Wildlife Research 

Center USGS, Shippensburg University, and University of Maryland, and has worked at CHPPM 

for the last three years. Dr. Mark Johnson has a background in population ecology and 

veterinary medical science. He has worked in the area of environmental risk assessment and has 

over 30 publications in the field of wildlife toxicology and ecology. He has been working in this 

field at CHPPM for the last 15 years. Dr. Craig McFarland has graduate training and experience 

in wildlife biology (MS) and environmental toxicology (PhD). He has 10 years field and 

laboratory experience in studies of small mammal toxicology and was a practicing small animal 

veterinarian (DVM) for 20 years. Amanda Hess-Ruth has over five years of experience in 

animal research specific to toxicology and has demonstrated expertise in assessing animal 

behavior. Dr. Emily LaFiandra has been a toxicologist here at CHPPM for the past year and has 

gained extensive on the job experience handling bobwhite. 

 

VII. BIOHAZARD/SAFETY:

Fowl of this type have been reported to carry specific zoonotic diseases that can be 

transferred to humans (e.g., psittacosis, histoplamosis), most of them affecting the respiratory 

system. The flock from which the eggs will come has been maintained and screened for diseases 

for over 15 years. Adherence to standard chemical and animal handling procedures will be 

required during the performance of this study as indicated in CHPPM Regulations 385-1 and 

385-5 and DTOX SOP 083. All personnel will use paper respirators, latex or nitrile gloves, and 

lab coats when handling the birds or endosulfan-treated feed. 

VIII. ENCLOSURES: References 

 CHPPM Standard Operating Procedures 

IX. STUDY TIME FRAME: 

IX.1. Estimated Experimental Initiation Date: April 2008 

IX.2. Estimated Experimental Completion Date: February 2009 
X. ASSURANCES: 

As the Study Director/Principal Investigator on this protocol, I acknowledge my 

responsibilities and provide assurances for the following: 

A. Animal Use: The animals authorized for use in this protocol will be used only in the 

activities and in the manner described herein, unless a modification is specifically approved by 

the IACUC prior to its implementation.

B. Duplication of Effort: I have made every effort to ensure that this protocol is not an 

unnecessary duplication of previous experiments. 

C. Statistical Assurance: : I assure that I have consulted with a qualified individual 

who evaluated the experimental design with respect to the statistical analysis, and that the 

minimum number of animals needed for scientific validity will be used. 

D. Biohazard/Safety: I have taken into consideration and made the proper 

coordinations regarding all applicable rules and regulations concerning radiation protection, 

biosafety, recombinant issues, and so forth, in the preparation of this protocol. 

E. Training: I verify that the personnel performing the animal 

procedures/manipulations/observations described in this protocol are technically competent and 

have been properly trained to ensure that no unnecessary pain or distress will be caused to the 

animals as a result of the procedures/manipulations. 

F. Responsibility: I acknowledge the inherent moral, ethical and administrative 

obligations associated with the performance of this animal use protocol, and I assure that all 

individuals associated with this project will demonstrate a concern for the health, comfort, 

welfare, and well-being of the research animals. Additionally, I pledge to conduct this study in 

the spirit of the fourth "R," namely "Responsibility," which the DOD has embraced for 

implementing animal use alternatives where feasible and conducting humane and lawful 

research. 

G. Scientific Review: This proposed animal use protocol has received appropriate peer 

scientific review and is consistent with good scientific research practice. 

H. Painful Procedures: I am conducting biomedical experiments, which may 

potentially cause more than momentary or slight pain or distress to animals. This potential pain 

and/or distress WILL NOT be relieved with the use of anesthetics, analgesics, and/or 

tranquilizers. I have considered alternatives to such procedures; however, I have determined that 

alternative procedures are not available to accomplish the objectives of this proposed 

experiment. 

 

Michael J. Quinn, Jr. 

 _________________________________ 

Signature Date (YYYYMMMDD)
Animal Use Protocol-Two-generation test for endocrine disruption & reproductive toxicity in the Japanese quail with 

endosulfan. 

 

APPENDIX A 

References:

Dunachie, J.F. and W.W. Fletcher. 1966. Effect of some insecticides on the hatching rate of 

hens’ eggs. Nature, 212: 1062. 

Dunachie and W.W. Fletcher. 1969. An investigation of the toxicity of insecticides to birds’ 

eggs using the egg injection technique. Annu. Appl. Biol., 64: 409-423. 

EPA (U.S. Environmental Protection Agency). 1996. Avian reproduction test. OPPTS 

850.2300 “Public Draft” Ecological Effects Test Guidelines. EPA 712-C-96-141. U.S. 

Environmental Protection Agency, Washington, D.C. 

FWS (U.S. Fish and Wildlife Service). 1975. Lethal Dietary Toxicities of Environmental 

Pollutants to Birds. ., U.S. Fish and Wildlife Service, Bureau of Sports Fisheries and 

Wildlife. Special Scientific Report - Wildlife No. 191. Washington, DC, U.S. 

Garg, U.K., A.K. Pal, G.J. Jha, S.B. Jadhao. 2004a. Haemato-biochemical and immunopathophysiological effects of chronic toxicity with synthetic pyrethroid, organophosphate 

and chlorinated pesticides in broiler chicks. Int. Immunopharmacol., 4: 1709-22. 

Garg, U.K., A.K. Pal, G.J. Jha, S.B. Jadhao. 2004b. Pathophysiological effects of chronic 

toxicity with synthetic pyrethroid, organophosphate and chlorinated pesticides on bone 

health of broiler chicks. Toxicol Pathol., 32: 364-9. 

Hill, E. F. and Camardese, M. B. 1986. Lethal Dietary Toxicities of Environmental 

Contaminants to Coturnix, Technical Report Number 2. U.S. Department of Interior, Fish 

and Wildlife Service, Washington, DC, 6-55. 

Hudson, R.H., R.K. Tucker, M.A. Haegele. 1972. Effect of age on sensitivity: acute oral 

toxicity of 14 pesticides to mallard ducks of several ages. Toxicol. Appl. Pharmacol., 22: 

556-561. 

Hudson, R.H., R.K.Tucker, M.A. Haegele. 1984. Handbook of Acute Toxicity of Pesticides 

to Wildlife, Resource Publication 153. U.S. Department of Interior, Fish and Wildlife 

Service, Washington, DC, 6-54. 

Lutz, H. and Y. Lutz-Ostertag. 1972. The action of different pesticides on the development of 

bird embryos. Adv. Exp. Med. Biol., 27: 127-150. 

Matthews, J., T. Celius, R. Halgren, T. Zacharewski. 2000. Differential estrogen receptor 

binding of estrogenic substances: a species comparison. J. Steroid Biochem. Mol. Biol., 

74: 223-234. 
Animal Use Protocol-Two-generation test for endocrine disruption & reproductive toxicity in the Japanese quail with endosulfan.

 

APPENDIX B 

CHPPM Standard Operating Procedures and Regulations 

SOP No. 003.07, Individual Animal Identification. 

SOP No. 004.07, Animal Facilities and Caretaker Duties. 

SOP No. 053.07, Animal Bleeding Technique. 

SOP No. 066.07, Animal Euthanasia. 

SOP No. 083.07, Health and Safety of Laboratory Personnel. 

USACHPPM Regulation 385-1, Safety and Occupational Health Program, May 01, 2001. 

USACHPPM Regulation 385-5, Occupational Health and Safety of Animal Users, June 1, 2007. 
22 

Protocol Modifications: 
23 
24 
25 
26 
27 
28 
29 
30 
31 
32 
33 
34 
35 

APPENDIX E 

DEVIATIONS TO THE PROTOCOL 

1. Efforts to include measures of immune function were withdrawn from the study 

design due to a lack of available chukar partridge red blood cells needed to 

assess humoral response. The customer decided that measures specific to the 

endocrine and reproductive systems be given priority over those of the immune 

system. 

2. Despite numerous efforts, sample volumes of blood from day old chicks and late 

stage embryos required for hormone and vitellogenin analyses could not be 

collected. 

3. Although our chemists were able to measure endosulfan in egg yolk, they were 

unable to detect hormones in this matrix. 

4. Vitellogenin in adult blood could not be measured. Efforts to develop methods to 

measure vitellogenin via PCR were developed but not able to be included in this 

study because since it was not anticipated, tissues were not flash frozen upon 

collection. 

5. The protocol states that eggs should be candled for fertility after 8 days of 

incubation and again for viability of embryos approximately 2 days before 

hatching. Fertility and viability were determined by candling and/or removal of 

contents after all chicks were hatched. In this way, determination of embryo age 

(week 1 and 2) was easy and prevented disturbance of embryos during 

development. 

6. The protocol stated that “during the treatment periods, one egg per pen will be 

collected from odd numbered pens in odd numbered weeks and from even 

numbered pens during even numbered weeks. These eggs will be used to 

assess eggshell strength and eggshell thickness.” We instead measured 

eggshell strength and thickness from two eggs per hen (when available) that 

were produced within the same week. 

7. Epididymides were collected and used for sperm analysis. As stated in the 

protocol, “Histopathology will be performed on the brain, right thyroid gland, liver, 

bursa of Fabricius, kidneys, and gonads.” Epididymides were not, therefore, 

included in the histopathological analysis. 

8. Gender determination could not be completed for all young chicks found dead 

while in the brooders, and since there is no evidence of EDCs changing gender 

in birds it was agreed that the sex ratios of F1 and F2 chicks would not be 

included in this study. 

9. Histopathology was performed by Experimental Pathology Laboratories, Inc., 

Sterling, VA. Their results have been sent to the EPA under a separate Scope of 

Work and will not be included in this report. 
36 

APPENDIX F 

DIET PREPARATION REPORT FROM 

WILDLIFE INTERNATIONAL, LTD. 
37 

Wildlife International, Ltd. Project Number 167-142 

Test Substance 

 The test substance, endosulfan, was received from Helena Industries, Cordela, GA 31015 

on December 12, 2008 and was assigned Wildlife International, Ltd. identification number 8838 

upon receipt. The test substance was a solid and was identified in the accompanying 

correspondence as: endosulfan 95%. The test material had a reported purity of 95%. The test 

substance was held under ambient conditions in locked storage at the Wildlife International, Ltd. 

facilities in Easton, Maryland. Concentrations of the test substance in the diet were “as is” and 

not adjusted to 100% active ingredient. Therefore, dietary concentrations are expressed as parts 

per million (ppm) in the diet. 

Diet Preparation 

 The basal diet fed to both adults and offspring was formulated to Wildlife International, 

Ltd. specifications by Cargill Animal Nutrition, Shippensburg, PA (Appendix I). The basal 

ration contained at least 27% protein and 2.5% crude fat, and no more than 3.8% crude fiber. 

The basal diet contained approximately 1.0% calcium, derived from feedstuffs and the 1.0% 

limestone used in the formulation of the basal diet by Cargill. While this level of calcium is 

sufficient for growth and maintenance rations, additional calcium is required in the ration of 

breeding birds for egg shell formation. Therefore, an additional 5% (w/w) of limestone 

(approximately 38.5% Ca) was added to the basal diet for the adults. This raised the calcium 

level in the diet for the breeding birds to approximately 3%, slightly above the minimum 

recommended for quail. Offspring received diets prepared without the addition of 5% 

supplemental limestone. 

 Test diets were prepared by mixing endosulfan into a premix that was used for preparation 

of the final diet. Control diet and each of the four treated diets were prepared on December 23, 

2008; March 28, 2009, June 22 & 23, 2009 and August 15, 2009. Dietary concentrations were 
38 

not adjusted for purity of the test substance and are presented as parts per million (ppm). Details 

of the preparation of test and control diets are shown in Appendix II. 
39 

Appendix I 

Diet Formulation 

WILDLIFE INTERNATIONAL, LTD. GAME BIRD RATION1 

INGREDIENTS

Fine Corn Meal 

Soy Bean Meal, 47.5% Protein 

Wheat Midds 

Agway Special, 60% Protein 

Alfalfa Meal, 20% Protein 

Dried Whey 

Ground Limestone 

Eastman CalPhos 

GL Ferm (Fermatco)2

Salt Iodized 

CM T-Premix 561 

CM TM Premix 434 

CHO CHL 70% 

Liquimeth 40% 

Lysine 75% 

DL Methionine 055 

Vit K 16 g/lb 50# 

Selenium 

Total 

PERCENT (%)

42.86 

33.39 

6.48 

9.96 

3.00 

2.50 

0.62 

0.11 

0.25 

0.10 

0.05 

0.08 

0.09 

0.25 

0.06 

0.10 

0.05 

0.05 

100.00 

VITAMIN AND MINERAL CONTENT AMOUNT ADDED PER 

POUND 

Vitamin D3 Vitamin A 

Riboflavin 

Niacin 

Pantothenic Acid 

Vitamin B12 Folic Acid 

Biotin 

Pyridoxine 

Thiamine 

Vitamin E 

Vitamin K (Menadione Dimethylpyrimidinol 

Bisulfite) 

Manganese 

Zinc 

Copper 

Iodine 

Iron 

Selenium 

Beta-Carotene 

Calcium 

Chloride 

Choline 

Cobalt 

Magnesium 

Phos 30 

Phos 50 

Potassium 

Sodium 

Sulfur 

Phosphorus 

Ca:PHOS 

0.600 kiu/lb

 2.000 kiu/lb 

 2.1992 mg/lb 

 21.6932 mg/lb 

 5.2241 mg/lb 

 6.0000 ug/lb 

 0.5169 mg/lb 

 65.9360 ug/lb 

 1.4639 mg/lb 

 0.9200 mg/lb 

 5.0000 iu/lb 

 3.2400 mg/lb 

 115.0203 mg/kg 

 194.6601 mg/kg 

 24.3007 mg/kg 

 2.4410 mg/kg 

 150.4117 mg/kg 

 0.4749 mg/kg 

 1.1413 mg/lb 

 0.9750 % 

 0.2164 % 

 1882.4116 mg/kg 

 0.1139 mg/kg 

 0.2503 % 

 0.3908 % 

 0.4719 % 

 1.1148 % 

 0.1335 % 

 0.3478 % 

 0.6800 % 

 1.4338 ratio 

1

 The guaranteed analysis is a minimum of 27% protein, a minimum of 2.5% crude fat and a 

maximum of 3.8% crude fiber. 2

 Fermentation By-Products (Source of Unidentified Growth Factors) 

 
40 

Appendix II 

Diet Preparation 

 Premixes for endosulfan were prepared on December 22, 2008; March 27, 2009; June 21, 

2009 and August 11, 2009. Nominal preparation was as follows: 

 Control: 4018.0 g ration + 170 ml acetone + 91 ml corn oil 

 30 ppm: 3.3825 g Endosulfan + 4014.6 g ration + 170 ml acetone + 91 ml corn oil 

 60 ppm: 6.7650 g Endosulfan + 4011.2 g ration + 170 ml acetone + 91 ml corn oil 

 125 ppm: 14.0938 g Endosulfan + 4003.9 g ration + 170 ml acetone + 91 ml corn oil 

 250 ppm: 28.1875 g Endosulfan + 3989.8 g ration + 170 ml acetone + 91 ml corn oil 

For each premix, the appropriate amount of endosulfan was weighed in a tared plastic weigh 

boat on an analytical balance. The test substance was transferred to a 400 ml beaker. Acetone 

was measured in a graduated syringe and added to the test substance with a portion of the 

acetone used to rinse the weigh boat. The rinse was added to the beaker. A magnetic stir-bar 

was placed in the beaker and the mixture stirred for at least two minutes (approximate), on a 

magnetic stir plate, until the endosulfan dissolved. Corn oil was then measured in a graduated 

cylinder and added to the test substance solution. Mixing then was continued for at least one 

additional minute. 

The required basal ration was weighed into a tared mixing bowl on a top-loading balance. 

The test substance mixture was then added to the basal ration in the mixing bowl. The beaker 

was rinsed with additional acetone measured in a graduated syringe. The rinse was added to the 

mixing bowl and the contents of the bowl mixed on a stand mixer for approximately 20 minutes. 

After mixing, 2000.0 g aliquots of the premix were weighed on a top-loading balance. Unless 

used immediately for preparation of the final diets, the premixes were placed in appropriately 

labeled plastic bags, reweighed and stored in a freezer. 

 

 As needed, the appropriate premix was incorporated into the final diet for adult birds as 

follows: 

 0 ppm: 2000 g Premix + 50.250 kg ration + 2750 g limestone 

 30 ppm: 2000 g Premix + 50.250 kg ration + 2750 g limestone 

 60 ppm: 2000 g Premix + 50.250 kg ration + 2750 g limestone 

 125 ppm: 2000 g Premix + 50.250 kg ration + 2750 g limestone 

 250 ppm: 2000 g Premix + 50.250 kg ration + 2750 g limestone 

 

 As needed, the appropriate premix was incorporated into the final diet for chicks as 

follows: 

 0 ppm: 2000 g Premix + 53.000 kg ration 

 30 ppm: 2000 g Premix + 53.000 kg ration 

 60 ppm: 2000 g Premix + 53.000 kg ration 

 125 ppm: 2000 g Premix + 53.000 kg ration 

 250 ppm: 2000 g Premix + 53.000 kg ration 
41 

 The diet was mixed for approximately twenty minutes in a Patterson-Kelly® Twin Shell 

Blender. 

APPENDIX G 
42 

CONTRIBUTING SCIENTIST REPORT 

STATISTICAL ANALYSIS OF DATA 
43 

Statistical Analysis on USAPHC Toxicology Protocol 

Number: 08JG-80-08-04-02 

Two-Generation Test for Endocrine Disruption and Reproductive Toxicity in 

Japanese Quail (Corturnix japonica) using Endosulfan 

Good Laboratory Practice Compliance Statement 

The statistical analysis of the data for the Two-generation test for endocrine disruption 

and reproductive toxicity in Japanese quail (Cortunix japonica) using endosulfan was 

conducted in compliance with Title 40, Code of Federal Regulations (CFR), Part 792, 

Good Laboratory Practice Standards. Statistical analyses were conducted according to 

the procedures outlined in the protocol for this study (Protocol Number 08JG-80-08-04-

02). 

 

 17 February, 2012 

Shane Hall Date 

Statistician 

 
44 

Table of Contents:

ENDOSULFAN ENDOCRINE DISRUPTION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

. . . . . . . . . . . . 3 

Eggshell Thickness and Strength . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

. . . . . .4 

Egg Production . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

. . . . . 6 

Sperm . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

. . . . . .9 

Growth and Body Weight . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

. . . . 11 

Feed Consumption . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

. . . . 15 

Cloaca Gland (CG) Area . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

. . . . . 19 

Yolk Content . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

. . . . .21 

Onset to Puberty . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

. . . . 22 

ENDOSULFAN EFFECTS ON REPRODUCTIVE BEHAVIOR . . . . . . . . . . . . . . . . . . . . . . . 

. . . . . . . . . . . . . . 24 

Number of Mount Attempts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

. . . 25 

Time Until First Mount . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

. . . . 27 

Number of Copulations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

. . . 29 

Time Until First Copulation . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

. . . . 31 

ENDOSULFAN EFFECTS ON ORGAN WEIGHTS . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . 

. . . . . . . . . . . . 33 

 
45 

ENDOSULFAN ENDOCRINE DISRUPTION

Egg, Sperm, and Growth Characteristics 

This report analyzes the effects that endosulfan had on the endocrine system of male and 

female quail through three generations (F0, F1, and F2). Data was collected on egg strength and 

thickness, egg production, sperm characteristics, body weights, food consumption, coacal gland 

(CG) area, P generation yolk content, and onset to puberty. All quails were part of one of five 

dose groups: (control) 0 ppm, 30 ppm, 60 ppm, 125 ppm, and 250 ppm. For endocrine 

disruption measures that contained weekly measurements, the same quails were used from week 

to week. SPSS version 16.0 was used to analyze the behavior data. Statistical significance for 

this analysis was determined using an alpha (α) of .05 with a 95% confidence level. 

 
46 

EGGSHELL THICKNESS AND STRENGTH 

I. Analysis Methods 

 An Analysis of Variance (ANOVA) test was used to determine if the average eggshell 

thickness and/or average eggshell strength for the dose groups were significantly different. An 

F-statistic and corresponding p-value were calculated for both thickness and strength. If the pvalue was below .05, then it was concluded that at least two dose groups had statistically 

different averages for that measure. A Tukey post-hoc test was then used to find which specific 

dose groups were statistically different from one another. A Levene’s test was used to check to 

make sure the dose groups had equal variances, a major assumption that must be met in order to 

properly run an ANOVA model. If this assumption was violated (p-value for Levene’s test < 

.05), a non-parametric Kruskal-Wallis test was run instead of an ANOVA. 

II. Summary of Results 

The general trend for the egg strength data was that as thickness increased, strength 

decreased. Larger thickness values tended to be associated with the control group and less 

concentrated endosulfan dose groups. The F1 generation had more variation between dose 

groups compared to the P and F2 generations, especially on thickness. The control group had the 

lowest average strength for all three generations, and the highest average thickness for the P and 

F1 generations. Strength averages were consistent across the three generations but thickness 

averages slightly decreased from the F0 to F1 generation, and greatly decreased from the F1 to 

F2 generation. 

III. F0 Generation Results 

No statistically significant differences occurred for thickness averages, averages were 

highest for the control group and dose 60. Dose 125 had the lowest average thickness; however 

it had the highest strength average. The control group’s average strength was directionally lower 

than all other dose groups, and was statistically significantly lower than dose 125. Strength 

averages generally increased as dose concentration increased. 

IV. F1 Generation Results 

 Statistically significant differences between doses occurred for average thickness and average 

strength. The control group’s average thickness was directionally greater than all other dose 

groups, and was significantly greater than dose 125 and dose 250. Dose 30’s thickness was also 

Dose Average Std. Error Average Std. Error

0 0.932 0.074 0.93 0.07

30 0.890 0.060 1.05 0.06

60 0.981 0.146 1.05 0.05

125 0.846 0.051 1.19 0.05

250 0.869 0.066 1.11 0.06

F0Generation ‐ Egg Strength

p‐value = .788

* Tukey Pos t‐hoc resul ts i ndica te tha t the control group

had signi fi cantly lower s trength compa red to dose group

125.

p‐value = .045*

Thickness Strength
47 

significantly greater than dose group 250. As for strength, dose 250 had a significantly higher 

average compared to the control group. Just like the F0 generation, strength averages increased 

and thickness averages decreased as endosulfan concentration increased. 

V. F2 Generation Results 

Minimal, non-significant differences (p-values >.05) occurred for thickness and strength 

averages between dose groups. Negligible fluctuation occurred in thickness data across dose 

groups, but dose 60 had the highest average thickness. Dose 30, followed by dose 250 had the 

highest average strengths. Unlike the other two generations where strength increased and 

thickness decreased as dose concentration increased, no true pattern existed between dose group 

concentration and strength or thickness for the F2 generation. 

Dose Average Std. Error Average Std. Error

0 0.938 0.052 0.96 0.05

30 0.845 0.047 1.02 0.05

60 0.813 0.052 1.04 0.04

125 0.753 0.032 1.09 0.05

250 0.647 0.038 1.19 0.04

p‐value = .000* p‐value = .016**

F1 Generation ‐ Egg Strength

Thickness Strength

* Tukey Pos t‐hoc res ul ts i ndica te tha t the control group

had signi fica ntly higher thicknes s compa red to dose 250

and dose 125.  Dose 30 also grea ter thicknes s compa red to

dose 250.

** Tukey Pos t‐hoc res ul ts i ndica te tha t the control group

had signi fica ntly lower s trength compa red to dose 250.

Dose Average Std. Error Average Std. Error

0 0.476 0.024 0.89 0.06

30 0.486 0.024 1.28 0.17

60 0.500 0.031 1.03 0.07

125 0.471 0.020 1.00 0.05

250 0.483 0.021 1.14 0.06

F2 Generation ‐ Egg Strength

Thickness Strength

p‐value = .924 p‐value = .053
EGG PRODUCTION 

I. Analysis Methods 

A Generalized Estimating Equation – poisson counts model (GEE) was used to determine if there was a signif

among dose groups in their egg production, accounting for the variability from week to week. P-values were calweek as well as their interaction. Dose, week, and interaction were determined to be significant if the p-value fo

than .05. Estimated marginal means were calculated for both factors and examined for differences using a Tukey 

p-value for that factor was less than .05. If the interaction was statistically significant, the interaction was investimeans. The significance results of the primary factors are considered to be uninformative and can be ignored. 

II. Summary of Results Egg production increased for the first few weeks of all three generations. After the initial few weeks of each gen

production from week to week was very similar. Interactions were significant for all three generations, meaning 

egg production change from week to week were not statistically equal. Apparent differences in at least one of thoccurred in the F0 and F2 generations. The control group had a much lower egg production compared to the othin the F0 generation, and dose 250 had a much lower egg production compared to the other doses after week 7 in All generations generally had the same amount of egg production. III. F0 Generation Results  The dose by week interaction was statistically significant for the F0 generation because of the decline in egg profor the control group (the other dose groups had either increases or little variation from week to week after week 125 typically had the highest egg production after the first initial weeks. 

Model Term P‐value Dose Week 6 Week 7 Week 8 Week 9 Week 10 Week 11 Week 12 Week 13 Week 14 Week 15 Week 16 We

0 0.45 2.00 3.35 4.65 4.50 3.55 3.15 2.90 3.20 2.85 3.00 2

30 0.60 2.95 3.80 4.30 4.30 5.20 5.85 6.35 6.65 6.15 6.00 660 0.40 2.40 3.80 4.25 4.55 4.95 4.70 5.60 5.10 5.40 5.55 5

125 0.05 2.05 4.35 5.45 5.95 5.35 6.20 5.95 5.60 5.75 6.05 6250 0.40 2.45 4.15 5.35 5.40 4.75 5.25 5.10 5.75 5.40 4.65 5

Overall 0.29 2.35 3.87 4.77 4.90 4.71 4.90 5.00 5.11 4.94 4.90 5

GEE Estimated Marginal Means

F0Generation Egg Production

GEE Significance Tests

Interaction ‐            

Dose by week

0.000*

Dose 0.051

Week 0.000

* Statistically significant interaction (p‐value <.05) indicated that egg production change from week to week was signi ficantly di fferent between dose groups.
IV. F1 Generation ResultsThe dose by week interaction was once again statistically significant. The interaction was significant because of ordering of dose groups for egg production. Variation between the doses for each week was much less in the F1 to the F0 and F2 generations. However, no dose group consistently had the highest or lowest egg production. 

V. F2 Generation Results 

As was in the F0 and F1 generations, the dose by week interaction was statistically significant. The interaction w

because the increase in egg production for dose 250 was not as high as the increase in the other four generations. consistently had the lowest egg production after week 8. There was no dose group that consistently had the highthrough the entire F2 generation. Egg production averages for the first few weeks of the F2 generation were low

the previous two generations. 

Model Term P‐value Dose Week 6 Week 7 Week 8 Week 9 Week 10 Week 11 Week 12 Week 13 Week 14 Week 15 Week 16 Week 17 Week 18 Week 1

0 0.95 2.80 4.45 5.45 5.40 5.70 4.90 5.20 4.50 5.30 5.90 5.90 5.30 5.20 

30 0.10 1.60 3.50 5.50 6.05 6.30 5.55 5.35 5.00 6.05 5.80 5.60 5.75 5.45 60 1.55 3.50 4.95 5.65 5.40 5.70 5.30 3.95 4.50 5.50 5.55 5.40 5.00 4.30 

125 0.25 2.35 4.20 5.60 5.70 6.05 5.55 4.95 4.15 5.00 5.20 5.10 5.95 5.90 250 0.20 3.00 4.95 5.85 5.60 5.45 4.95 5.40 4.35 5.50 5.60 5.25 5.50 5.15 

Overall 0.37 2.56 4.37 5.61 5.62 5.83 5.24 4.94 4.49 5.46 5.60 5.44 5.49 5.17 

GEE Estimated Marginal Means

F1Generation Egg Production

Dose 0.539

Week 0.000

Interaction ‐            

Dose by week

0.000*

GEE Significance Tests

* Statistically significant interaction (p‐value <.05) indicated that egg production change from week to week was significantly different between dose groups.

Model Term P‐value Dose Week 6 Week 7 Week 8 Week 9 Week 10 Week 11 Week 12 Week 13 Week 14 Week 15 Week 16 Week 17 Week 18 We

0 0.35 0.85 3.25 3.95 3.85 4.70 5.95 6.20 5.45 5.70 5.35 5.35 5.85 5

30 0.60 1.60 3.30 3.80 4.35 5.15 5.80 6.35 5.30 6.25 5.70 5.80 5.65 560 0.45 0.75 3.15 3.85 4.75 5.10 5.65 6.50 5.15 6.05 5.40 5.85 5.65 5

125 0.05 0.55 2.95 3.40 4.50 5.40 4.95 6.05 4.90 5.15 4.35 5.00 5.90 5250 0.30 1.00 2.55 2.40 3.45 4.00 3.65 4.50 4.25 4.85 4.05 4.30 4.80 4

Overall 0.27 0.89 3.03 3.43 4.15 4.84 5.12 5.87 4.99 5.57 4.93 5.22 5.55 5

* Statistically signi ficant interaction (p‐value <.05) indicated that egg production change from week to week was signi ficantly di fferent between dose groups.

GEE Estimated Marginal Means

F2 Generation Egg Production

Dose 0.087

Week 0.000

GEE Significance Tests

Interaction ‐            

Dose by week

0.000*
 

SPERM 

I. Analysis Methods 

 An Analysis of Variance (ANOVA) test was used to determine if the averages 

for percent motile, percent progressive, and sperm per gram for the dose groups 

were significantly different. An F-statistic and corresponding p-value were 

calculated for percent motile, percent progressive, and sperm per gram. If the pvalue was below .05, then it was concluded that at least two dose groups had 

statistically different averages for that corresponding measure. A Tukey post-hoc 

test was then used to find which specific dose groups were statistically different 

from one another. A Levene’s test was used to check to make sure the dose 

groups had equal variances, a major assumption that must be met in order to 

properly run an ANOVA model. If this assumption was violated (p-value for 

Levene’s test < .05), a non-parametric Kruskal-Wallis test was run instead of an 

ANOVA. 

II. Summary of Results 

 The F1 generation had severely lower averages for all three measures 

compared to the F0 and F2 generations. The F0 generation had slightly higher 

averages for percent motile and percent progressive compared to the F2 

generation, however, in general, sperm per gram averages were higher for the F2 

generation. The main exception was the control group which had about 1,400 

units more in the F0 generation compared to the F2 generation. Through all three 

generations, differences in the three sperm measures comparing across all dose 

groups was minimal, with very few statistically significant differences. 

III. F0 Generation Results 

 Only sperm per gram had significant differences among dose groups. The 

control group had significantly higher sperm per gram average than dose 125. 

The control group, followed by dose 30, had the highest sperm per gram averages 

for all dose groups. Minimal, non-significant differences occurred between the 

five dose groups for percent motile or percent progressive. The control group had 

the highest percent motile and the second highest percent progressive out of all 

the dose groups. 
2 

IV. F1 Generation Results 

Statistically significant differences between dose groups occurred only for 

percent progressive in the F1 generation. Dose 250 had a significantly higher 

average percent progressive compared to dose 60, 30, and the control group. 

Progressive and motile percents, in general, were higher in the more concentrated 

endosulfan dose groups. F1 was the only generation where this trend held true. 

Sperm per gram averages were highest for dose 60 and lowest for dose 30. 

V. F2 Generation Results 

 In the F2 generation, percent progressive was the only factor showing a 

significant difference. The control group and dose group 60 had a significantly 

greater percent progressive compared to the other three dose groups. No 

significant differences occurred between dose groups for either percent motile and 

sperm per gram, but dose 60 and dose 125 had the two highest averages. 

Dose Average Average Average

0 93.5 17.6 6917.7

30 84.3 17.3 5278.7

60 89.9 16.8 4786.7

125 86.8 18.1 4212.0

250 88.4 16.7 5132.3

1.2

7.9

2.1

4.0

3.3

1.7

2.2

1.3

2.6

1.7

385.4

786.3

Std. Error Std. Error Std. Error

709.7

610.7

443.6

F0 Generation ‐ Sperm

% Motile Sperm/gram % Progressive

* Tukey Pos t‐hoc tes t indi ca tes tha t dose 125 has a signi ficantly di ffe rent s pe rm per

gram ave rage compa red to the control group.

p‐value = .600 p‐value = .983 p‐value = .033*

Dose Average Average Average

0 37.3 3.8 443.2

30 31.6 3.1 342.3

60 45.3 5.5 496.2

125 44.4 4.5 427.7

250 47.8 9.8 465.2

° ID 177 (dose 250) excl uded from a nalysi s: recorded 0's for all three mea s urements

* Tukey post‐hoc tes t i ndi ca ted tha t dose 250 has signi fi ca ntly grea te r pe rcent

progres sive average compa red to dose 60, 30, and the control group.

1.5

Std. Error

% Progressive

56.6

55.0

46.0

Std. Error Std. Error

1.0

1.3

8.2

5.6

p‐value = .328 p‐value = .010* p‐value = .338

60.4

53.0

0.8

1.9

5.9

5.9

3.5

Sperm/gram

F1 Generation ‐ Sperm

% Motile
3 

 

Dose Average Average Average

0 77.8 16.0 5585.6

30 78.4 12.3 5167.8

60 85.4 15.6 6879.8

125 81.4 12.2 5663.6

250 66.2 11.6 5284.2

Std. Error Std. Error Std. Error

4.0

3.3

1.4

1.8

6.1

698.2

856.9

579.1

374.8

653.5

1.6

1.5

0.8

0.8

1.7

p‐value = .338* p‐value = .014*° p‐value = .303*

% Motile % Progressive Sperm/gram

° Pos t hoc eval ua tion indi ca tes tha t the control group and dose 60 have signi ficantly

grea ter percent progres sive compa red to dose 30, 125, and 250.

* P‐val ue calcula ted from Kruskall‐wallis tes t ‐ homogenei ty of va riance a s s umpti on

viola ted for pa rame tri c ANOVA tes t    

F2Generation ‐ Sperm
4 

GROWTH and BODY WEIGHT 

I. Analysis Methods 

A repeated measured Linear Mixed Model was run to determine if there was a 

significant difference in chick body weights among the five dose groups. A 

repeated measures model was used to test the dose and week factors as well as 

their interaction, while accounting for the variation in the data from week to 

week. Dose, week, and interaction were determined to be significant if the pvalue for the Mixed Model test was less than .05. Estimated marginal means were 

calculated for both factors and examined for differences using a Tukey post-hoc 

test, if the p-value for that factor was less than .05. If the interaction was 

statistically significant, the interaction was investigated via marginal means. The 

significance results of the primary factors are considered to be uninformative and 

can be ignored. 

 ANOVA tests were used to determine differences between doses in single 

measurement-body weight, tarsus length, and length from the tip of the beak to 

the culmen averages. Since the same birds were not measured each week, 

ANOVA tests needed to be performed separately for each day of measurements. 

If p-values for an ANOVA test were below .05, the Tukey post-hoc test was used 

to determine which doses had significantly different averages for that particular 

measure. 

II. Summary of Results 

 For all three generations, females had greater average body weights compared 

to males. Females also tended to gain more weight throughout the duration of the 

study compared to males. For both genders, the F1 generation started out with 

much higher average body weights compared to the P and F2 generations. At 

about week 4, the three generations average bodyweights became more similar. 

Even with significant differences, body weight differences between dose group, 

within each generation, were very minimal. 

III. F0 Generation Results (Week 6 through Week 18) 

 For both genders in the P generation, the interaction was not significant. Week 

was a statistically significant factor for both genders, but dose was only 

significant for males. In general, as time increased, body weights increased for 

both genders. For males, Tukey post-hoc tests revealed that dose 125 had a 

significantly lower average body weight compared to the control group, dose 30, 
5 

and dose 60. In addition, dose 250 had a significantly lower body weight 

compared to dose 30 and dose 60. 

IV. F1 Generation Results (Week 2 to Week 7) 

 Prior to full onset of the study, additional information was collected on the 

chicks, but it was limited and sporadic. Data was collected for the tarsus length, 

the length from the tip of the beak to the culmen, as well as the full body weight. 

Minimal variation in tarsus and culmen averages occurred between the doses 

within each measurement day between week 2 and week 7. The only statistically 

significant difference occurred in the tarsus average on 7/13/2009. The control 

group had a significantly greater average compared to dose 60. In general, tarsus 

and culmen averages were so similar that no obvious patterns between dose 

groups could be detected. Significant differences in average body weight 

occurred more frequently than the tarus and culmen averages. However, the 

average body weight for the doses within each week were very similar and even 

the significant differences were minimal in magnitude. 

Factor P‐value Dose Week 6 Week 7 Week 8 Week 9 Week 10 Week 11 Week 12 Week 13 Week 14 Week 15 Week 16 Week 17 Week 18 Overall

0 105.1 113.9 124.7 130.0 129.8 132.1 131.5 136.3 137.4 138.2 134.1 141.5 138.0 130.2

30 108.9 120.5 125.5 125.7 129.7 133.3 136.4 138.2 138.2 139.5 139.7 140.4 142.3 132.2

60 103.7 115.2 125.3 127.6 130.5 133.7 131.5 134.6 136.9 137.0 138.3 140.1 142.9 130.6

125 104.3 116.6 126.7 129.5 131.9 131.5 135.9 135.9 134.0 137.7 140.7 138.6 141.6 131.1

250 105.1 117.6 125.4 130.3 133.7 132.3 134.3 135.0 141.9 139.4 133.9 140.7 143.0 131.7

Overall 105.4 116.7 125.5 128.6 131.1 132.5 133.9 136.0 137.7 138.3 137.3 140.3 141.5

F0Generation Female Body Weights

Significance Tests Estimated Marginal Means

Interaction ‐ 

dose by week 0.917

Dose   0.161

* Tukey post‐hoc tests indicate that weekly averages are signi ficantly different from each other.

Week 0.000*

Factor P‐value Dose Week 6 Week 7 Week 8 Week 9 Week 10 Week 11 Week 12 Week 13 Week 14 Week 15 Week 16 Week 17 Week 18 Overall

0 98.2 103.6 105.2 107.6 107.8 109.5 108.0 109.2 110.3 111.5 111.9 113.1 115.4 108.5

30 99.5 104.8 106.4 106.7 107.8 110.2 107.9 111.3 112.7 112.0 111.9 113.6 114.8 109.2

60 99.2 103.9 105.6 107.3 109.2 109.9 107.9 108.6 112.0 111.1 113.8 112.4 114.3 108.8

125 100.0 103.1 103.2 104.0 105.0 106.6 104.4 105.8 102.3 109.5 111.0 110.3 111.1 105.8

250 98.0 1010.9 103.5 105.4 106.8 108.0 106.6 108.3 108.6 108.8 110.4 111.5 113.9 107.0

Overall 99.0 103.4 104.8 106.2 107.3 108.8 106.9 108.6 109.2 110.6 111.8 112.2 113.9

* Tukey post‐hoc tests indicate that dose 125 had signi ficant lower body weight compared to the control group, dose 30, and dose 60.  Dose 250 had a significantly lower body weight compared to dose 30 and dose

60.

** Tukey post‐hoc tests indicate that weekly averages are signi ficantly di fferent from each other.

F0 Generation Male Body Weights

Significance Tests Estimated Marginal Means

Interaction ‐ 

dose by week 0.999

Dose   0.000*

Week 0.000**
6 

V. F1 Generation Results (Week 8 through Week 22) 

 The dose by week interaction was statistically significant for the females in the 

F1 generation. Dose 250 generally has the highest body weight average while the 

control group and dose 60 had the lowest body weight averages. 

 The dose by week interaction was not significant for the males; however, week 

and dose were highly significant. As week increased, body weight generally 

increased. Tukey post-hoc tests indicated that the control group’s average body 

weight was significantly lower than the other four dose groups. Additionally, 

dose 30 had a significantly lower body weight compared to dose 60. 

Week 2 Week 3 Week 4

Female Male Female Male Female Male

p‐value = .13 p‐value = .00 N/A* p‐value = .00 p‐value = .00 p‐value = .30 p‐value = .58 p‐value = .35 p‐value = .01

0 13.5 35.0 54.3 78.2 75.5 97.3 91.1 113.9 94.2

30 14.2 36.9 80.0 76.4 75.0 93.1 95.6 110.3 102.9

60 15.8 26.7 51.3 66.9 80.3 100.1 93.1 118.5 98.9

125 18.4 34.8 58.9 79.2 67.9 99.4 95.3 115.5 101.5

250 18.3 35.2 58.0 80.7 76.4 102.1 93.2 113.4 103.6

p‐value = .33 p‐value = .00 p‐value = .63 p‐value = .61 p‐value = .73 p‐value = .17 p‐value = .60 p‐value = .22 p‐value = .65

0 18.1 22.6 20.4 32.7 32.1 33.3 33.1 33.6 33.1

30 19.0 21.3 21.7 32.8 32.1 33.3 32.7 33.4 32.7

60 19.3 19.0 21.4 32.4 32.5 33.3 33.0 33.4 32.8

125 19.9 20.1 21.3 33.3 32.0 33.3 33.0 33.2 33.0

250 19.3 20.5 20.6 32.5 32.1 34.3 33.3 34.1 33.2

p‐value = .14 p‐value = .09 p‐value = .53 p‐value = .96 p‐value = .52 p‐value = .05 p‐value = .41 p‐value = .48 p‐value = .11

0 3.7 6.0 7.4 6.8 6.8 7.6 7.2 7.4 6.9

30 3.9 6.5 7.1 6.9 6.7 7.7 7.1 7.2 7.2

60 3.8 6.3 7.5 6.9 7.0 7.4 7.0 7.4 6.9

125 4.1 6.3 7.4 6.9 6.8 7.2 7.3 7.4 7.4

250 4.1 5.8 7.0 6.8 6.9 7.5 7.2 7.1 7.4

* Dose 30 only had 1 observa tion.  Dose 30 va ria nce was could not be calcula ted s o the ANOVA tes t was unable to be executed.

Dose

F1 Generation Tarsus, Culmen, and Body Weight Averages

Tarsus

Culmen

Week 5 Week 6 Week 7

Body Weight

Factor P‐value Dose Week 8 Week 9 Week 10 Week 11 Week 12 Week 13 Week 14 Week 15 Week 16 Week 17 Week 18 Week 19 Week 20 Week 21 Week 22 Overall

0 118.4 126.5 128.5 127.5 130.0 131.3 130.1 132.1 135.9 134.7 134.3 134.1 132.8 135.3 127.2 130.6

30 124.2 129.0 136.3 134.9 135.1 136.2 136.6 138.1 136.3 140.9 138.3 140.1 138.8 142.9 143.6 136.7

60 138.2 126.8 131.9 130.7 131.2 131.9 126.8 131.0 131.0 139.6 133.3 125.0 129.2 133.2 135.8 131.7

125 126.5 132.5 133.7 135.1 134.8 136.0 136.8 131.8 137.0 139.3 138.7 142.0 141.8 136.9 138.9 136.1

250 130.6 132.6 136.1 137.1 133.6 138.5 139.1 139.3 142.4 142.4 139.3 141.5 143.2 144.6 143.1 138.9

Overall 127.5 129.4 133.3 133.0 132.9 134.8 133.9 134.5 136.5 139.4 136.8 136.5 137.1 138.6 137.7

* Statistically signi ficant interaction indicated that body weight change from week to week was signi ficantly di fferent between the five dose groups.

Significance Tests Estimated Marginal Means

Interaction ‐ 

dose by week 0.005*

Dose   0.000

Week 0.000

F1 Generation Female Body Weights
7 

VI. F2 Generation Results (Week 0 through Week 4) 

Prior to full onset of the study, additional information was collected on the 

chicks, but it was limited and sporadic. Data was collected for the tarsus length, 

the length from the tip of the beak to the culmen, as well as the full body weight. 

No significant differences between doses occur for any of the body weight, tarsus, 

or culmen data collected in the F2 generation from week 0 to week 4. Negligible 

differences in averages occurred between doses, but there was no visible pattern 

between doses. 

Factor P‐value Dose Week 8 Week 9 Week 10 Week 11 Week 12 Week 13 Week 14 Week 15 Week 16 Week 17 Week 18 Week 19 Week 20 Week 21 Week 22 Overall

0 101.9 103.7 105.1 105.1 106.5 105.9 104.6 106.8 111.7 108.3 109.0 112.2 108.3 111.9 108.2 107.3

30 103.8 107.4 108.7 107.9 109.2 107.0 107.0 111.8 111.3 111.1 112.0 112.2 112.6 116.1 114.5 110.1

60 106.2 108.8 110.8 110.4 111.5 111.3 110.0 111.6 113.9 114.5 115.5 109.9 112.1 117.6 116.7 112.0

125 107.4 109.1 109.9 109.3 110.7 109.7 108.9 114.4 111.2 111.8 112.5 112.2 112.7 114.5 115.9 111.3

250 104.8 107.7 108.1 107.6 108.4 108.3 110.2 109.5 111.2 111.7 112.1 112.5 113.3 115.5 115.3 110.4

Overall 104.8 107.3 108.5 108.0 109.2 108.4 108.1 110.6 111.8 111.5 112.2 111.8 111.8 115.1 114.1

* Tukey post‐hoc tests indicated that the control group's body weight was significantly lower than the other four dose groups.  Dose 30 also ahd a significantly lower body weight compared to dose 60.

** Tukey post‐hoc tests indicate that weekly averages are signi ficantly di fferent from each other.

Estimated Marginal Means

Dose   0.000*

Significance Tests

Interaction ‐ 

dose by week 0.996

Week 0.000**

F1 Generation Male Body Weights

Dose Week 1 Week 2 ‐ A Week 2 ‐ B Week 3 ‐ A Week 3 ‐ B Week 4 ‐ A Week 4 ‐ B Week 5 ‐ A Week 5 ‐ B

p‐value = .91 p‐value = .94 p‐value = .47 p‐value = .57 p‐value = .95 p‐value = .77 p‐value = .83 p‐value = .65 p‐value = .78

0 7.6 16.2 24.8 34.6 46.9 57.0 70.3 78.7 81.8

30 7.2 16.9 25.7 36.2 47.7 60.3 72.3 80.6 84.0

60 7.7 17.2 26.6 37.4 48.2 59.6 72.0 79.2 83.2

125 7.7 17.1 24.6 36.8 47.7 60.3 73.1 82.5 84.6

250 7.6 16.3 23.9 36.0 46.6 58.4 74.0 82.8 86.0

p‐value = .80 p‐value = .26 p‐value = .28 p‐value = .31 p‐value = .14 p‐value = .39 p‐value = .16 p‐value = .26 p‐value = .74

0 15.6 17.8 20.9 23.0 27.2 28.0 30.3 31.0 32.7

30 15.8 19.4 21.4 23.4 26.9 29.7 31.5 31.8 32.8

60 16.1 19.1 21.3 24.1 28.3 29.7 31.7 31.9 32.3

125 15.8 19.8 19.0 23.1 27.1 28.6 31.8 32.8 32.3

250 16.0 18.9 20.1 22.4 26.0 28.5 31.5 32.4 33.1

p‐value = .37 p‐value = .70 p‐value = .43 p‐value = .76 p‐value = .57 p‐value = .56 p‐value = .26 p‐value = .79 p‐value = .82

0 3.8 4.7 5.6 6.3 6.7 7.3 7.4 7.9 7.8

30 3.7 4.7 5.6 6.5 6.6 7.4 7.8 8.0 7.7

60 3.9 4.9 5.7 6.6 5.8 7.7 7.5 8.1 7.9

125 3.7 4.9 5.9 6.3 5.9 7.3 7.4 7.9 7.8

250 4.0 4.8 5.5 6.6 6.6 7.4 7.2 7.9 7.7

Body Weight

Tarsus

Culmen

F2Generation Tarsus, Culmen, and Body Weight Averages
8 

VII. F2 Generation Results Week 1 through Week 14 

 Week and dose were statistically significant or both genders in the F2 

generation, and the interaction term was not. As week increased, average body 

weight for both genders steadily increased. In females, dose 60 had a 

significantly lower body weight average compared to dose 30 and dose 125. For 

males, the control group had a significantly lower body weight compared to the 

other four dose groups. Furthermore, in males, dose group 125 had a significantly 

higher body weight compared to the other four dose groups. 

 

Factor P‐value Dose Week 6 Week 7 Week 8 Week 9 Week 10 Week 11 Week 12 Week 13 Week 14 Week 15 Week 16 Week 17 Week 18 Week 19 Overall

0 100.9 108.8 124.3 134.5 133.5 132.3 137.8 137.9 140.3 141.6 138.3 138.3 141.1 141.8 132.2

30 106.7 116.0 124.2 130.5 133.9 135.6 137.6 138.5 139.7 143.2 139.8 140.0 140.2 142.1 133.4

60 102.1 108.2 124.5 131.5 131.6 132.5 135.9 138.3 138.5 140.0 136.5 136.5 138.6 137.2 130.9

125 103.5 112.6 125.2 129.6 136.1 139.3 139.7 140.1 140.4 142.4 139.6 139.6 142.2 145.0 133.9

250 108.1 115.7 125.8 128.1 131.6 133.4 135.9 137.7 139.3 142.0 140.0 140.1 140.2 140.6 132.7

Overall 104.2 112.2 124.8 130.8 133.3 134.6 137.5 138.5 139.6 141.8 138.8 138.9 140.4 141.3

F2 Generation Female Body Weights

Dose   0.003*

Week 0.000**

*  Tukey post‐hoc tests indicated that dose 60 had a significantly lower body weight compared to dose 30 and dose 125.

** Tukey post‐hoc tests indicate that weekly averages are signi ficantly different from each other.

Significance Tests

Interaction ‐ 

dose by week 0.948

Estimated Marginal Means

Factor P‐value Dose Week 6 Week 7 Week 8 Week 9 Week 10 Week 11 Week 12 Week 13 Week 14 Week 15 Week 16 Week 17 Week 18 Overall

0 94.3 98.1 103.4 104.4 104.7 106.1 108.5 108.5 108.7 109.4 109.8 110.1 110.5 105.9

30 100.4 104.2 107.3 109.4 109.5 110.7 112.6 112.7 113.4 114.1 114.1 115.6 114.4 110.6

60 97.6 101.9 106.0 108.0 108.7 110.0 110.8 112.0 111.4 112.9 112.9 112.8 113.2 109.1

125 102.8 106.9 109.7 111.7 113.4 113.6 115.1 115.6 114.3 115.1 114.3 114.7 114.7 112.4

250 101.0 103.6 104.1 107.2 108.6 110.2 111.9 113.0 111.2 112.8 113.3 113.5 113.6 109.5

Overall 99.2 102.9 106.1 108.1 109.0 110.1 111.8 112.3 111.8 112.8 112.9 113.3 113.2

** Tukey post‐hoc tests indicate that weekly averages are signi ficantly di fferent from each other.

* Tukey post‐hoc tests indicate that the control group had a significantly lower body weight compared to the other four dose groups.  Dose 125 had a significantly higher body weight compared to the four other dose

groups.

Interaction ‐ 

dose by week 1.000

Dose   0.000*

Week 0.000**

F2 Generation Male Body Weights

Significance Tests Estimated Marginal Means
9 

FEED CONSUMPTION 

I. Analysis Methods 

A repeated measured Linear Mixed Model was run to determine if there was a 

significant difference in food consumption between the five dose groups. A 

repeated measures model was used to test the dose and week as well as their 

interaction, while accounting for the variation in the data from week to week. 

Dose, week, and interaction were determined to be significant if the p-value for 

the Mixed Model test was less than .05. Estimated marginal means were 

calculated for both factors and examined for differences using a Tukey post-hoc 

test, if the p-value for that factor was less than .05. If the interaction was 

statistically significant, the interaction was investigated via marginal means. The 

significance results of the primary factors are considered to be uninformative and 

can be ignored. Also, the food consumption calculations that were incorrectly 

recorded as negative were excluded from the analysis since it was impossible for 

the birds to eat more feed than was originally in the container. 

II. Summary of Results 

Males and females typically started out with equal food consumption 

averages for the first weeks of data collection, however female food consumption 

increased extensively more throughout the unpaired weeks than males. No dose 

group consistently had higher food consumption averages for either the paired or 

unpaired weeks. Food consumption averages, overall, generally increased from 

one week to the next, however, certain weeks throughout all generations had very 

low or very high averages, making one question how valid the food consumption 

measurements really were. 

III. F0 Generation Results (Week 5 through Week 10 – unpaired genders) 

 In the P generation of females, the interaction was statistically significant. All 

doses increased from one week to the next; however, each dose tended to have 

higher rate of increase at different weeks. Dose 125 and dose 250’s increase in 

food consumption during week 8 through 10 were much less than the other dose 

groups. 

 For males, the dose by week interaction was not significant, but week and dose 

were. Food consumption was lowest at week one, but generally was steady after 

that. The control group had a significantly greater food consumption compared to 

all the other dose groups, which had similar food consumption averages. 
10 

IV. F0 Generation Results (Week 11 through Week 18 – paired genders) 

 From week 11 to week 18, the interaction was not statistically significant, 

however dose and week were. From week 11 to week 12, food consumption 

drastically increased. From week 12 to week 17, food consumption was fairly 

consistent until week 18, in which food consumption severely dropped. Dose 30 

and dose 125’s food consumption averages were significantly greater than the 

control group and dose 60 averages. 

V. F1 Generation Results (Week 7 through Week 12 – unpaired genders) 

For the F1 generation, both genders had non-significant interaction, but 

significant differences by week. For females, dose was also statistically 

significant. Food consumption generally increased one week to the next for both 

Factor P‐value Dose Week 5 Week 6 Week 7 Week 8 Week 9 Week 10 Overall

0 113.1 114.9 138.7 154.6 166.7 199.8 148.0

30 132.0 134.8 146.1 146.0 156.8 182.5 149.7

60 117.6 121.9 154.3 154.0 154.3 196.5 149.8

125 123.1 141.1 146.7 187.0 191.2 187.2 162.7

250 125.4 145.6 147.8 175.5 175.4 186.7 159.6

Overall 122.2 131.6 146.7 163.6 168.9 190.5

Significance Tests

Interaction ‐ 

dose by week 0.003

Dose   0.000

F0 Generation Female Food Consumption (Weeks 5‐10)

Estimated Marginal Means

Week 0.000

Factor P‐value Dose Week 5 Week 6 Week 7 Week 8 Week 9 Week 10 Overall

0 107.4 121.2 131.7 123.1 121.4 136.9 123.6

30 109.5 117.3 122.0 107.4 106.8 118.3 113.5

60 113.5 113.2 121.3 115.9 116.1 118.8 116.5

125 112.9 111.0 112.1 118.6 109.7 116.4 113.4

250 106.6 111.4 114.9 117.9 114.5 121.1 114.4

Overall 110.0 114.8 120.4 116.6 113.7 122.2

Interaction ‐ 

dose by week

0.111

* ID 69 ‐ dose 30 datapoints removed for week 9 and week 10 due to low food consumption results.

Dose   0.000

Week 0.000

F0 Generation Male Food Consumption (Weeks 5‐10)

Significance Tests Estimated Marginal Means

Factor P‐value Dose Week 11 Week 12 Week 13 Week 14 Week 15 Week 16 Week 17 Week 18 Overall

0 154.0 290.9 305.4 306.1 282.1 293.6 289.1 113.7 254.4

30 160.4 325.7 324.9 326.5 323.6 302.4 316.2 126.7 275.8

60 159.0 300.9 279.2 330.5 292.1 289.9 295.9 125.1 259.1

125 163.4 320.2 320.1 323.6 317.9 313.6 315.7 127.7 275.3

250 161.8 299.5 291.0 305.5 292.0 292.4 318.6 127.1 261.0

Overall 159.7 307.5 304.1 318.4 301.5 298.4 307.1 124.1

Dose  

0.000

F0Generation Food Consumption (Week 11‐18)

Significance Tests Estimated Marginal Means

Interaction ‐ 

dose by week

0.640

0.000

Week
11 

genders. For females, dose group 250 had a significantly larger food consumption 

compared to all other doses. Even though significant differences did not occur 

between dose groups for males, dose 60 had the greatest food consumption 

average while dose 30 had the lowest food consumption average. 

VI. F1 Generation Results (Week 13 through Week 21 – paired genders) 

 For the paired birds in week 13 through week 21, week and dose were 

significant while their interaction was not. Tukey post-hoc results indicated that 

dose 250 and the control group had significantly greater food consumption 

averages compared to dose 125. Additionally, the control group had a 

significantly greater food consumption average compared to dose 30. Week 13 

and week 14’s averages were well below the averages through the rest of the F1 

generation. 

Factor P‐value Dose Week 7 Week 8 Week 11 Week 12 Overall

0 74.6 126.1 150.1 196.8 131.3

30 78.4 123.4 151.5 190.5 129.9

60 102.9 131.6 147.0 199.8 137.4

125 89.5 124.7 143.4 202.1 133.8

250 103.3 145.5 163.1 215.1 149.5

Overall 89.7 130.3 151.0 200.8

F1 Generation Female Food Consumption (Weeks 7‐12)

Significance Tests Estimated Marginal Means

Week 0.000

* Id 119, 12, 299, 163, and 163 datapoints removed for week 7 due to low food consumption results.

Interaction ‐ 

dose by week 0.069

Dose   0.000

Factor P‐value Dose Week 7 Week 8 Week 9 Week 11 Week 12 Overall

0 79.4 90.1 97.9 95.6 124.1 97.6

30 71.9 91.6 95.1 93.3 117.9 94.0

60 68.5 97.5 102.5 102.7 124.1 99.1

125 70.5 95.5 89.6 102.4 126.1 97.1

250 74.7 95.8 96.0 100.5 120.3 97.7

Overall 73.0 94.5 96.2 99.1 122.5

Interaction ‐ 

dose by week

0.157

Dose   0.188

Week 0.000

Significance Tests

F1Generation Male Food Consumption (Weeks 7‐12)

Estimated Marginal Means
12 

VII. F2 Generation Results (Week 6 through Week 18 – unpaired genders) 

 The interaction was statistically significant for both genders in the F2 

generation. For females, the interaction was significant because of the large drop 

in food consumption for dose 60 and dose 30 in week 16. The control group for 

females did have the highest overall food consumption averages, but did not have 

the highest average throughout all thirteen weeks. 

 For males, minimal differences in food consumption occurred between dose 

groups within each week, and no particular dose group that generally had greater 

or lower food consumption averages compared to the other doses. The interaction 

was found significant due to the greater fluctuation in food consumption in the 

dose 250 group compared to the other doses. 

 Overall, both genders had similar food consumption results until week 10 

where females started consuming far more food. Variation between doses was 

much greater for females compared to males, and no particular dose constantly 

had a higher or lower food consumption compared to the others for either gender. 

 

Factor P‐value Dose Week 13 Week 14 Week 15 Week 16 Week 17 Week 18 Week 19 Week 20 Week 21 Overall

0 179.3 130.9 346.4 293.4 315.3 309.1 286.7 327.8 257.7 271.9

30 161.7 142.6 338.3 263.2 291.0 298.4 278.4 270.8 248.9 254.8

60 173.4 138.4 329.1 266.5 290.9 286.1 268.1 331.4 241.0 258.3

125 161.0 130.9 323.3 246.9 297.9 294.5 294.3 279.7 243.6 252.5

250 172.8 131.8 336.8 281.8 301.5 308.1 304.2 316.7 273.4 269.7

Overall 169.6 134.9 334.7 270.4 299.3 299.2 286.3 305.3 252.9

Week 0.000

0.001

Estimated Marginal Means

F1 Generation Food Consumption (Week 13‐21)

Significance Tests

Interaction ‐ 

dose by week 0.559

Dose  

Factor P‐value Dose Week 6 Week 7 Week 8 Week 9 Week 10 Week 11 Week 12 Week 13 Week 14 Week 15 Week 16 Week 17 Week 18 Overall

0 91.1 114.7 138.7 98.8 130.7 195.8 219.1 243.9 193.7 231.8 214.3 244.3 243.2 181.8

30 103.4 120.9 125.0 112.4 134.6 183.6 197.7 216.3 202.2 212.5 111.8 217.8 215.5 165.7

60 73.5 108.8 128.3 91.1 136.4 193.9 207.0 236.3 200.7 213.2 107.9 235.1 234.7 166.7

125 94.5 116.0 125.3 91.7 139.3 169.6 178.0 222.6 183.3 198.7 195.2 207.3 223.4 165.0

250 84.5 106.8 123.9 100.8 127.4 187.3 202.5 249.1 193.9 202.0 179.0 222.2 227.3 169.7

Overall 90.0 113.4 128.2 98.9 133.7 186.2 200.8 233.6 194.7 211.6 161.6 225.3 228.8

* Id 305 (week 9), 215 (week 9), 86 (week 16), 73 (week 16), 84 (week 16) datapoints removed due to inaccurate food consumption results

Interaction ‐ 

dose by week 0.000

Dose   0.000

F2 Generation Female Food Consumption (Week 6‐18)

Estimated Marginal Means

Week 0.000

Significance Tests

Factor P‐value Dose Week 6 Week 7 Week 8 Week 9 Week 10 Week 11 Week 12 Week 13 Week 14 Week 15 Week 16 Week 17 Week 18 Overall

0 97.8 120.8 122.0 114.6 104.6 121.3 120.9 196.0 119.1 133.7 118.2 116.2 118.1 123.3

30 98.9 115.4 119.6 122.4 107.7 113.6 110.4 200.1 122.8 129.9 112.6 108.5 116.0 121.3

60 89.2 121.7 127.2 123.9 115.2 122.1 130.0 204.1 123.9 127.6 114.0 119.8 124.2 126.4

125 94.2 125.0 129.3 122.0 122.5 120.1 116.1 193.8 119.1 126.0 106.6 113.5 127.9 124.3

250 96.6 116.8 117.3 103.9 132.5 122.1 124.5 198.1 122.3 124.0 109.2 112.2 121.3 123.1

Overall 95.3 119.9 123.1 117.4 116.5 119.8 120.3 198.4 121.4 128.2 112.1 114.0 121.5

* ID 193 datapoint removed from week 7 due to inaccurate food consumption result.

F2Generation Male Food Consumption (Week 6‐18)

Estimated Marginal Means

Dose   0.077

Week 0.000

Significance Tests

Interaction ‐ 

dose by week 0.003
13 

CLOACA GLAND (CG) AREA 

I. Analysis Methods 

 A General Linear Model – Repeated Measures (GLMREP) test was used to 

determine if the CG area was statistically difference between the five dose groups, 

while accounting for the variation throughout weeks. P-values were calculated 

for the dose and week as well as their interaction. Dose, week, and interaction 

were determined to be significant if the p-value for the GLM was less than .05. 

Estimated marginal means were calculated for both factors and examined for 

differences using a Tukey post-hoc test, if the p-value for that factor was less than 

.05. If the interaction was statistically significant, the interaction was investigated 

via marginal means. The significance results of the primary factors are 

considered to be uninformative and can be ignored. 

II. Summary of Results 

 CG area was the largest overall for the F1 generation. Additionally, in all three 

generations, CG area, not surprisingly, increased from one week to the next. 

Statistically significant differences occurred between doses in the P and F1 

generations, but there was no distinct pattern between an increased dose 

concentration and either an increase or decrease in CG area. In the P and F1 

generations, however, the control group did have the largest CG area. 

III. F0 Generation Results 

 The dose by week interaction was not statistically significant, meaning that 

from week 6 to week 7, the five doses increased CG area at the same rate. 

However, dose and week were both statistically significant. Week 6’s CG area 

was significantly larger than week 7’s. The control group has the largest average 

CG area for both weeks, and post-hoc analysis revealed that the control group had 

a significantly greater CG area compared to dose 60. 
14 

IV. F1 Generation Results 

 As in the F0 generation, dose and week were statistically significant factors, 

while the interaction was not. Week 7’s CG area was significantly larger than 

week 6’s CG area. The control group has the largest average CG area, and posthoc analysis revealed that the control group had a significantly greater CG area 

compared to dose 30. Dose 60 had next largest CG area, just slightly smaller than 

the control group. 

V. F2 Generation Results 

 In the F2 generation, the dose by week interaction and dose were not 

statistically significant, however week was. The CG area’s increases from week 6 

to week 7 and from week 7 to week 8 were both statistically significant. Overall, 

dose 30 had the largest and dose 250 had the smallest CG area average. 

Factor P‐value Dose Week 6 Week 7 Overall

0 113.4 164.4 138.9

30 75.4 118.6 97.0

60 70.0 114.2 92.1

125 97.7 165.0 131.4

250 84.1 136.7 110.4

Overall 88.1 169.8

Estimated Marginal Means

F0Generation Cloaca

Interaction ‐ 

dose by week

Dose

Week

0.230

0.013*

0.000**

Significance Tests

* Tukey post‐hoc tests indicate that the control group had a significantly greater CG

area compared to dose 60.

** Week 7 had significantly greater CG area compared to week 6.

Factor P‐value Dose Week 6 Week 7 Overall

0 139.7 189.3 164.5

30 89.2 157.0 123.1

60 150.7 177.2 163.9

125 109.5 173.5 141.5

250 119.6 160.8 140.2

Overall 121.7 171.5

Interaction ‐ 

dose by week

0.588

Dose 0.017*

Week 0.000**

** Week 7 had a signi ficantly greater CG area average compared to week 6.

* Tukey post‐hoc tests indicated that the control group had a signi ficantly greater

CG area compared to dose 30.

F1 Generation Cloaca

Significance Tests Estimated Marginal Means
15 

 

Factor P‐value Dose Week 6 Week 7 Week 8 Overall

0 104.0 151.5 205.3 153.6

30 100.4 160.8 218.3 159.8

60 76.8 152.8 210.0 146.5

125 97.4 148.6 225.8 157.3

250 74.7 148.9 201.7 141.8

Overall 90.6 152.5 212.2

* All weeks were signi ficantly different from each other.

Significance Tests Estimated Marginal Means

F2 Generation Cloaca

Week 0.000*

Interaction ‐ 

dose by week

0.785

Dose 0.905
16 

YOLK CONTENT 

I. Analysis Methods 

 A non-parametric Kruskal-Wallis test was used to test if there was a significant 

difference among doses in total endosulfans, I endosulfans, and II endosulfans 

present in the yolk. A Kruskal-Wallis test is the equivalent of the parametric 

ANOVA test, however fewer assumptions have to be met. Non-parametric 

models rank the dataset in terms of magnitude and test to see if there are 

differences in group medians/ranks rather than means. Equal variances of the 

groups is not necessary and ample sample size per group is not required. With 

unequal and small sample sizes per group, the non-parametric test was more 

applicable for the yolk content data than other statistical tests. 

II. P Generation Results 

Kruskal-wallis average ranks indicated that endosulfans (Total, I, II) 

increased as dose concentration increased. This was true for all three yolk content 

measures. The control group and dose 30 were significantly different from dose 

125 and dose 250 for all three measures. 

 

Dose Total Endosulfans I II

P‐value 0.001 0.002 0.001

0 0.00 0.01 0.00

30 0.12 0.03 0.12

60 0.39 0.07 0.36

125 0.66 0.09 0.64

250 1.50 0.23 1.49

P Generation Yolk Content Medians
17 

ONSET TO PUBERTY 

I. Analysis Methods 

 An Analysis of Variance (ANOVA) test was used to determine if onset to 

puberty averages for dose groups were significantly different. An F-statistic and 

corresponding p-value was calculated for each gender within each generation. If 

the p-value was below .05, then it was concluded that at least two dose groups had 

statistically different averages for onset to puberty. A Tukey post-hoc test was 

then used to find which specific dose groups were statistically different from one 

another. A Levene’s test was used to check to make sure the dose groups had 

equal variances, a major assumption that must be met in order to properly run an 

ANOVA model. If this assumption was violated (p-value for Levene’s test < 

.05), a non-parametric Kruskal-Wallis test was run instead of an ANOVA. 

II. Summary of Results 

 For all dose groups in all three generations, females onset to puberty was 

longer than males. Additionally for all dose groups in both genders (excluding 

dose 30 females and dose 250 males), onset to puberty decreased in length from 

the F0 to F1 generation and increased from the F1 to F2 generation. For females, 

over all three generations, there was no specific dose group that typically had the 

shortest or longest onset to puberty averages. However, for males, dose 30 and 

dose 250 generally had the longest onset to puberty out of all dose groups. 

Overall, minimal statistical evidence showed that there were statistically 

significant differences between dose groups for onset to puberty in either males or 

females. 

III. F0 Generation Results 

 No statistically significant differences occurred between dose groups for either 

gender within the F0 generation. Males had a p-value slightly below .05, but after 

accounting for multiple testing, there were no significant differences between 

dose groups. For males, dose 30 had the longest onset to puberty. 
18 

IV. F1 Generation Results 

 Statistically significant differences occurred between dose groups for the 

females in the F1 generation. Dose 30 had a significantly longer onset to puberty 

compared to the control group and dose 60. For the males, even though there 

were no significant differences, dose 30 and dose 250 had the longest averages 

and dose 60 had the shortest averages. 

V. F2 Generation Results 

 With p-values well above .05, statistically significant differences between dose 

groups were not present for either gender, comparing onset to puberty averages. 

For females, dose 125 had the longest average while the control group had the 

shortest averages. For males, dose 30 had the longest and dose 125 had the 

shortest average onset to puberty. 

Dose Average Std. Error Average Std. Error

0 57.2 1.68 44.7 0.47

30 56.1 1.81 46.8 0.67

60 56.6 1.70 45.9 0.64

125 56.3 1.05 44.7 0.51

250 56.3 1.44 45.8 0.59

P‐value = .991 P‐value = .044

Females Males

F0 Generation Onset to Puberty (days)

Dose Average Std. Error Average Std. Error

0 49.6 1.44 43.2 2.47

30 55.4 1.31 46.0 1.86

60 49.9 1.59 41.6 1.92

125 54.3 1.27 44.3 1.84

250 53.7 1.17 46.5 1.92

Females Males

P‐value = .007* P‐value = .405

* Tukey post‐hoc tests indicate that dose 30 has

signi ficantly longer onset to puberty compared to the

control group and dose 60.

F1Generation Onset to Puberty (days)
19 

 

Dose Average Std. Error Average Std. Error

0 53.2 1.94 48.1 1.32

30 54.4 2.18 48.7 1.46

60 55.7 1.72 48.1 1.26

125 57.3 1.81 47.1 1.23

250 55.3 2.66 48.1 1.24

F2 Generation Onset to Puberty (days)

Females Males

P‐value = .680 P‐value = .933
20 

ENDOSULFAN EFFECTS ON REPRODUCTIVE

BEHAVIORS

Mounting and Copulation Disruption

 This section of the report analyzes the sexual initiation behavior characteristics 

of male Japanese quails through three generations (F0, F1, and F2). Specifically, 

the number of mount attempts, time to first mount and first copulation, and 

number of copulations were analyzed. Each pair of quails was part of one of five 

dose groups: (control) 0 ppm, 30 ppm, 60 ppm, 125 ppm, and 250 ppm. Data was 

collected on the same pairs through three consecutive trials in each of the three 

generations. SPSS version 16.0 was used to analyze the behavior data. Statistical 

significance for this analysis was determined using an alpha (α) of .05 with a 95% 

confidence level. 

 
21 

NUMBER OF MOUNT ATTEMPTS 

I. Analysis Methods 

 A Generalized Linear Model (GzLM) using a poisson loglinear link function 

was used to determine if there was a statistically significant difference in the 

number of mounts attempted across the five dose levels. An omnibus likelihood 

ratio test in GzLM, which compares the fitted model (intercept plus dose) against 

the intercept only model, was used to determine if dose was a significant factor in 

predicting the number of mounts in 180 seconds. If the p-value for the likelihood 

test was less than .05, dose was a significant factor in predicting the number of 

mount attempts. Parameter estimates (Beta coefficients and Beta confidence 

intervals) were then examined to find where the significant differences occurred 

between the dose levels if the p-value for the omnibus test was less than .05. In 

the GzLM procedure, a reference group was needed for the other dose levels to be 

tested against for a significant difference. This was because we fit a categorical 

predictor into a linear model. The reference group for this analysis was the 

‘dose=0’ group which is referred to as the control group throughout the report. 

An exponentiated β coefficient of 1.00 meant that the dosed group was 1.00 times 

as likely to have a mount compared to the control group, or in other words, the 

two groups had an equal chance of having a mount. If the confidence interval 

contained 1.00, there was no statistical difference between the two groups and 

vice versa. 

II. Summary of Results 

 For all dose groups, the number of mount attempts was much higher for the F0 

generation compared to the latter two generations. Statistically significant 

differences between dose groups occurred only in the P generation, mainly due to 

the high number of mounts attempted for dose 125 in the first two trials. 

III. F0 Generation Results 

 Significant differences between dose groups occurred in the first two trials for 

the F0 generation. In trial 1, the control group and dose 125 had significantly 

greater number of mount attempts compared to dose 30, dose 60, and dose 250. 

In trial 2, dose 125 had a significantly greater number of mount attempts 

compared to the control group, dose 30 and dose 60. Overall, dose 125 

consistently had one of the highest estimated number of mount attempts. The 

control group was also the only dose group to decline in attempted mounts from 

both trial 1 to trial 2 and trial 2 to trial 3. 
22 

IV. F1 Generation Results 

 Likelihood ratio p-values, which were well above .05, indicated that there were 

no statistically significant differences between doses within any of the three trials, 

comparing the number of mount attempts. From trial to trial, the number of 

mount attempts per dose group slightly increased, with the exception of the 

control group’s decline from trial 2 to trial 3. In trials 1 and 2, the control group 

had the highest estimated number of attempted mounts, but in trial 3, the control 

group’s estimated number of mount attempts was the lowest out of all five dose 

groups. 

V. F2 Generation Results 

 Likelihood ratio p-values, which were well above .05, indicated that there were 

no statistically significant differences between doses within any of the three trials, 

comparing the number of mount attempts. However, in all three trials, the control 

group had the lowest number of estimated mount attmpts, and dose 30 had the 

highest number of estimated mount attempts. Overall, variation from trial to trial 

was very minimal, with the sharpest increases and decreases, from trial to trial, 

occurring in the dose 30 and dose 60 group, respectively. 

Dose

Average number

of Mounts

95% β Confidence

Interval* P‐value Average number

of Mounts

95% β Confidence

Interval* P‐value Average number

of Mounts

95% β Confidence

Interval* P‐value

0 6.60 4.95 4.40

30 4.60 .53 ‐ .91 0.01 4.35 .66 ‐ 1.18 0.39 5.00 .81 ‐ 1.44 0.38

60 4.20 .48 ‐ .84 0.00 4.75 .72 ‐ 1.28 0.78 5.35 .89 ‐ 1.58 0.17

125 6.55 .78 ‐ 1.26 0.95 6.90 1.07 ‐ 1.81 0.01 5.21 .92 ‐ 1.61 0.25

250 4.40 .51 ‐ .87 0.00 5.55 .85 ‐ 1.48 0.41 4.75 .81 ‐ 1.44 0.61

* Confidence Intervals reflect comparison to control group (ppm = 0).  Ppm levels with a confidence interval not including 1.00 is considered to be significantly different

F0Generation ‐ Number of Mounts

Trial 1

Omnibus Likelihood ratio p‐value = .000

Trial 2

Omnibus Likelihood ratio p‐value = .006

Trial 3

Omnibus Likelihood ratio p‐value = .676

Dose

Average number

of Mounts

95% β Confidence

Interval* P‐value Average number

of Mounts

95% β Confidence

Interval* P‐value Average number

of Mounts

95% β Confidence

Interval* P‐value

0 2.45 2.80 2.20

30 1.90 .51  ‐  1.19 0.24 2.20 .53 ‐ 1.17 0.23 2.21 .66 ‐ 1.53 0.98

60 1.95 .52  ‐  1.21 0.29 2.70 .66 ‐ 1.40 0.85 2.75 .84 ‐ 1.86 0.27

125 2.00 .54  ‐  1.24 0.34 1.95 .46 ‐ 1.05 0.08 2.50 .76 ‐ 1.70 0.54

250 2.05 .55  ‐  1.27 0.40 2.32 .56 ‐ 1.23 0.35 2.65 .79 ‐ 1.82 0.38

* Confidence Intervals reflect comparison to control group (ppm = 0).  Ppm levels with a confidence interval not including 1.00 is considered to be significantly different

Trial 1 Trial 2 Trial 3

Omnibus Likelihood ratio p ‐ value = .773 Omnibus Likelihood ratio p ‐ value = .381 Omnibus Likelihood ratio p ‐ value = .740

F1Generation  ‐  Number of Mounts
23 

TIME UNTIL FIRST MOUNT ATTEMPT 

I. Analysis Methods 

 Kaplan-Meier survival analysis was used to determine if there was a significant 

difference between the five dose levels in terms of the amount of time it took the 

birds for their first mount attempt. If their time was recorded as 180 seconds, this 

meant that the quail did not make a mount attempt and that observation was 

considered censored. A log rank statistic was computed to test whether the 

different dose groups had statistically different times until the first mount attempt. 

If the p-value associated with the log rank statistic was less than .05, it was 

concluded that the times between at least two of the dose groups were not 

statistically equal. To find which dose groups did not have equal times, estimated 

confidence intervals for the time to attempted mount were compared. If two dose 

groups had confidence intervals that did not overlap, these two doses were 

considered to have statistically different times at the 95% confidence level. 

II. Summary of Results 

Trial 2

Dose

Average number

of Mounts

95% β Confidence

Interval* P‐value Average number

of Mounts

95% β Confidence

Interval* P‐value Average number

of Mounts

95% β Confidence

Interval* P‐value

0 1.95 1.90 1.65

30 2.25 .75 ‐ 1.77 0.51 2.55 .88 ‐ 2.04 0.17 2.21 .85 ‐ 2.11 0.21

60 1.95 .64 ‐ 1.56 1.00 1.50 .49 ‐ 1.27 0.33 2.10 .81 ‐ 2.01 0.30

125 2.20 .73 ‐ 1.74 0.58 2.05 .69 ‐ 1.68 0.74 1.80 .68 ‐ 1.75 0.72

250 2.05 .68 ‐ 1.63 0.82 2.10 .71 ‐ 1.71 0.66 2.15 .83 ‐ 2.05 0.25

* Confidence Intervals reflect comparison to control group (ppm = 0).  Ppm levels with a confidence interval not including 1.00 is considered to be significantly different

F2Generation ‐ Number of Mounts

Trial 1

Omnibus Likelihood ratio p‐value = .945

Trial 3

Omnibus Likelihood ratio p‐value = .221 Omnibus Likelihood ratio p‐value = .659
24 

In general, through all three generations, dose 30 and dose 250 had the 

shortest time until first mount attempt. Additionally, almost all dose groups had 

the longest time to first attempted mount in trial 1. This was true for all three 

generations. In the first two generations, minimal, non-statistically significant 

differences occurred between dose groups within each trial. However, greater 

variation between dose groups occurred in the F2 generation. In the F2 

generation, the control group and dose 60 generally had the longest time until the 

first mount attempt, and dose 30 had, by far, the shortest time until first mount 

attempt. 

III. F0 Generation Results 

 Log rank p-values indicated that there were no statistically significant 

differences between any doses within any of the three trials, comparing the time 

to first mount attempt. Time until first mount attempt decreased greatly from trial 

1 to trial 2, however, only slightly decreased from trial 2 to trial 3. Dose group 

125 and dose 250 generally had the shortest estimated time until first mount 

attempt in all three trials. 

IV. F1 Generation Results 

 Log rank p-values, which were all well above .05, indicated that there were no 

statistically significant differences between any doses within any of the three 

trials, comparing the time to first mount attempt. Just like in the P generation, the 

estimated time until first attempted mount decreased from one trial to the next. 

Mount times were typically lowest for dose 30 and dose 250, and were generally 

the highest for the control group and dose 125. 

Dose

0 38.5 12.7 90% 21.9 11.8 90% 21.5 11.8 90%

30 58.1 14.6 80% 27.8 11.9 95% 14.7 8.6 95%

60 77.6 14.8 80% 28.6 10.0 95% 22.5 9.2 95%

125 26.5 9.3 95% 18.7 9.2 95% 15.7 9.0 95%

250 47.9 13.7 85% 16.5 6.3 100% 10.8 4.1 100%

F0 Generation ‐ Time to first Mount

Trial 1 Trial 2 Trial 3

Log rank p‐value = .069 Log rank p‐value = .830 Log rank p‐value = .688

Average time to

first mount

Standard

Error

Mounted

Pairs

Average time to

first mount

Standard

Error

Mounted

Pairs

Average time to

first mount

Standard

Error

Mounted

Pairs
25 

V. F2 Generation Results 

 Log rank p-values indicated that significant differences between doses 

occurred in all three trials. In each trial, the control group had a significantly 

longer time to mount attempt compared to dose 30. In trial 2, dose 60, dose 125, 

and dose 250 also had significantly longer time to first mount attempt compared 

to dose 30. Just like the two previous generations, time to first mount attempt 

typically decreased from one trial to the next. Dose 30 had a drastically smaller 

time to first mount attempt through all three generations compared to the other 

doses. Conversely, the control group had the longest time until first mount 

attempt for all three generations. 

 

Dose

0 59.4 14.8 85% 44.8 15.8 80% 37.9 15.2 85%

30 43.8 13.2 85% 27.2 11.8 90% 21.6 11.8 90%

60 60.1 13.2 90% 37.8 12.5 90% 10.6 2.9 100%

125 67.9 15.9 75% 37.4 12.0 95% 40.5 13.9 90%

250 44.3 11.9 90% 20.0 9.2 95% 15.8 9.5 94%

Mounted

Pairs

Average time to

first mount

Standard

Error

Mounted

Pairs

Average time to

first mount

Standard

Error

Mounted

Pairs

Average time to

first mount

Standard

Error

F1Generation ‐ Time to first Mount

Trial 1 Trial 2 Trial 3

Log rank p‐value = .795 Log rank p‐value = .790 Log rank p‐value = .444

Dose

0 84.9 18.2 60% 62.6 16.4 80% 47.9 15.3 80%

30 17.6 4.5 100% 3.4 0.5 100% 5.1 2.7 100%

60 57.7 16.9 80% 49.4 15.8 80% 22.8 9.3 95%

125 49.8 13.7 85% 26.9 11.1 95% 17.4 8.9 95%

250 50.2 13.7 85% 24.7 10.2 95% 17.1 9.0 95%

Mounted

Pairs

Average time to

first mount

Standard

Error

Mounted

Pairs

Average time to

first mount

Standard

Error

Mounted

Pairs

Average time to

first mount

Standard

Error

F2 Generation ‐ Time to first Mount

Trial 1 Trial 2 Trial 3

Log rank p‐value = .032 Log rank p‐value = .000 Log rank p‐value = .016
26 

NUMBER OF COPULATIONS 

I. Analysis Methods 

 A Generalized Linear Model (GzLM) using a poisson loglinear link function 

was used to determine if there was a statistically significant difference in the 

number of copulations (successful cloacal contact) across the five dose levels. An 

omnibus likelihood ratio test in GzLM, which compares the fitted model 

(intercept plus dose) against the intercept only model, was used to determine if 

dose was a significant factor in predicting the number of copulations in 180 

seconds. If the p-value for the likelihood test was less than .05, dose was a 

significant factor in predicting the number of copulations. Parameter estimates 

(Beta coefficients and Beta confidence intervals) and confidence intervals were 

then examined to find where the significant differences occurred between the dose 

levels if the p-value for the omnibus test was less than .05. In the GzLM 

procedure, a reference group was needed for the other dose levels to be tested 

against for a significant difference. This was because we fit a categorical 

predictor into a linear model. The reference group for this analysis was the 

‘dose=0’ group which is referred to as the control group throughout the report. 

An exponentiated β coefficient of 1.00 meant that the dosed group was 1.00 times 

as likely to have a copulation compared to the control group, or in other words, 

the two groups had an equal chance of having a copulation. If the confidence 

interval contained 1.00, there was no statistical difference between the two groups 

and vice versa. 

II. Summary of Results

 In each generation, particularly the F0 generation, the number of copulations 

increased from one trial to the next. Dose 60 had by far the lowest number of 

copulations in the F0 generation. For all three generations, dose 30 had one of, if 

not the highest estimated number of copulations of all dose groups. Very little 

statistical evidence shows that there were consistent differences between any of 

the dose groups for the number of copulations.

III. F0 Generation Results 

 Likelihood ratio p-values, which were well above .05, indicated that there were 

no statistically significant differences between any doses within any of the three 

trials, comparing the number of copulations. Estimated number of copulations for 

all dose groups within each trial were all below 1.00, however the number of 

copulations did increase from one trial to the next. Dose 60 had the lowest 
27 

number of copulations through all three trials, while the other four dose groups 

had almost equal counts. 

IV. F1 Generation Results 

 Likelihood ratio p-values, which were well above .05, indicated that there were 

no statistically significant differences between any doses within any of the three 

trials, comparing the number of copulations. Trial 3 had much higher copulation 

estimated compared to trial 1 and trial 2. Patterns between the dose groups were 

not apparent throughout the trials further indicating any differences between the 

dose groups were very minimal. 

V. F2 Generation Results 

 Statistically significant differences occurred in trial 1 of the F2 generation. 

The control group had a significantly lower estimated number of copulations 

compared to dose 30. Trends between trials or between doses were not apparent, 

emphasized by the large p-values in trial 2 and trial 3. 

Trial 2

Dose

Average number

of Copulations

95% β Confidence

Interval* P‐value Average number

of Copulations

95% β Confidence

Interval* P‐value Average number

of Copulations

95% β Confidence

Interval* P‐value

0 0.30 0.42 0.50

30 0.35 .39 ‐ 3.47 0.78 0.50 .49 ‐ 3.01 0.72 0.65 .57 ‐ 2.97 0.53

60 0.10 .07 ‐ 1.65 0.18 0.15 .09 ‐ 1.34 0.13 0.20 .13 ‐ 1.28 0.12

125 0.20 .19 ‐ 2.36 0.53 0.58 .55 ‐ 3.42 0.49 0.55 .47 ‐ 2.59 0.83

250 0.30 .32 ‐ 3.10 1.00 0.40 .36 ‐ 2.53 0.92 0.55 .47 ‐ 2.59 0.83

* Confidence Intervals reflect comparison to control group (ppm = 0).  Ppm levels with a confidence interval not including 1.00 is considered to be significantly different

Omnibus Likelihood ratio p‐value = .205 Omnibus Likelihood ratio p‐value = .226

F0Generation ‐ Number of Copulations

Trial 1

Omnibus Likelihood ratio p‐value = .458

Trial 3

Trial 1

Dose

Average number

of Copulations

95% β Confidence

Interval* P‐value Average number

of Copulations

95% β Confidence

Interval* P‐value Average number

of Copulations

95% β Confidence

Interval* P‐value

0 0.35 0.50 0.75

30 0.60 .68 ‐ 4.35 0.26 0.55 .47 ‐ 2.59 0.83 1.00 .68 ‐ 2.62 0.41

60 0.60 .68 ‐ 4.35 0.26 0.50 .42 ‐ 2.40 1.00 0.75 .49 ‐ 2.05 1.00

125 0.35 .35 ‐ 2.85 1.00 0.45 .37 ‐ 2.22 0.82 0.40 .23 ‐ 1.26 0.15

250 0.40 .41 ‐ 3.12 0.80 0.71 .61 ‐ 3.27 0.42 0.53 .31 ‐ 1.61 0.41

* Confidence Intervals reflect comparison to control group (ppm = 0).  Ppm levels with a confidence interval not including 1.00 is considered to be significantly different

Trial 3

Omnibus Likelihood ratio p‐value = .193

Trial 2

Omnibus Likelihood ratio p‐value = .581 Omnibus Likelihood ratio p‐value = .873

F1Generation ‐ Number of Copulations
28 

 

Trial 1 Trial 2

Dose

Average number

of Copulations

95% β Confidence

Interval* P‐value Average number

of Copulations

95% β Confidence

Interval* P‐value Average number

of Copulations

95% β Confidence

Interval* P‐value

0 0.35 0.60 0.65

30 1.10 1.34 ‐ 7.36 0.01 0.75 .59 ‐ 2.67 0.57 1.16 .90 ‐ 3.54 0.10

60 0.50 .54 ‐ 3.75 0.47 0.55 .40 ‐ 2.08 0.84 0.65 .46 ‐ 2.16 1.00

125 0.50 .54 ‐ 3.75 0.47 0.60 .45 ‐ 2.23 1.00 0.60 .42 ‐ 2.02 0.84

250 0.65 .74 ‐ 4.66 0.18 0.65 .49 ‐ 2.37 0.84 0.90 .68 ‐ 2.83 0.37

* Confidence Intervals reflect comparison to control group (ppm = 0).  Ppm levels with a confidence interval not including 1.00 is considered to be significantly different

F2Generation ‐ Number of Copulations

Omnibus Likelihood ratio p‐value = .043 Omnibus Likelihood ratio p‐value = .950

Trial 3

Omnibus Likelihood ratio p‐value = .268
29 

TIME UNTIL FIRST COPULATION 

I. Analysis Methods 

 Kaplan-Meier survival analysis was used to determine if there was a significant 

difference between the five dose levels in terms of the amount of time it took the 

birds to achieve their first copulation. If their time was recorded as 180 seconds, 

this meant that the quail did not copulate and that observation was considered 

censored. A log rank statistic was computed to test whether the different dose 

groups had statistically different times until the first copulation. If the p-value 

associated with the log rank statistic was less than .05, it was concluded that the 

times between at least two of the dose groups were not statistically equal. To find 

which dose groups did not have equal times, estimated confidence intervals for 

the time to copulate were compared. If two dose groups had confidence intervals 

that did not overlap, these two doses were considered to have statistically 

different times at the 95% confidence level. 

II. Summary of Results 

In all three generations, trial 1 had the longest time until first copulation. 

The variation between doses in the F2 generation was far greater than the 

variation in the P and F1 generation. Much of the F2 variation was due to the 

control group’s longer and dose 30’s significantly shorter times until copulation. 

Other than the large variation between doses in the F2 generation, the dose groups 

tended to follow the same decreasing trends from one trial to the next. 

III. F0 Generation Results 

 Log rank p-values, which were well above .05, indicated that there were no 

statistically significant differences between any doses within each trial, comparing 

time to first copulation. Time to first copulation decreased from one trial to the 

next. Dose 60, however, had the longest estimated copulation time for all three 

trials, but was never significantly greater than any of the other dose groups. 
30 

 

Dose

0 154.7 12.7 20% 121.6 16.5 42% 115.8 16.6 45%

30 144.0 12.5 30% 107.9 16.4 50% 110.5 17.6 45%

60 170.6 7.3 10% 159.8 11.2 15% 151.8 13.0 20%

125 150.3 13.4 20% 120.3 17.4 47% 122.1 17.3 40%

250 139.9 15.7 26% 124.3 17.1 35% 92.9 17.7 55%

Copulated

pairs

F0Generation ‐ Time to first Copulation

Log rank p‐value = .624 Log rank p‐value = .220 Log rank p‐value = .248

Trial 1 Trial 2 Trial 3

Average time

to first

copulation

Standard

Error

Average time

to first

copulation

Standard

Error Copulated

pairs

Average time

to first

copulation

Standard

Error Copulated

pairs
31 

IV. F1 Generation Results 

 For the first two trials, minimal, non-significant differences occurred between 

the doses. However, in trial 3, dose 30 had a significantly shorter copulation time 

estimate compared to dose 125. Dose 30 tended to have one of the shortest times 

in all three trials. 

V. F2 Generation Results 

 Significant differences between dose groups for copulation time occurred in 

trial 1 and trial 3. For both trials, the control group had a significantly longer 

copulation time than dose 30. The control group had one of the longest, and dose 

30 had the shortest copulation times for all three trials. Additionally, the only 

doses to shorten their copulation time through each trial were the control group 

and dose 250. 

 

Dose

0 142.7 13.5 30% 114.4 18.0 40% 91.2 17.8 60%

30 105.6 16.7 55% 113.4 16.0 50% 58.9 14.4 85%

60 110.2 14.7 55% 128.4 15.8 40% 100.5 16.9 60%

125 143.8 14.3 30% 117.6 17.2 40% 130.8 16.1 35%

250 135.3 15.2 35% 91.9 20.2 53% 123.3 18.2 39%

Average time

to first

copulation

Standard

Error

Copulated

pairs

Average time

to first

copulation

Standard

Error

Copulated

pairs

Average time

to first

copulation

Standard

Error

Copulated

pairs

F1 Generation ‐ Time to first Copulation

Log rank p‐value = .212 Log rank p‐value = .755 Log rank p‐value = .013

Trial 1 Trial 2 Trial 3

Dose

0 150.3 13.4 20% 112.8 17.2 50% 104.5 17.6 50%

30 72.8 14.4 80% 83.6 17.7 60% 44.5 13.6 85%

60 114.1 16.8 50% 111.8 17.7 45% 113.1 17.0 45%

125 118.2 16.6 45% 96.7 17.3 55% 96.7 16.0 60%

250 113.3 15.2 50% 105.7 16.1 60% 77.9 15.3 80%

Average time

to first

copulation

Standard

Error

Copulated

pairs

Average time

to first

copulation

Standard

Error

Copulated

pairs

Average time

to first

copulation

Standard

Error

Copulated

pairs

F2 Generation ‐ Time to first Copulation

Log rank p‐value = .005 Log rank p‐value = .863 Log rank p‐value = .005

Trial 1 Trial 2 Trial 3
32 

ENDOSULFAN EFFECTS ON ORGAN 

WEIGHTS

Vital Organ and Total Body Weights 

 This report analyzes the body and organ weights of Japanese quail that were 

collected and weighed after necropsy. Weights were taken for quails in one of the 

five dose groups ((control) 0 ppm, 30 ppm, 60 ppm, 125 ppm, 250 ppm) in all 

three generations (F0, F1, F2). Total body weight was measured as well as brain, 

kidney, liver, thyroid and coacal gland (CG) weights. CG weights were only 

taken for males in the F1 and F2 generations. SPSS version 16.0 was used to 

analyze the body and organ weight data. Statistical significance for this analysis 

was determined using an alpha (α) of .05 with a 95% confidence level. 

 
33 

I. Analysis Methods 

 An Analysis of Variance (ANOVA) test was used to determine if dose group 

averages for each measured organ weight were significantly different. An Fstatistic and corresponding p-value were calculated for each organ. If the p-value 

was below .05, then it was concluded that at least two dose groups had 

significantly different average weights for that organ. To find which dose groups 

had significantly different average weights, a Tukey post-hoc test was used. A 

Levene’s test was used to check to make sure the dose groups had equal 

variances, a major assumption that must be met in order to validly use ANOVA 

test results. If this assumption was violated (p-value for Levene’s test <.05), a 

non-parametric Kruskal-Wallis test was run instead of an ANOVA. 

II. Summary of Results 

 Statistically significant differences sporadically occurred throughout all three 

generations and in both genders. Testing at all three generations and both 

genders, thirty-two ANOVA tests were run to see if the organ weight averages 

were significantly different between the five dose groups, and statistically 

significant differences (p-value <.05) only occurred in six of the ANOVA tests. 

Additionally, for both males and females, rankings of dose groups based on their 

organ weights were very inconsistent across the three generations. Across all 

three generations, kidneys and liver weights were about twice as heavy for 

females as they were for males, and body weights were slightly higher for females 

also. Lastly, for each gender, organ weights from one generation to the next were 

similar and had no distinct differences. 

III. F0 Generation Results 

 A. Females 

No statistically significant differences (ANOVA p-values all above .05) in 

body or organ weights occurred between the five dose groups. Inconclusive 

trends in organ and body weight averages comparing the dosing level of 

endosulfan reaffirm that organ and body weights for female quails were not 

different in the F0 generation. Although no trend in average weights was present 

for the degree of endosulfan concentration, the control group had the lowest 

average for all organ and body weights except kidneys. 
34 

B. Males 

 Statistically significant differences in kidneys (p-value = .012) and thyroid (pvalue = .004) weights occurred between dose groups. For kidneys, the control 

group and dose 30 had significantly greater kidney weights compared to dose 250. 

For thyroid, the control group had a significantly greater thyroid weight compared 

to dose 125 and dose 250. Liver weights (p-value =.057) was also very close to 

having statistically significant differences between dose groups, with the greatest 

difference in dose groups being between dose 30 and dose 125. In general, it was 

evident that the control group and dose 30 usually had the two highest averages 

for the five organ weights. Additionally, dose 125 typically had the lowest 

average organ weight. 

 

C. Male and Female differences 

 Kidney and liver weight averages for males were about half of what they were 

for females. Body weight averages for males were also lower than females. 

Dose Average Std. Error Average Std. Error Average Std. Error Average Std. Error Average Std. Error

0 127.6 7.18 0.73 0.030 1.31 0.110 3.42 0.221 0.014 0.002

30 144.8 2.24 0.76 0.018 1.37 0.069 4.21 0.179 0.014 0.002

60 136.4 5.40 0.75 0.006 1.23 0.093 3.99 0.272 0.016 0.004

125 135.4 2.59 0.78 0.015 1.24 0.105 3.44 0.313 0.017 0.003

250 141.9 3.47 0.77 0.010 1.21 0.041 3.78 0.161 0.016 0.002

F0Generation Female Organ Weights

Body Weight Brain Kidneys Liver Thyroid

P‐value = .083* P‐value = .344 P‐value = .631 P‐value = .082 P‐value = .831

* P‐value calculated from Kruskall‐wallis test because homogenei ty of variance assumption violated in parametric ANOVA test.

Dose Average Std. Error Average Std. Error Average Std. Error Average Std. Error Average Std. Error

0 112.3 2.35 0.78 0.015 0.79 0.044 1.73 0.070 0.020 0.003

30 114.2 3.16 0.77 0.018 0.79 0.036 1.91 0.089 0.013 0.001

60† ‐ ‐ ‐ ‐ ‐ ‐ ‐ ‐ ‐ ‐

125 108.3 1.57 0.75 0.014 0.61 0.058 1.67 0.047 0.011 0.001

250 110.3 1.57 0.79 0.019 0.57 0.057 1.80 0.035 0.012 0.001

** Tukey post‐hoc results indicate that dose 0 has a significantly greater thyroid weight than dose 125 and dose

250.

† Only 1 observaƟon for dose 60.  With only 1 observaƟon, standard deviaƟon/error cannot be calculated and

therefore cannot be included in ANOVA test.

P‐value = .226 P‐value = .435 P‐value = .012* P‐value = .057 P‐value = .004**

* Tukey post‐hoc results indicate that dose 0 and dose 30 have significantly greater kidney weight than dose 250 at the 95%

confidence level.

F0Generation Male Organ Weights

Body Weight Brain Kidneys Liver Thyroid
35 

More variation in organ weight averages between dose groups occurred in the 

males compared to females, indicated by the lower p-values and greater variation 

in the averages across dose groups for many of the organ weights, excluding body 

weight. 

IV. F1 Generation 

A. Females 

 Statistically significant differences in body weight (p-value = .002) and brain 

weight (p-value = .038) occurred between dose groups. Dose 30, 125, and 250 

had significantly greater body weights compared to dose 60. For brain weights, 

dose 125 had a significantly greater average weight compared the control group. 

Overall, dose 60 had organ weights with the lowest averages. Conversely, dose 

125 had averages that were typically the highest for organ weights, especially for 

body weight, brain, and liver. 

B. Males 

 Statistically significant differences in thyroid (p-value = .033) and potentially, 

cloacal gland (p-value = .057) averages occurred between dose groups. However, 

post-hoc tests reveal that differences between dose groups were non-significant, 

due to the adjustment of multiple testing. Body weight, brain, kidney, and liver 

all have dose group averages that are minimally different with p-values well 

above .05. Overall, the control group’s averages for organ weights were typically 

the lowest average, excluding CG measurements. 

Dose Average Std. Error Average Std. Error Average Std. Error Average Std. Error Average Std. Error

0 137.0 4.24 0.73 0.009 1.39 0.003 4.10 0.206 0.014 0.004

30 142.2 2.09 0.76 0.013 1.28 0.072 4.10 0.243 0.011 0.001

60 129.7 2.64 0.75 0.006 1.30 0.058 3.69 0.226 0.008 0.001

125 144.7 1.32 0.78 0.010 1.32 0.027 4.16 0.241 0.012 0.002

250 144.4 3.21 0.75 0.030 1.30 0.026 4.10 0.281 0.016 0.003

† Kruskall‐wallis test indicates that dose 125 has a significantly greater brain weight averages compared to the control group

at 95% confidence level

F1Generation Female Organ Weights

Body Weight Brain Kidneys Liver Thyroid

P‐value = .002° P‐value = .038*† P‐value = .313* P‐value = .641 P‐value = .103*

* P‐value calculated from Kruskall‐wallis test because homogeneity of variance assumption violated in parametric ANOVA test.

° Tukey post‐hoc results indicate that dose 30, 250 and 125 have signi ficantly greater body weight average compared to dose

60 at 95% confidence level.
36 

C. Male and Female Differences 

 Kidney and liver weight averages for males were about half of what they were 

for females. Body weight averages for males were also lower than females. For 

males, dose 125’s averages are typically the highest for organ weights. 

V. F2 Generation 

A. Females 

 Minimal, non-significant differences in organ weight averages (p-values well 

above .05) occurred between dose groups. Additionally, no dose group(s) had 

consistently higher or lower averages compared to the other dose groups across all 

five organ weights. 

B. Males 

 Statistically significant differences in body weight occurred between dose 

groups. Post-hoc results indicated that dose 60 and dose 125 had significantly 

greater body weight averages compared to the control group. Besides thyroid, 

dose 125 typically had the highest average weight for all organs while dose 30 

typically had the lowest average weight. 

Dose Average Std. Error Average Std. Error Average Std. Error Average Std. Error Average Std. Error Average Std. Error

0 109.0 3.12 0.75 0.020 0.73 0.039 1.76 0.125 0.008 0.002 210.4 18.66

30 113.4 1.92 0.72 0.036 0.76 0.030 1.94 0.113 0.017 0.003 179.4 13.50

60 114.6 2.88 0.76 0.016 0.77 0.030 1.88 0.074 0.010 0.002 191.7 10.10

125 113.1 2.40 0.78 0.023 0.76 0.027 1.86 0.103 0.015 0.003 196.6 14.78

250 112.0 2.35 0.76 0.014 0.76 0.019 1.83 0.071 0.008 0.001 239.1 9.92

P‐value = .608 P‐value = .572 P‐value = .912 P‐value = .767 P‐value = .033 P‐value = .054

F1 Generation Male Organ Weights

Body Weight Brain Kidneys Liver Thyroid Coacal Gland

Dose Average Std. Error Average Std. Error Average Std. Error Average Std. Error Average Std. Error

0 138.1 2.72 0.77 0.009 1.19 0.035 4.02 0.192 0.014 0.0018

30 139.1 2.64 0.75 0.010 1.27 0.049 4.14 0.066 0.014 0.0010

60 132.8 1.53 0.75 0.013 1.33 0.035 4.43 0.367 0.011 0.0011

125 138.3 2.87 0.77 0.014 1.28 0.062 4.25 0.228 0.012 0.0019

250 135.8 1.81 0.77 0.011 1.22 0.053 3.98 0.377 0.015 0.0017

F2Generation Female Organ Weights

Body Weight Brain Kidneys Liver Thyroid

P‐value = .403* P‐value = .248 P‐value = .303 P‐value = .880* P‐value = .597

* P‐value calculated from Kruskall‐wallis test because homogeneity of variance assumption violated in parametric ANOVA test.
37 

C. Male and Female Differences 

 Just like in the F0 and F1 generations, kidney and liver weights for males were 

about half of what they were for females, and body weight averages for males 

were also lower than females. 

Dose Average Std. Error Average Std. Error Average Std. Error Average Std. Error Average Std. Error Average Std. Error

0 108.1 1.49 0.77 0.011 0.72 0.015 1.66 0.055 0.012 0.0080 221.1 14.23

30 111.1 2.41 0.76 0.015 0.77 0.018 1.79 0.053 0.011 0.0083 196.1 13.67

60 116.7 1.94 0.78 0.012 0.80 0.030 1.85 0.067 0.012 0.0086 201.3 6.43

125 117.4 1.79 0.79 0.017 0.79 0.025 1.92 0.072 0.010 0.0064 222.6 7.22

250 110.9 2.39 0.76 0.012 0.77 0.042 1.88 0.109 0.011 0.0074 201.4 16.48

* Tukey post‐hoc test indicate that dose 60 and dose 125 have signi ficantly greater body weight compared to dose 0 at a 95% confidence level.

P‐value = .009* P‐value = .322 P‐value = .345 P‐value = .157 P‐value = .894 P‐value = .432

F2Generation Male Organ Weights

Body Weight Brain Kidneys Liver Thyroid Coacal Gland
 

APPENDIX H 

EXPERIMENTAL PATHOLOGY LABORATORIES, INC., 

DRAFT PATHOLOGY REPORT 
 

BATTELLE PACIFIC-NORTHWEST DIVISION 

EPA CONTRACT NUMBER EP-W-06-025 (TO17, TD5) 

[PROTOCOL # 08JG-80-08-04-02] 

EPL PROJECT NUMBER 237-043 

PREPARE GLASS SLIDES AND HISTOLOGICALLY 

EVALUATE TISSUES FROM HATCHLING/SUBHATCHLING COTURNIX JAPONICA EXPOSED TO 

DIFFERENT CONCENTRATIONS OF ENDOSULFAN 

DRAFT PATHOLOGY REPORT 

Submitted by: 

Experimental Pathology Laboratories, Inc. 

P.O. Box 169 

Sterling, VA 20167-0169 

(703) 471-7060 

Submitted to: 

Battelle Pacific-Northwest Division 

Sequim, WA 98382 

March 3, 2010 
2 

TABLE OF CONTENTS 

Page 

PATHOLOGY SUMMARY .................................................................................1 

COMPLIANCE STATEMENT ............................................................................7 

QUALITY ASSURANCE FINAL CERTIFICATION ............................................8 

APPENDIX A: Figures .......................................................................................A-1 

APPENDIX B: Data Spreadsheet .....................................................................B-1 


 Experimental Pathology Laboratories, Inc. 

DRAFT PATHOLOGY SUMMARY 


 Experimental Pathology Laboratories, Inc. 

BATTELLE PACIFIC-NORTHWEST DIVISION 

EPA CONTRACT NUMBER EP-W-06-025 (TO17, TD5) 

[PROTOCOL # 08JG-80-08-04-02] 

EPL PROJECT NUMBER 237-043 

PREPARE GLASS SLIDES AND HISTOLOGICALLY EVALUATE 

TISSUES FROM HATCHLING/SUB-HATCHLING COTURNIX 

JAPONICA EXPOSED TO DIFFERENT CONCENTRATIONS OF 

ENDOSULFAN 

DRAFT PATHOLOGY REPORT 

INTRODUCTION

As part of the development and validation of a Tier 2, 2-generation avian toxicity 

test to characterize the adverse consequences of exposure to putative endocrine active 

substances, a histopathological evaluation was performed on tissues from hatchling/ subhatchling Japanese quail (Coturnix japonica) that were exposed to various concentrations 

of endosulfan via the diet (Table 1). 

Table 1. Number of Japanese Quail Hatchlings/Sub-hatchlings Evaluated 

Dose 

(ppm) No. of Males 

No. of 

Females 

Total 

Number 

0.0 8 4 12 

30 16 12 28 

60 13 16 29 

125 10 4 14 

250 9 9 18 

TOTAL 56 45 101 

METHODS

Necropsies were performed at the U.S. Army Center for Health Promotion & 

Preventive Medicine Directorate of Toxicology Health Effects Research. 

Tissues that were processed and evaluated included adrenal glands, brain within the 

skull (for medial preoptic nucleus [POM], pituitary, and pineal gland), Bursa of 

Fabricius, cloacal gland, kidney, liver, Mesonephric/Wolffian ducts (ductus deferens), 

Müllerian ducts (infundibulum, magnum, isthmus, shell gland, vagina), and testis. At 

EPL, tissues that contained bone (the skull, and samples of kidney, adrenal, and genital 

tissues that contained spine) were decalcified in a commercial formic acid/EDTA 

solution prior to routine processing for paraffin embedding. All processed and embedded 
- 2 - 



 Experimental Pathology Laboratories, Inc. 

Battelle Pacific-Northwest Division, 

EPA Contract Number EP-W-06-025 (TO17, TD5) 

[Protocol # 08JG-80-08-04-02] 

3/3/10 DRAFT 

tissues were microtomed to 4-6 m thick sections, and all sections were mounted on glass 

slides, stained with hematoxylin and eosin (H&E), and coverslipped according to routine 

methods. Sections were evaluated for treatment-induced effects via brightfield 

microscopy, and the prevalence and severity of findings were assessed as compared to 

controls. The severity of lesions was recorded as: Grade 1 = minimal, Grade 2 = 

slight/mild, Grade 3 = moderate, and Grade 4 = severe. Results were tabulated in an 

Excel spreadsheet (Appendix B). 

RESULTS

Based on incidence and severity data, there were no clear treatment-related findings 

in this study. 

There were a few findings that were only observed in endosulfan-treated quail, and 

these included: focally extensive hepatocellular necrosis (minimal to slight/mild) in one 

female of the 30 ppm dose group (Animal No. E070), two females of the 60 ppm dose 

group (Animal Nos. E080 and E082), and one female of the 125 ppm dose group (Animal 

No. E040); bile duct hyperplasia (minimal) and peribiliary fibrosis (minimal) in a female 

of the 250 ppm dose group (Animal No. E020); and a retained right ovary in a female of 

the 250 ppm dose group (Animal No. E061). 

In each instance, focally extensive hepatocellular necrosis (Figs. 1-4) was 

characterized by a discrete, roughly elliptical area of liver cell fragmentation, with 

varying degrees of secondary (dystrophic) mineralization of necrotic cellular debris. 

These lesions tended to be located proximate to, but not at, the liver surface, and were 

generally situated near the margins of liver lobes. 

Bile duct hyperplasia (Figs. 7, with Figs. 5 and 6 included for comparison) 

presented as increased bile duct profiles within portal triads, and peribiliary fibrosis was 

characterized by increased collagen in these same regions. 

The retained right ovary (Animal No. E061), which was smaller than the left, is 

illustrated in Fig. 8. 

Two other noteworthy or unusual findings that were clearly not treatment-related 

included a persistent Müllerian duct (Figs. 9-12) in a control male (Animal No. E065), 

and regionally extensive encephalomalacia in a control female (Animal No. E003) (Figs. 

14, 15, 17). 

It is possible that the single control male (Animal No. E065) that had a persistent 

Müllerian duct (Figs. 9-12) may actually have been female, as no gonadal tissue was 

recovered for that chick. The Müllerian duct is an embryonic precursor of the oviduct in 

females, whereas the paired Mesonephric/Wolffian ducts are forerunners of the ductus 

deferens. Both types of ducts are typically present in females at this stage of reproductive 
- 3 - 



 Experimental Pathology Laboratories, Inc. 

Battelle Pacific-Northwest Division, 

EPA Contract Number EP-W-06-025 (TO17, TD5) 

[Protocol # 08JG-80-08-04-02] 

3/3/10 DRAFT 

tract development (the Mesonephric/Wolffian ducts will subsequently atrophy); however, 

males are expected to have only the Mesonephric/Wolffian ducts. 

Regionally extensive encephalomalacia (brain necrosis) in the single control female 

(Animal No. E003) was characterized by well-defined, bilaterally symmetrical areas of 

neuropil vacuolation and rarefaction (Figs. 14, 15, 17). This loss of neuropil density was 

accompanied by necrosis of neuronal and glial cellular constituents, as evidenced by 

marked cytoplasmic shrinkage and dissolution accompanied by nuclear pyknosis. The 

prominence of small blood vessels in these regions was most likely a consequence of 

increased visual contrast as a function of decreased neuropil density. In the examined 

sections, areas of encephalomalacia extended throughout the dorsal cerebral cortex and 

were located within the granular layer and white matter of the cerebellar folia. Cerebellar 

folia were markedly collapsed as a result of the necrosis. 

Upon histologic examination of the gonads, two quail that were originally submitted 

as females (Animal Nos. E066 and E077) were determined to be males, and one animal 

originally submitted as male (Animal No. E079) was determined to be female. 

Other findings in this study generally occurred in comparable numbers of treated 

and control animals, or were low-frequency (incidental) findings that were observed in 

individual animals, and therefore not considered to be treatment-related. 

DISCUSSION AND CONCLUSIONS

In the present study, a cause of the focally-extensive hepatocellular necrosis in a 

low percentage of endosulfan-treated female quail was not apparent in the histologic 

sections. This type of change would not be expected to occur commonly as a 

spontaneous finding in Japanese quail embryos or hatchlings; however, the focal (vs. 

zonal) distribution of the necrosis would be more consistent with a nontoxic etiology 

such as infectious disease. It has been demonstrated that endosulfan administration can 

cause histopathologically evident liver lesions in fish and rodents (Altinok and Capkin, 

2007; Ballesteros et al., 2007; Glover et al., 2007; Singh et al., 2008). Commonly 

reported liver effects include degenerative vacuolar changes in hepatocytes and hepatic 

lipidosis (Ballesteros et al., 2007; Glover et al., 2007; Singh et al., 2008). In a prior 

report (EP-W-06-025; TO17, TD4), endosulfan exposure was clearly associated with 

macro- and microvesicular vacuolar degeneration of liver cells in adult female quail. 

However, the spatial distribution, extent and nature of the lesions in the present study 

differed markedly from the changes that were observed in adults (i.e., focal 

hepatocellular necrosis vs. zonal vacuolation), although it is interesting to note that in 

both age groups, these unusual types of liver changes were limited to females. 
- 4 - 



 Experimental Pathology Laboratories, Inc. 

Battelle Pacific-Northwest Division, 

EPA Contract Number EP-W-06-025 (TO17, TD5) 

[Protocol # 08JG-80-08-04-02] 

3/3/10 DRAFT 

The right ovary was retained in a female (Animal No. E061) of the 250 ppm dose 

group. Because this finding only occurred in a single animal, and because vestigial right 

ovaries have been observed in untreated Japanese quail (Fitzgerald, 1969), this result 

could not be associated definitively with endosulfan administration. Similarly, there is 

insufficient evidence that endosulfan treatment was responsible for the bile duct 

hyperplasia and peribiliary fibrosis in the single 250 ppm dose group female (Animal No. 

E020). 

Persistence of the right oviduct (Müllerian duct) is a documented effect of estrogen 

exposure in male Japanese quail (Berg et al., 1999; Ro and Kondo, 1979), although it has 

also been known to occur spontaneously in poultry (Wakamatsu et al., 2000). In the 

current study, the validity of this finding in one control male is uncertain, because 

gonadal tissue was not recovered for this animal, and thus the gender could not be 

confirmed. 

There was no histopathologically evident cause of the encephalomalacia in the 

single control female. A typical cause of this type of lesion in young poultry is vitamin E 

deficiency relative to the level of oxidized fatty acids in the feed (Riddell, 1987; Mézes et 

al., 1997). The common name for this condition is “crazy chick disease”. Although a 

nutritional etiology would seem unlikely for a sole animal in a laboratory study in which 

the diet did not vary, it is possible that this particular bird had an idiosyncratic response 

related to some anomaly of vitamin E absorption or fatty acid metabolism. 

JEFFREY C. WOLF, DVM, Diplomate, ACVP Date 

Veterinary Pathologist 

JCW/cb 

 
- 5 - 



 Experimental Pathology Laboratories, Inc. 

Battelle Pacific-Northwest Division, 

EPA Contract Number EP-W-06-025 (TO17, TD5) 

[Protocol # 08JG-80-08-04-02] 

3/3/10 DRAFT 

REFERENCES

Altinok I, Capkin E (2007) Histopathology of rainbow trout exposed to sublethal 

concentrations of methiocarb or endosulfan. Toxicol Pathol, 35(3):405-410. 

Ballesteros ML, Bianchi GE, Carranza M, Bistoni MA (2007) Endosulfan acute toxicity 

and histomorphological alterations in Jenynsia multidentata (Anablepidae, 

Cyprinodontiformes). J Environ Sci Health B, 42(4):351-357. 

Berg C, Halldin K, Fridolfsson A-K, Brandt I, Brunström B (1999) The avian egg as a 

test system for endocrine disrupters: effects of diethylstilbestrol and ethynylestradiol on 

sex organ development. Sci Total Environ, 233(1-3):57-66. 

Fitzgerald TC (1969) The Coturnix Quail: Anatomy and Histology. The Iowa State 

University Press, Ames, Iowa, p. 260. 

Glover CN, Petri D, Tollefsen K-E, Jorum N, Handy RD, Berntssen MHG (2007) 

Assessing the sensitivity of Atlantic salmon (Salmo salar) to dietary endosulfan exposure 

using tissue biochemistry and histology. Aquat Toxicol, 84(3):346-355. 

Mézes M, Surai P, Sályi G, Speake BK, Gaál T, Maldjian A (1997) Nutritional metabolic 

diseases of poultry and disorders of the biological antioxidant defense system. Acta Vet 

Hung, 45(3):349-360. 

Riddell C (1987) Avian Histopathology. American Association of Avian Pathologists, 

Inc., Allen Press, Inc., Lawrence, KS, p. 76. 

Ro S, Kondo K (1979) The effects of prenatal treatment of exogenous estrogen on the 

postnatal development of the sexual organs and reproductive ability in the male Japanese 

quail. Jap J Zootech Sci, 50(11):821-832. 

Singh ND, Sharma AK, Dwivedi P, Patil RD, Kumar M (2008) Experimentally induced 

citrinin and endosulfan toxicity in pregnant Wistar rats: histopathological alterations in 

liver and kidneys of fetuses. J Appl Toxicol, 28(7):901-907. 

Wakamatsu M, Yamagata T, Namikawa T, Mizutani M (2000) Hereditary persistent right 

oviduct in the chicken PNP/DO line. Poult Sci, 79(8):1075-1081. 
- 6 - 



 Experimental Pathology Laboratories, Inc. 

3/3/10 DRAFT 

COMPLIANCE STATEMENT 

Client 

Name 

Battelle Pacific-Northwest 

Division 

EPL Principal 

Investigator Dr. Jeffrey C. Wolf 

Client 

Study 

EPA EP-W-06-025 (TO17, TD5) 

[Protocol # 08JG-80-08-04-02] 

EPL 

Pathologist Dr. Jeffrey C. Wolf 

Species 

Coturnix japonica

(Japanese quail) 

EPL Project 

Number 237-043 

Study 

Title 

Prepare Glass Slides and Histologically Evaluate Tissues From Hatchling/Subhatchling Coturnix japonica Exposed to Different Concentrations of Endosulfan 

 

Test 

Article Endosulfan 

The Histopathology portions of the above-referenced study were conducted in 

compliance with the Good Laboratory Practice regulations of the Environmental 

Protection Agency as stipulated by 40 CFR 160. 

 EPL Principal Investigator 

 

 Date 
- 7 - 



 Experimental Pathology Laboratories, Inc. 

3/3/10 DRAFT 

QUALITY ASSURANCE FINAL CERTIFICATION 

Study Title: Prepare Glass Slides and Histologically Evaluate Tissues From Hatchling/Subhatchling Coturnix japonica Exposed to Different Concentrations of Endosulfan 

Client Study: EPA EP-W-06-025 (TO17, TD5) EPL Principal Investigator: Dr. Jeffrey C. Wolf 

[Protocol # 08JG-80-08-04-02] 

EPL Project Number: 237-043 EPL Pathologist: Dr. Jeffrey C. Wolf 

The following aspects of this study were inspected by the Quality Assurance Unit of Experimental 

Pathology Laboratories, Inc. Dates inspections were performed and findings reported to the EPL Principal 

Investigator and Management are indicated below. 

 Dates 

Area Inspected Inspection Reporting 

EPL Project Sheets 

Project Setup 

In-Process 

Data Review 

Draft Pathology Report 

Final Pathology Report 

Date of last quarterly facility inspection 11/09 

 

EPL Quality Assurance Unit Date 
APPENDIX A 

FIGURES 
A-1 

FIGURES

Figure 1 (Z15301). Liver from a female quail treated with 60 ppm endosulfan (Animal 

No. E080). There is a large, well-circumscribed area of necrosis near the liver margin 

(arrows). H&E. 
A-2 

Figure 2 (Z15302). Higher magnification of liver from preceding figure. Affected 

hepatocytes are fragmented and dissociated. The lack of any appreciable inflammatory 

response suggests that this lesion is of short term duration. H&E. 
A-3 

Figure 3 (Z15303). Liver from a female treated with 125 ppm endosulfan (Animal No. 

E040). Another example of focal liver necrosis. H&E. 
A-4 

Figure 4 (Z15304). Higher magnification of liver from preceding figure. 

Mineralization (dense dark purple deposits; arrows) of necrotic tissue is more obvious in 

this animal’s liver compared to the previous example. H&E. 
A-5 

Figure 5 (Z15299). Liver from a control male (Animal No. E066). At this age, portal 

triads often contain multiple bile duct profiles (arrows) that are surrounded by a matrix 

of loose fibrous connective tissue. H&E. 
A-6 

Figure 6 (Z15300). Higher magnification of liver from preceding figure to illustrate 

bile duct profile (B) and fibrous connective tissue (F). H&E. 

B 

F 
A-7 

Figure 7 (Z15322). Liver from a female treated with 250 ppm endosulfan (Animal No. 

E020). Bile duct profiles (B) are more numerous and tortuous than was generally 

observed in controls, and portal triads are expanded by more abundant fibrosis (F). 

These particular changes were only observed in a single high dose female. H&E. 

B 

B 

B 

B 

F 

F 
A-8 

Figure 8 (Z15321). Transverse section through the anterior abdomen of a female 

treated with 250 ppm endosulfan (Animal No. E061). Evident are spine (S), kidneys 

(K), adrenals (A), and ovaries (LO = left ovary, RO = right ovary). Retained right ovary 

can be an indication of exposure to compounds with estrogenic activity, but in this case 

relationship to treatment is questionable, as only a single animal was affected, and there 

is apparently some degree of spontaneous right ovary retention in Japanese quail. H&E. 

K K 

A A 

RO 

LO 

S 
A-9 

Figure 9 (Z15317). Transverse section through the mid abdomen of a bird listed as a 

control male (Animal No. E065). A precursor to the oviduct called the Müllerian duct 

(arrow) is evident in its typical location anterior and lateral to the left metanephric 

kidney (K). Although this could be an endocrine disruptive-type finding, it is also 

possible that this animal’s gender was simply misidentified. H&E. 

K 
A-10 

Figure 10 (Z15318). Higher magnification of Müllerian duct from preceding figure. 

Characteristically, the wall of this duct is thick and densely cellular. H&E. 
A-11 

Figure 11 (Z15313). Another transverse section at a slightly different level through the 

mid abdomen of Animal No. E065. The Müllerian duct (closed arrow) is located lateral 

to the metanephric (urinary) duct (open arrow). H&E. 
A-12 

Figure 12 (Z15315). Yet another transverse section at a slightly different level through 

the mid abdomen of Animal No. E065. At this level the Müllerian duct is not evident, 

but the Mesonephric/Wolffian duct is present (closed arrow) medial to the metanephric 

duct (open arrow). This figure does not illustrate any abnormality, but is included for 

the purpose of anatomic comparison. The Mesonephric/Wolffian duct, which is the 

precursor of the ductus deferens in males, is also present in females at this age, but will 

subsequently atrophy. H&E. 
A-13 

Figure 13 (Z15305). Transverse section through the brain of a control female (Animal 

No. E018). CC = cerebral cortex, LV = lateral ventricle, TV = third ventricle, E = eye. 

H&E. 

E E

LV LV 

TV 

CC CC
A-14 

Figure 14 (Z15307). Transverse section through the brain of another control female 

(Animal No. E003). Regionally extensive encephalomalacia (brain necrosis) is evident 

as bilaterally symmetrical pale staining areas in the dorsal cerebral cortex. 

CC = cerebral cortex, LV = lateral ventricle, TV = third ventricle, E = eye. H&E. 

CC CC

LV 

LV 

TV 

E 

E 
A-15 

Figure 15 (Z15309). Higher magnification of brain from previous figure, at border 

between less affected (LA) and encephalomalacic (E) regions. H&E. 

E 

LA 
A-16 

 

Figure 16 (Z15306). Sagittal section through the brain of control female Animal No. 

E018 to demonstrate the normal cerebellar folia (CF). H&E. 

CF 

CF 

CF 
A-17 

Figure 17 (Z15310). Sagittal section through the brain of control female with 

encephalomalacia (Animal No. E003). Cerebellar folia (CF) are markedly collapsed, 

most of the granular layer has disappeared, and the density of the white matter is 

decreased. When it is a consequence of vitamin E deficiency, the common name for this 

disorder is “crazy chick disease”. H&E. 

CF 

CF 

CF 
APPENDIX B 

DATA SPREADSHEET 
B-1 

Group Tissue Lesion 

Severity 

Grade 

Images 

Taken 

0 ppm ADRENAL No Section . . 

0 ppm BURSA Apoptotic Necrosis, Increased 1 . 

0 ppm CLOACAL GLAND . . . 

0 ppm KIDNEY . . . 

0 ppm LIVER . . . 

0 ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

0 ppm MULLERIAN DUCTS-M Persistent P . 

0 ppm PINEAL GLAND No Section . . 

0 ppm PITUITARY . . . 

0 ppm PREOPTIC NUCLEUS . . . 

0 ppm TESTIS No Section . . 

0 ppm ADRENAL No Section . . 

0 ppm BURSA . . . 

0 ppm CLOACAL GLAND . . . 

0 ppm KIDNEY . . . 

0 ppm LIVER . . . 

0 ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

0 ppm PINEAL GLAND . . . 

0 ppm PITUITARY . . . 

0 ppm PREOPTIC NUCLEUS . . . 

0 ppm TESTIS No Section . . 

0 ppm ADRENAL No Section . . 

0 ppm BURSA . . . 

0 ppm CLOACAL GLAND . . . 

0 ppm KIDNEY . . . 

0 ppm LIVER . . . 

0 ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

0 ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS Missing one of pair . . 

0 ppm PINEAL GLAND . . . 

0 ppm PITUITARY Cyst(s) P . 

0 ppm PREOPTIC NUCLEUS . . . 

0 ppm TESTIS . . . 

0 ppm TESTIS Missing one of pair . . 

0 ppm ADRENAL . . . 

0 ppm BURSA . . . 

0 ppm CLOACAL GLAND . . . 

0 ppm KIDNEY . . . 

0 ppm LIVER . . . 

0 ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

0 ppm PINEAL GLAND . . . 

0 ppm PITUITARY Cyst(s) P . 

0 ppm PREOPTIC NUCLEUS . . . 

0 ppm TESTIS . . . 
B-1 

0 ppm ADRENAL . . . 

0 ppm BURSA No Section . . 

0 ppm CLOACAL GLAND . . . 

0 ppm KIDNEY Tubular Dilation 1 . 

0 ppm LIVER . . . 

0 ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

0 ppm PINEAL GLAND . . . 

0 ppm PITUITARY Cyst(s) P . 

0 ppm PREOPTIC NUCLEUS . . . 

0 ppm TESTIS . . . 

0 ppm ADRENAL . . . 

0 ppm BURSA . . . 

0 ppm CLOACAL GLAND . . . 

0 ppm KIDNEY . . . 

0 ppm LIVER . . . 

0 ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

0 ppm PINEAL GLAND No Section . . 

0 ppm PITUITARY . . . 

0 ppm PREOPTIC NUCLEUS . . . 

0 ppm TESTIS . . . 

0 ppm ADRENAL . . . 

0 ppm BURSA . . . 

0 ppm CLOACAL GLAND . . . 

0 ppm KIDNEY . . . 

0 ppm LIVER . . . 

0 ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . 1 

0 ppm MULLERIAN DUCTS-M Persistent P 3 

0 ppm PINEAL GLAND No Section . . 

0 ppm PITUITARY No Section . . 

0 ppm PREOPTIC NUCLEUS . . . 

0 ppm TESTIS No Section . . 

0 ppm ADRENAL No Section . . 

0 ppm BURSA . . . 

0 ppm CLOACAL GLAND . . . 

0 ppm KIDNEY . . . 

0 ppm LIVER . . 2 

0 ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

0 ppm PINEAL GLAND No Section . . 

0 ppm PITUITARY . . . 

0 ppm PREOPTIC NUCLEUS . . . 

0 ppm TESTIS Apoptotic Necrosis, Increased 1 . 

30 

ppm ADRENAL . . . 

30 

ppm BURSA . . . 

30 

ppm CLOACAL GLAND . . . 
B-1 

30 

ppm KIDNEY . . . 

30 

ppm LIVER . . . 

30 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

30 

ppm PINEAL GLAND . . . 

30 

ppm PITUITARY . . . 

30 

ppm PREOPTIC NUCLEUS . . . 

30 

ppm TESTIS . . . 

30 

ppm TESTIS Missing one of pair . . 

30 

ppm ADRENAL . . . 

30 

ppm ADRENAL Missing one of pair . . 

30 

ppm BURSA . . . 

30 

ppm CLOACAL GLAND . . . 

30 

ppm KIDNEY . . . 

30 

ppm LIVER . . . 

30 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

30 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS Missing one of pair . . 

30 

ppm PINEAL GLAND No Section . . 

30 

ppm PITUITARY . . . 

30 

ppm PREOPTIC NUCLEUS . . . 

30 

ppm TESTIS . . . 

30 

ppm TESTIS Missing one of pair . . 

30 

ppm ADRENAL . . . 

30 

ppm BURSA Apoptotic Necrosis, Increased 1 . 

30 

ppm CLOACAL GLAND . . . 

30 

ppm KIDNEY Tubular Dilation 1 . 

30 

ppm LIVER . . . 

30 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

30 PINEAL GLAND No Section . . 
B-1 

ppm 

30 

ppm PITUITARY . . . 

30 

ppm PREOPTIC NUCLEUS . . . 

30 

ppm TESTIS . . . 

30 

ppm TESTIS Missing one of pair . . 

30 

ppm ADRENAL . . . 

30 

ppm BURSA . . . 

30 

ppm CLOACAL GLAND . . . 

30 

ppm KIDNEY . . . 

30 

ppm LIVER . . . 

30 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

30 

ppm PINEAL GLAND No Section . . 

30 

ppm PITUITARY Cyst(s) P . 

30 

ppm PREOPTIC NUCLEUS . . . 

30 

ppm TESTIS . . . 

30 

ppm ADRENAL . . . 

30 

ppm BURSA . . . 

30 

ppm CLOACAL GLAND . . . 

30 

ppm KIDNEY . . . 

30 

ppm LIVER . . . 

30 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

30 

ppm PINEAL GLAND No Section . . 

30 

ppm PITUITARY . . . 

30 

ppm PREOPTIC NUCLEUS . . . 

30 

ppm TESTIS . . . 

30 

ppm TESTIS Missing one of pair . . 

30 

ppm ADRENAL . . . 

30 

ppm BURSA No Section . . 
B-1 

30 

ppm CLOACAL GLAND . . . 

30 

ppm KIDNEY . . . 

30 

ppm LIVER . . . 

30 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

30 

ppm PINEAL GLAND No Section . . 

30 

ppm PITUITARY Cyst(s) P . 

30 

ppm PREOPTIC NUCLEUS . . . 

30 

ppm TESTIS . . . 

30 

ppm ADRENAL . . . 

30 

ppm BURSA Apoptotic Necrosis, Increased 1 . 

30 

ppm CLOACAL GLAND . . . 

30 

ppm KIDNEY . . . 

30 

ppm LIVER . . . 

30 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

30 

ppm PINEAL GLAND . . . 

30 

ppm PITUITARY . . . 

30 

ppm PREOPTIC NUCLEUS . . . 

30 

ppm TESTIS . . . 

30 

ppm TESTIS Missing one of pair . . 

30 

ppm ADRENAL . . . 

30 

ppm BURSA . . . 

30 

ppm CLOACAL GLAND . . . 

30 

ppm KIDNEY . . . 

30 

ppm LIVER . . . 

30 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

30 

ppm PINEAL GLAND No Section . . 

30 

ppm PITUITARY . . . 

30 PREOPTIC NUCLEUS . . . 
B-1 

ppm 

30 

ppm TESTIS . . . 

30 

ppm ADRENAL . . . 

30 

ppm BURSA . . . 

30 

ppm CLOACAL GLAND . . . 

30 

ppm KIDNEY . . . 

30 

ppm LIVER . . . 

30 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

30 

ppm PINEAL GLAND No Section . . 

30 

ppm PITUITARY . . . 

30 

ppm PREOPTIC NUCLEUS . . . 

30 

ppm TESTIS No Section . . 

30 

ppm ADRENAL . . . 

30 

ppm BURSA Apoptotic Necrosis, Increased 1 . 

30 

ppm CLOACAL GLAND . . . 

30 

ppm KIDNEY . . . 

30 

ppm LIVER . . . 

30 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

30 

ppm PINEAL GLAND No Section . . 

30 

ppm PITUITARY Cyst(s) P . 

30 

ppm PREOPTIC NUCLEUS . . . 

30 

ppm TESTIS . . . 

30 

ppm ADRENAL . . . 

30 

ppm BURSA Apoptotic Necrosis, Increased 1 . 

30 

ppm CLOACAL GLAND . . . 

30 

ppm KIDNEY . . . 

30 

ppm LIVER . . . 

30 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 
B-1 

30 

ppm PINEAL GLAND No Section . . 

30 

ppm PITUITARY . . . 

30 

ppm PREOPTIC NUCLEUS . . . 

30 

ppm TESTIS . . . 

30 

ppm ADRENAL . . . 

30 

ppm BURSA . . . 

30 

ppm CLOACAL GLAND . . . 

30 

ppm KIDNEY . . . 

30 

ppm LIVER . . . 

30 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

30 

ppm PINEAL GLAND No Section . . 

30 

ppm PITUITARY . . . 

30 

ppm PREOPTIC NUCLEUS . . . 

30 

ppm TESTIS . . . 

30 

ppm ADRENAL . . . 

30 

ppm BURSA . . . 

30 

ppm CLOACAL GLAND . . . 

30 

ppm KIDNEY . . . 

30 

ppm LIVER . . . 

30 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

30 

ppm PINEAL GLAND No Section . . 

30 

ppm PITUITARY . . . 

30 

ppm PREOPTIC NUCLEUS . . . 

30 

ppm TESTIS . . . 

30 

ppm TESTIS Missing one of pair . . 

30 

ppm ADRENAL . . . 

30 

ppm BURSA . . . 

30 CLOACAL GLAND . . . 
B-1 

ppm 

30 

ppm KIDNEY . . . 

30 

ppm LIVER . . . 

30 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

30 

ppm PINEAL GLAND . . . 

30 

ppm PITUITARY . . . 

30 

ppm PREOPTIC NUCLEUS . . . 

30 

ppm TESTIS . . . 

30 

ppm ADRENAL . . . 

30 

ppm BURSA . . . 

30 

ppm CLOACAL GLAND . . . 

30 

ppm KIDNEY . . . 

30 

ppm LIVER . . . 

30 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

30 

ppm PINEAL GLAND . . . 

30 

ppm PITUITARY Cyst(s) P . 

30 

ppm PREOPTIC NUCLEUS . . . 

30 

ppm TESTIS Apoptotic Necrosis, Increased 1 . 

30 

ppm TESTIS Missing one of pair . . 

30 

ppm ADRENAL . . . 

30 

ppm BURSA . . . 

30 

ppm CLOACAL GLAND . . . 

30 

ppm KIDNEY . . . 

30 

ppm LIVER . . . 

30 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

30 

ppm PINEAL GLAND . . . 

30 

ppm PITUITARY . . . 

30 

ppm PREOPTIC NUCLEUS . . . 
B-1 

30 

ppm TESTIS . . . 

60 

ppm ADRENAL . . . 

60 

ppm BURSA . . . 

60 

ppm CLOACAL GLAND . . . 

60 

ppm KIDNEY . . . 

60 

ppm LIVER . . . 

60 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

60 

ppm PINEAL GLAND No Section . . 

60 

ppm PITUITARY . . . 

60 

ppm PREOPTIC NUCLEUS . . . 

60 

ppm TESTIS . . . 

60 

ppm TESTIS Missing one of pair . . 

60 

ppm ADRENAL . . . 

60 

ppm BURSA . . . 

60 

ppm CLOACAL GLAND . . . 

60 

ppm KIDNEY . . . 

60 

ppm LIVER . . . 

60 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

60 

ppm PINEAL GLAND No Section . . 

60 

ppm PITUITARY Cyst(s) P . 

60 

ppm PREOPTIC NUCLEUS . . . 

60 

ppm TESTIS . . . 

60 

ppm TESTIS Missing one of pair . . 

60 

ppm ADRENAL . . . 

60 

ppm BURSA . . . 

60 

ppm CLOACAL GLAND . . . 

60 

ppm KIDNEY . . . 

60 LIVER . . . 
B-1 

ppm 

60 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

60 

ppm PINEAL GLAND No Section . . 

60 

ppm PITUITARY . . . 

60 

ppm PREOPTIC NUCLEUS . . . 

60 

ppm TESTIS . . . 

60 

ppm TESTIS Missing one of pair . . 

60 

ppm ADRENAL . . . 

60 

ppm BURSA . . . 

60 

ppm CLOACAL GLAND . . . 

60 

ppm KIDNEY . . . 

60 

ppm LIVER . . . 

60 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

60 

ppm PINEAL GLAND . . . 

60 

ppm PITUITARY . . . 

60 

ppm PREOPTIC NUCLEUS . . . 

60 

ppm TESTIS . . . 

60 

ppm ADRENAL . . . 

60 

ppm BURSA . . . 

60 

ppm CLOACAL GLAND . . . 

60 

ppm KIDNEY . . . 

60 

ppm LIVER . . . 

60 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

60 

ppm PINEAL GLAND No Section . . 

60 

ppm PITUITARY . . . 

60 

ppm PREOPTIC NUCLEUS . . . 

60 

ppm TESTIS . . . 

60 

ppm ADRENAL . . . 
B-1 

60 

ppm BURSA . . . 

60 

ppm CLOACAL GLAND . . . 

60 

ppm KIDNEY . . . 

60 

ppm LIVER . . . 

60 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

60 

ppm PINEAL GLAND . . . 

60 

ppm PITUITARY . . . 

60 

ppm PREOPTIC NUCLEUS . . . 

60 

ppm TESTIS . . . 

60 

ppm ADRENAL . . . 

60 

ppm BURSA Apoptotic Necrosis, Increased 1 . 

60 

ppm CLOACAL GLAND . . . 

60 

ppm KIDNEY . . . 

60 

ppm LIVER . . . 

60 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

60 

ppm PINEAL GLAND . . . 

60 

ppm PITUITARY . . . 

60 

ppm PREOPTIC NUCLEUS . . . 

60 

ppm TESTIS . . . 

60 

ppm TESTIS Missing one of pair . . 

60 

ppm ADRENAL . . . 

60 

ppm BURSA . . . 

60 

ppm CLOACAL GLAND . . . 

60 

ppm KIDNEY . . . 

60 

ppm LIVER . . . 

60 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

60 

ppm PINEAL GLAND . . . 

60 PITUITARY . . . 
B-1 

ppm 

60 

ppm PREOPTIC NUCLEUS . . . 

60 

ppm TESTIS . . . 

60 

ppm TESTIS Missing one of pair . . 

60 

ppm ADRENAL . . . 

60 

ppm BURSA . . . 

60 

ppm CLOACAL GLAND . . . 

60 

ppm KIDNEY . . . 

60 

ppm LIVER . . . 

60 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

60 

ppm PINEAL GLAND No Section . . 

60 

ppm PITUITARY No Section . . 

60 

ppm PREOPTIC NUCLEUS . . . 

60 

ppm TESTIS . . . 

60 

ppm TESTIS Missing one of pair . . 

60 

ppm ADRENAL . . . 

60 

ppm BURSA . . . 

60 

ppm CLOACAL GLAND . . . 

60 

ppm KIDNEY . . . 

60 

ppm LIVER . . . 

60 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

60 

ppm PINEAL GLAND No Section . . 

60 

ppm PITUITARY No Section . . 

60 

ppm PREOPTIC NUCLEUS . . . 

60 

ppm TESTIS . . . 

60 

ppm ADRENAL . . . 

60 

ppm BURSA . . . 

60 

ppm CLOACAL GLAND . . . 
B-1 

60 

ppm KIDNEY . . . 

60 

ppm LIVER . . . 

60 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

60 

ppm PINEAL GLAND No Section . . 

60 

ppm PITUITARY . . . 

60 

ppm PREOPTIC NUCLEUS . . . 

60 

ppm TESTIS No Section . . 

60 

ppm ADRENAL . . . 

60 

ppm BURSA . . . 

60 

ppm CLOACAL GLAND . . . 

60 

ppm KIDNEY . . . 

60 

ppm LIVER . . . 

60 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

60 

ppm PINEAL GLAND No Section . . 

60 

ppm PITUITARY . . . 

60 

ppm PREOPTIC NUCLEUS . . . 

60 

ppm TESTIS . . . 

60 

ppm TESTIS Missing one of pair . . 

60 

ppm ADRENAL . . . 

60 

ppm BURSA . . . 

60 

ppm CLOACAL GLAND . . . 

60 

ppm KIDNEY . . . 

60 

ppm LIVER . . . 

60 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

60 

ppm PINEAL GLAND No Section . . 

60 

ppm PITUITARY Cyst(s) P . 

60 

ppm PREOPTIC NUCLEUS . . . 

60 TESTIS Apoptotic Necrosis, Increased 1 . 
B-1 

ppm 

60 

ppm TESTIS Missing one of pair . . 

125 

ppm ADRENAL . . . 

125 

ppm BURSA . . . 

125 

ppm CLOACAL GLAND . . . 

125 

ppm KIDNEY . . . 

125 

ppm LIVER . . . 

125 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

125 

ppm PINEAL GLAND No Section . . 

125 

ppm PITUITARY . . . 

125 

ppm PREOPTIC NUCLEUS . . . 

125 

ppm TESTIS . . . 

125 

ppm ADRENAL . . . 

125 

ppm BURSA . . . 

125 

ppm CLOACAL GLAND . . . 

125 

ppm KIDNEY . . . 

125 

ppm LIVER . . . 

125 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

125 

ppm PINEAL GLAND . . . 

125 

ppm PITUITARY . . . 

125 

ppm PREOPTIC NUCLEUS . . . 

125 

ppm TESTIS No Section . . 

125 

ppm ADRENAL . . . 

125 

ppm BURSA . . . 

125 

ppm CLOACAL GLAND . . . 

125 

ppm KIDNEY . . . 

125 

ppm LIVER . . . 

125 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 
B-1 

125 

ppm PINEAL GLAND . . . 

125 

ppm PITUITARY . . . 

125 

ppm PREOPTIC NUCLEUS . . . 

125 

ppm TESTIS . . . 

125 

ppm ADRENAL . . . 

125 

ppm BURSA No Section . . 

125 

ppm CLOACAL GLAND . . . 

125 

ppm KIDNEY . . . 

125 

ppm LIVER . . . 

125 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

125 

ppm PINEAL GLAND . . . 

125 

ppm PITUITARY . . . 

125 

ppm PREOPTIC NUCLEUS . . . 

125 

ppm TESTIS . . . 

125 

ppm TESTIS Missing one of pair . . 

125 

ppm ADRENAL . . . 

125 

ppm BURSA . . . 

125 

ppm CLOACAL GLAND . . . 

125 

ppm KIDNEY . . . 

125 

ppm LIVER . . . 

125 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

125 

ppm PINEAL GLAND No Section . . 

125 

ppm PITUITARY . . . 

125 

ppm PREOPTIC NUCLEUS . . . 

125 

ppm TESTIS . . . 

125 

ppm ADRENAL . . . 

125 

ppm BURSA . . . 

125 CLOACAL GLAND . . . 
B-1 

ppm 

125 

ppm KIDNEY . . . 

125 

ppm LIVER . . . 

125 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

125 

ppm PINEAL GLAND . . . 

125 

ppm PITUITARY . . . 

125 

ppm PREOPTIC NUCLEUS . . . 

125 

ppm TESTIS . . . 

125 

ppm ADRENAL . . . 

125 

ppm BURSA . . . 

125 

ppm CLOACAL GLAND . . . 

125 

ppm KIDNEY . . . 

125 

ppm LIVER . . . 

125 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

125 

ppm PINEAL GLAND . . . 

125 

ppm PITUITARY . . . 

125 

ppm PREOPTIC NUCLEUS . . . 

125 

ppm TESTIS . . . 

125 

ppm TESTIS Missing one of pair . . 

125 

ppm ADRENAL . . . 

125 

ppm BURSA . . . 

125 

ppm CLOACAL GLAND . . . 

125 

ppm KIDNEY . . . 

125 

ppm LIVER . . . 

125 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

125 

ppm PINEAL GLAND No Section . . 

125 

ppm PITUITARY . . . 

125 

ppm PREOPTIC NUCLEUS . . . 
B-1 

125 

ppm TESTIS . . . 

125 

ppm TESTIS Missing one of pair . . 

125 

ppm ADRENAL No Section . . 

125 

ppm BURSA . . . 

125 

ppm CLOACAL GLAND . . . 

125 

ppm KIDNEY . . . 

125 

ppm LIVER . . . 

125 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

125 

ppm PINEAL GLAND No Section . . 

125 

ppm PITUITARY . . . 

125 

ppm PREOPTIC NUCLEUS . . . 

125 

ppm TESTIS . . . 

125 

ppm TESTIS Missing one of pair . . 

125 

ppm ADRENAL . . . 

125 

ppm BURSA . . . 

125 

ppm CLOACAL GLAND . . . 

125 

ppm KIDNEY . . . 

125 

ppm LIVER . . . 

125 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

125 

ppm PINEAL GLAND No Section . . 

125 

ppm PITUITARY . . . 

125 

ppm PREOPTIC NUCLEUS . . . 

125 

ppm TESTIS No Section . . 

250 

ppm ADRENAL . . . 

250 

ppm BURSA . . . 

250 

ppm CLOACAL GLAND . . . 

250 

ppm KIDNEY . . . 

250 LIVER . . . 
B-1 

ppm 

250 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

250 

ppm PINEAL GLAND No Section . . 

250 

ppm PITUITARY . . . 

250 

ppm PREOPTIC NUCLEUS . . . 

250 

ppm TESTIS . . . 

250 

ppm TESTIS Missing one of pair . . 

250 

ppm ADRENAL . . . 

250 

ppm BURSA . . . 

250 

ppm CLOACAL GLAND . . . 

250 

ppm KIDNEY . . . 

250 

ppm LIVER 

Hepatocellular Vacuolation, Diffuse, Lipid 

Type 1 . 

250 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

250 

ppm PINEAL GLAND No Section . . 

250 

ppm PITUITARY Cyst(s) P . 

250 

ppm PREOPTIC NUCLEUS . . . 

250 

ppm TESTIS . . . 

250 

ppm ADRENAL . . . 

250 

ppm BURSA . . . 

250 

ppm CLOACAL GLAND . . . 

250 

ppm KIDNEY . . . 

250 

ppm LIVER . . . 

250 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

250 

ppm PINEAL GLAND . . . 

250 

ppm PITUITARY . . . 

250 

ppm PREOPTIC NUCLEUS . . . 

250 

ppm TESTIS . . . 

250 

ppm ADRENAL . . . 
B-1 

250 

ppm BURSA No Section . . 

250 

ppm CLOACAL GLAND . . . 

250 

ppm KIDNEY . . . 

250 

ppm LIVER . . . 

250 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

250 

ppm PINEAL GLAND No Section . . 

250 

ppm PITUITARY . . . 

250 

ppm PREOPTIC NUCLEUS . . . 

250 

ppm TESTIS . . . 

250 

ppm ADRENAL . . . 

250 

ppm BURSA . . . 

250 

ppm CLOACAL GLAND . . . 

250 

ppm KIDNEY . . . 

250 

ppm LIVER . . . 

250 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

250 

ppm PINEAL GLAND No Section . . 

250 

ppm PITUITARY . . . 

250 

ppm PREOPTIC NUCLEUS . . . 

250 

ppm TESTIS . . . 

250 

ppm ADRENAL . . . 

250 

ppm BURSA . . . 

250 

ppm CLOACAL GLAND . . . 

250 

ppm KIDNEY . . . 

250 

ppm LIVER . . . 

250 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

250 

ppm PINEAL GLAND No Section . . 

250 

ppm PITUITARY Cyst(s) P . 

250 PREOPTIC NUCLEUS . . . 
B-1 

ppm 

250 

ppm TESTIS . . . 

250 

ppm TESTIS Missing one of pair . . 

250 

ppm ADRENAL . . . 

250 

ppm BURSA No Section . . 

250 

ppm CLOACAL GLAND . . . 

250 

ppm KIDNEY . . . 

250 

ppm LIVER . . . 

250 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

250 

ppm PINEAL GLAND . . . 

250 

ppm PITUITARY . . . 

250 

ppm PREOPTIC NUCLEUS . . . 

250 

ppm TESTIS No Section . . 

250 

ppm ADRENAL . . . 

250 

ppm BURSA . . . 

250 

ppm CLOACAL GLAND . . . 

250 

ppm KIDNEY . . . 

250 

ppm LIVER . . . 

250 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

250 

ppm PINEAL GLAND No Section . . 

250 

ppm PITUITARY . . . 

250 

ppm PREOPTIC NUCLEUS . . . 

250 

ppm TESTIS . . . 

250 

ppm TESTIS Missing one of pair . . 

250 

ppm ADRENAL . . . 

250 

ppm BURSA . . . 

250 

ppm CLOACAL GLAND . . . 

250 

ppm KIDNEY . . . 
B-1 

250 

ppm LIVER . . . 

250 

ppm 

MESONEPHRIC/WOLFFIAN 

DUCTS . . . 

250 

ppm PINEAL GLAND . . . 

250 

ppm PITUITARY . . . 

250 

ppm PREOPTIC NUCLEUS . . . 

250 

ppm TESTIS No Section . . 

0 ppm ADRENAL . . . 

0 ppm BRAIN Encephalomalacia, Regionally Extensive 3 3 

0 ppm BURSA . . . 

0 ppm CLOACAL GLAND . . . 

0 ppm KIDNEY . . . 

0 ppm LIVER . . . 

0 ppm MULLERIAN DUCTS . . . 

0 ppm OVARY . . . 

0 ppm PINEAL GLAND No Section . . 

0 ppm PITUITARY . . . 

0 ppm PREOPTIC NUCLEUS . . . 

0 ppm ADRENAL . . . 

0 ppm BURSA . . . 

0 ppm CLOACAL GLAND No Section . . 

0 ppm KIDNEY . . . 

0 ppm LIVER . . . 

0 ppm MULLERIAN DUCTS . . . 

0 ppm OVARY . . . 

0 ppm PINEAL GLAND No Section . . 

0 ppm PITUITARY No Section . . 

0 ppm PREOPTIC NUCLEUS . . . 

0 ppm ADRENAL . . . 

0 ppm BURSA Apoptotic Necrosis, Increased 1 . 

0 ppm CEREBELLUM . . 1 

0 ppm CLOACAL GLAND . . . 

0 ppm KIDNEY . . . 

0 ppm LIVER . . . 

0 ppm MULLERIAN DUCTS No Section . . 

0 ppm OVARY . . . 

0 ppm PINEAL GLAND No Section . . 

0 ppm PITUITARY . . . 

0 ppm PREOPTIC NUCLEUS . . 1 

0 ppm ADRENAL . . . 

0 ppm ADRENAL Missing one of pair . . 

0 ppm BURSA . . . 

0 ppm CLOACAL GLAND . . . 

0 ppm KIDNEY . . . 

0 ppm LIVER . . . 

0 ppm MULLERIAN DUCTS . . . 
B-1 

0 ppm OVARY . . . 

0 ppm PINEAL GLAND No Section . . 

0 ppm PITUITARY . . . 

0 ppm PREOPTIC NUCLEUS . . . 

30 

ppm ADRENAL . . . 

30 

ppm BURSA . . . 

30 

ppm CLOACAL GLAND . . . 

30 

ppm KIDNEY . . . 

30 

ppm LIVER . . . 

30 

ppm MULLERIAN DUCTS . . . 

30 

ppm OVARY . . . 

30 

ppm PINEAL GLAND . . . 

30 

ppm PITUITARY . . . 

30 

ppm PREOPTIC NUCLEUS . . . 

30 

ppm ADRENAL No Section . . 

30 

ppm BURSA . . . 

30 

ppm CLOACAL GLAND . . . 

30 

ppm KIDNEY . . . 

30 

ppm LIVER . . . 

30 

ppm MULLERIAN DUCTS . . . 

30 

ppm OVARY No Section . . 

30 

ppm PINEAL GLAND . . . 

30 

ppm PITUITARY Cyst(s) P . 

30 

ppm PREOPTIC NUCLEUS . . . 

30 

ppm ADRENAL . . . 

30 

ppm BURSA . . . 

30 

ppm CLOACAL GLAND . . . 

30 

ppm KIDNEY . . . 

30 

ppm LIVER . . . 

30 MULLERIAN DUCTS . . . 
B-1 

ppm 

30 

ppm OVARY . . . 

30 

ppm PINEAL GLAND No Section . . 

30 

ppm PITUITARY . . . 

30 

ppm PREOPTIC NUCLEUS . . . 

30 

ppm ADRENAL . . . 

30 

ppm BURSA No Section . . 

30 

ppm CLOACAL GLAND . . . 

30 

ppm KIDNEY . . . 

30 

ppm LIVER . . . 

30 

ppm MULLERIAN DUCTS . . . 

30 

ppm OVARY . . . 

30 

ppm PINEAL GLAND No Section . . 

30 

ppm PITUITARY . . . 

30 

ppm PREOPTIC NUCLEUS . . . 

30 

ppm ADRENAL . . . 

30 

ppm BURSA . . . 

30 

ppm CLOACAL GLAND . . . 

30 

ppm KIDNEY . . . 

30 

ppm LIVER . . . 

30 

ppm MULLERIAN DUCTS No Section . . 

30 

ppm OVARY . . . 

30 

ppm PINEAL GLAND No Section . . 

30 

ppm PITUITARY . . . 

30 

ppm PREOPTIC NUCLEUS . . . 

30 

ppm ADRENAL . . . 

30 

ppm BURSA . . . 

30 

ppm CLOACAL GLAND . . . 
B-1 

30 

ppm KIDNEY . . . 

30 

ppm LIVER . . . 

30 

ppm MULLERIAN DUCTS . . . 

30 

ppm OVARY . . . 

30 

ppm PINEAL GLAND No Section . . 

30 

ppm PITUITARY No Section . . 

30 

ppm PREOPTIC NUCLEUS . . . 

30 

ppm ADRENAL . . . 

30 

ppm BURSA . . . 

30 

ppm CLOACAL GLAND . . . 

30 

ppm KIDNEY . . . 

30 

ppm LIVER . . . 

30 

ppm MULLERIAN DUCTS . . . 

30 

ppm OVARY . . . 

30 

ppm PINEAL GLAND No Section . . 

30 

ppm PITUITARY . . . 

30 

ppm PREOPTIC NUCLEUS . . . 

30 

ppm ADRENAL . . . 

30 

ppm BURSA . . . 

30 

ppm CLOACAL GLAND . . . 

30 

ppm KIDNEY . . . 

30 

ppm LIVER Hepatocellular Necrosis, Focally Extensive 1 . 

30 

ppm MULLERIAN DUCTS . . . 

30 

ppm OVARY . . . 

30 

ppm PINEAL GLAND No Section . . 

30 

ppm PITUITARY . . . 

30 

ppm PREOPTIC NUCLEUS . . . 

30 ADRENAL . . . 
B-1 

ppm 

30 

ppm BURSA Apoptotic Necrosis, Increased 1 . 

30 

ppm CLOACAL GLAND . . . 

30 

ppm KIDNEY . . . 

30 

ppm LIVER . . . 

30 

ppm MULLERIAN DUCTS . . . 

30 

ppm OVARY . . . 

30 

ppm PINEAL GLAND No Section . . 

30 

ppm PITUITARY . . . 

30 

ppm PREOPTIC NUCLEUS . . . 

30 

ppm ADRENAL . . . 

30 

ppm BURSA . . . 

30 

ppm CLOACAL GLAND . . . 

30 

ppm KIDNEY . . . 

30 

ppm LIVER . . . 

30 

ppm MULLERIAN DUCTS . . . 

30 

ppm OVARY . . . 

30 

ppm PINEAL GLAND . . . 

30 

ppm PITUITARY . . . 

30 

ppm PREOPTIC NUCLEUS . . . 

30 

ppm ADRENAL . . . 

30 

ppm BURSA Apoptotic Necrosis, Increased 1 . 

30 

ppm CLOACAL GLAND . . . 

30 

ppm KIDNEY . . . 

30 

ppm LIVER . . . 

30 

ppm MULLERIAN DUCTS . . . 

30 

ppm OVARY . . . 

30 

ppm PINEAL GLAND No Section . . 
B-1 

30 

ppm PITUITARY . . . 

30 

ppm PREOPTIC NUCLEUS . . . 

30 

ppm ADRENAL . . . 

30 

ppm BURSA . . . 

30 

ppm CLOACAL GLAND . . . 

30 

ppm KIDNEY . . . 

30 

ppm LIVER . . . 

30 

ppm MULLERIAN DUCTS No Section . . 

30 

ppm OVARY . . . 

30 

ppm PINEAL GLAND . . . 

30 

ppm PITUITARY Cyst(s) P . 

30 

ppm PREOPTIC NUCLEUS . . . 

60 

ppm ADRENAL . . . 

60 

ppm BURSA . . . 

60 

ppm CLOACAL GLAND . . . 

60 

ppm KIDNEY . . . 

60 

ppm LIVER . . . 

60 

ppm MULLERIAN DUCTS . . . 

60 

ppm OVARY . . . 

60 

ppm PINEAL GLAND . . . 

60 

ppm PITUITARY . . . 

60 

ppm PREOPTIC NUCLEUS . . . 

60 

ppm ADRENAL . . . 

60 

ppm BURSA Apoptotic Necrosis, Increased 1 . 

60 

ppm CLOACAL GLAND . . . 

60 

ppm KIDNEY . . . 

60 

ppm LIVER . . . 

60 MULLERIAN DUCTS . . . 
B-1 

ppm 

60 

ppm OVARY . . . 

60 

ppm PINEAL GLAND No Section . . 

60 

ppm PITUITARY . . . 

60 

ppm PREOPTIC NUCLEUS . . . 

60 

ppm ADRENAL . . . 

60 

ppm BURSA . . . 

60 

ppm CLOACAL GLAND . . . 

60 

ppm KIDNEY . . . 

60 

ppm LIVER . . . 

60 

ppm MULLERIAN DUCTS . . . 

60 

ppm OVARY . . . 

60 

ppm PINEAL GLAND No Section . . 

60 

ppm PITUITARY . . . 

60 

ppm PREOPTIC NUCLEUS . . . 

60 

ppm ADRENAL . . . 

60 

ppm BURSA . . . 

60 

ppm CLOACAL GLAND . . . 

60 

ppm KIDNEY . . . 

60 

ppm LIVER . . . 

60 

ppm MULLERIAN DUCTS . . . 

60 

ppm OVARY . . . 

60 

ppm PINEAL GLAND . . . 

60 

ppm PITUITARY . . . 

60 

ppm PREOPTIC NUCLEUS . . . 

60 

ppm ADRENAL . . . 

60 

ppm BURSA . . . 

60 

ppm CLOACAL GLAND . . . 
B-1 

60 

ppm KIDNEY . . . 

60 

ppm LIVER . . . 

60 

ppm MULLERIAN DUCTS . . . 

60 

ppm OVARY . . . 

60 

ppm PINEAL GLAND No Section . . 

60 

ppm PITUITARY . . . 

60 

ppm PREOPTIC NUCLEUS . . . 

60 

ppm ADRENAL . . . 

60 

ppm BURSA . . . 

60 

ppm CLOACAL GLAND . . . 

60 

ppm KIDNEY . . . 

60 

ppm LIVER . . . 

60 

ppm MULLERIAN DUCTS . . . 

60 

ppm OVARY . . . 

60 

ppm PINEAL GLAND . . . 

60 

ppm PITUITARY . . . 

60 

ppm PREOPTIC NUCLEUS . . . 

60 

ppm ADRENAL . . . 

60 

ppm BURSA . . . 

60 

ppm CLOACAL GLAND . . . 

60 

ppm KIDNEY . . . 

60 

ppm LIVER . . . 

60 

ppm MULLERIAN DUCTS . . . 

60 

ppm OVARY . . . 

60 

ppm PINEAL GLAND No Section . . 

60 

ppm PITUITARY . . . 

60 

ppm PREOPTIC NUCLEUS . . . 

60 ADRENAL . . . 
B-1 

ppm 

60 

ppm BURSA . . . 

60 

ppm CLOACAL GLAND . . . 

60 

ppm KIDNEY . . . 

60 

ppm LIVER . . . 

60 

ppm MULLERIAN DUCTS . . . 

60 

ppm OVARY . . . 

60 

ppm PINEAL GLAND No Section . . 

60 

ppm PITUITARY . . . 

60 

ppm PREOPTIC NUCLEUS . . . 

60 

ppm ADRENAL . . . 

60 

ppm BURSA . . . 

60 

ppm CLOACAL GLAND . . . 

60 

ppm KIDNEY . . . 

60 

ppm LIVER . . . 

60 

ppm MULLERIAN DUCTS No Section . . 

60 

ppm OVARY . . . 

60 

ppm PINEAL GLAND No Section . . 

60 

ppm PITUITARY . . . 

60 

ppm PREOPTIC NUCLEUS . . . 

60 

ppm ADRENAL . . . 

60 

ppm BURSA . . . 

60 

ppm CLOACAL GLAND . . . 

60 

ppm KIDNEY . . . 

60 

ppm LIVER . . . 

60 

ppm MULLERIAN DUCTS . . . 

60 

ppm OVARY . . . 

60 

ppm PINEAL GLAND No Section . . 
B-1 

60 

ppm PITUITARY Cyst(s) P . 

60 

ppm PREOPTIC NUCLEUS . . . 

60 

ppm ADRENAL . . . 

60 

ppm BURSA . . . 

60 

ppm CLOACAL GLAND . . . 

60 

ppm KIDNEY . . . 

60 

ppm LIVER . . . 

60 

ppm MULLERIAN DUCTS . . . 

60 

ppm OVARY . . . 

60 

ppm PINEAL GLAND . . . 

60 

ppm PITUITARY . . . 

60 

ppm PREOPTIC NUCLEUS . . . 

60 

ppm ADRENAL . . . 

60 

ppm BURSA . . . 

60 

ppm CLOACAL GLAND . . . 

60 

ppm KIDNEY . . . 

60 

ppm LIVER . . . 

60 

ppm MULLERIAN DUCTS No Section . . 

60 

ppm OVARY . . . 

60 

ppm PINEAL GLAND . . . 

60 

ppm PITUITARY . . . 

60 

ppm PREOPTIC NUCLEUS . . . 

60 

ppm ADRENAL . . . 

60 

ppm BURSA . . . 

60 

ppm CLOACAL GLAND . . . 

60 

ppm KIDNEY . . . 

60 

ppm LIVER . . . 

60 MULLERIAN DUCTS . . . 
B-1 

ppm 

60 

ppm OVARY . . . 

60 

ppm PINEAL GLAND . . . 

60 

ppm PITUITARY No Section . . 

60 

ppm PREOPTIC NUCLEUS . . . 

60 

ppm ADRENAL . . . 

60 

ppm BURSA . . . 

60 

ppm CLOACAL GLAND . . . 

60 

ppm KIDNEY . . . 

60 

ppm LIVER Hepatocellular Necrosis, Focally Extensive 2 2 

60 

ppm MULLERIAN DUCTS . . . 

60 

ppm OVARY . . . 

60 

ppm PINEAL GLAND . . . 

60 

ppm PITUITARY . . . 

60 

ppm PREOPTIC NUCLEUS . . . 

60 

ppm ADRENAL . . . 

60 

ppm BURSA . . . 

60 

ppm CLOACAL GLAND . . . 

60 

ppm KIDNEY . . . 

60 

ppm LIVER . . . 

60 

ppm MULLERIAN DUCTS No Section . . 

60 

ppm OVARY . . . 

60 

ppm PINEAL GLAND No Section . . 

60 

ppm PITUITARY . . . 

60 

ppm PREOPTIC NUCLEUS . . . 

60 

ppm ADRENAL . . . 

60 

ppm BURSA . . . 

60 

ppm CLOACAL GLAND . . . 
B-1 

60 

ppm KIDNEY . . . 

60 

ppm LIVER Hepatocellular Necrosis, Focally Extensive 1 . 

60 

ppm MULLERIAN DUCTS . . . 

60 

ppm OVARY . . . 

60 

ppm PINEAL GLAND . . . 

60 

ppm PITUITARY . . . 

60 

ppm PREOPTIC NUCLEUS . . . 

125 

ppm ADRENAL . . . 

125 

ppm BURSA No Section . . 

125 

ppm CLOACAL GLAND No Section . . 

125 

ppm KIDNEY . . . 

125 

ppm LIVER . . . 

125 

ppm MULLERIAN DUCTS . . . 

125 

ppm OVARY . . . 

125 

ppm PINEAL GLAND . . . 

125 

ppm PITUITARY . . . 

125 

ppm PREOPTIC NUCLEUS . . . 

125 

ppm ADRENAL . . . 

125 

ppm BURSA Apoptotic Necrosis, Increased 1 . 

125 

ppm CLOACAL GLAND . . . 

125 

ppm KIDNEY . . . 

125 

ppm LIVER . . . 

125 

ppm MULLERIAN DUCTS . . . 

125 

ppm OVARY . . . 

125 

ppm PINEAL GLAND . . . 

125 

ppm PITUITARY . . . 

125 

ppm PREOPTIC NUCLEUS . . . 

125 ADRENAL . . . 
B-1 

ppm 

125 

ppm BURSA Apoptotic Necrosis, Increased 1 . 

125 

ppm CLOACAL GLAND . . . 

125 

ppm KIDNEY . . . 

125 

ppm LIVER Hepatocellular Necrosis, Focally Extensive 2 2 

125 

ppm MULLERIAN DUCTS . . . 

125 

ppm OVARY . . . 

125 

ppm PINEAL GLAND . . . 

125 

ppm PITUITARY . . . 

125 

ppm PREOPTIC NUCLEUS . . . 

125 

ppm ADRENAL . . . 

125 

ppm BURSA Apoptotic Necrosis, Increased 1 . 

125 

ppm CLOACAL GLAND . . . 

125 

ppm KIDNEY . . . 

125 

ppm LIVER . . . 

125 

ppm MULLERIAN DUCTS . . . 

125 

ppm OVARY . . . 

125 

ppm PINEAL GLAND . . . 

125 

ppm PITUITARY . . . 

125 

ppm PREOPTIC NUCLEUS . . . 

250 

ppm ADRENAL No Section . . 

250 

ppm BURSA . . . 

250 

ppm CLOACAL GLAND . . . 

250 

ppm KIDNEY Mineralization 1 . 

250 

ppm LIVER . . . 

250 

ppm MULLERIAN DUCTS . . . 

250 

ppm OVARY . . . 

250 

ppm PINEAL GLAND . . . 
B-1 

250 

ppm PITUITARY Cyst(s) P . 

250 

ppm PREOPTIC NUCLEUS . . . 

250 

ppm ADRENAL No Section . . 

250 

ppm BURSA Apoptotic Necrosis, Increased 2 . 

250 

ppm CLOACAL GLAND . . . 

250 

ppm KIDNEY . . . 

250 

ppm LIVER Bile Duct Hyperplasia 1 1 250 

ppm LIVER Fibrosis, Peribiliary 1 

250 

ppm MULLERIAN DUCTS . . . 

250 

ppm OVARY . . . 

250 

ppm PINEAL GLAND No Section . . 

250 

ppm PITUITARY . . . 

250 

ppm PREOPTIC NUCLEUS . . . 

250 

ppm ADRENAL . . . 

250 

ppm BURSA Apoptotic Necrosis, Increased 1 . 

250 

ppm CLOACAL GLAND . . . 

250 

ppm KIDNEY . . . 

250 

ppm LIVER . . . 

250 

ppm MULLERIAN DUCTS . . . 

250 

ppm OVARY . . . 

250 

ppm PINEAL GLAND . . . 

250 

ppm PITUITARY Cyst(s) P . 

250 

ppm PREOPTIC NUCLEUS . . . 

250 

ppm ADRENAL . . . 

250 

ppm BURSA . . . 

250 

ppm CLOACAL GLAND . . . 

250 

ppm KIDNEY . . . 

250 LIVER . . . 
B-1 

ppm 

250 

ppm MULLERIAN DUCTS . . . 

250 

ppm OVARY . . . 

250 

ppm PINEAL GLAND . . . 

250 

ppm PITUITARY . . . 

250 

ppm PREOPTIC NUCLEUS . . . 

250 

ppm ADRENAL . . . 

250 

ppm BURSA . . . 

250 

ppm CLOACAL GLAND . . . 

250 

ppm KIDNEY . . . 

250 

ppm LIVER . . . 

250 

ppm MULLERIAN DUCTS . . . 

250 

ppm OVARY . . . 

250 

ppm PINEAL GLAND . . . 

250 

ppm PITUITARY . . . 

250 

ppm PREOPTIC NUCLEUS . . . 

250 

ppm ADRENAL . . . 

250 

ppm BURSA . . . 

250 

ppm CLOACAL GLAND . . . 

250 

ppm KIDNEY . . . 

250 

ppm LIVER . . . 

250 

ppm MULLERIAN DUCTS . . . 

250 

ppm OVARY . . . 

250 

ppm PINEAL GLAND . . . 

250 

ppm PITUITARY . . . 

250 

ppm PREOPTIC NUCLEUS . . . 

250 

ppm ADRENAL . . . 

250 

ppm BURSA . . . 
B-1 

250 

ppm CLOACAL GLAND . . . 

250 

ppm KIDNEY . . . 

250 

ppm LIVER . . . 

250 

ppm MULLERIAN DUCTS . . . 

250 

ppm OVARY . . . 

250 

ppm PINEAL GLAND . . . 

250 

ppm PITUITARY . . . 

250 

ppm PREOPTIC NUCLEUS . . . 

250 

ppm ADRENAL . . . 

250 

ppm BURSA . . . 

250 

ppm CLOACAL GLAND . . . 

250 

ppm KIDNEY . . . 

250 

ppm LIVER . . . 

250 

ppm MULLERIAN DUCTS . . . 

250 

ppm OVARY Retained Right Ovary P 1 

250 

ppm PINEAL GLAND No Section . . 

250 

ppm PITUITARY . . . 

250 

ppm PREOPTIC NUCLEUS . . . 

250 

ppm ADRENAL . . . 

250 

ppm BURSA . . . 

250 

ppm CLOACAL GLAND . . . 

250 

ppm KIDNEY . . . 

250 

ppm LIVER . . . 

250 

ppm MULLERIAN DUCTS . . . 

250 

ppm OVARY . . . 

250 

ppm PINEAL GLAND . . . 

250 

ppm PITUITARY No Section . . 

250 PREOPTIC NUCLEUS . . . 
B-1 

ppm 
APPENDIX I 

HORMONE ANALYSES METHODS 
B-1 

UNIVERSITY OF MARYLAND 

DEPARTMENT OF ANIMAL AND AVIAN SCIENCES 

COLLEGE PARK, MD 

DRAFT JAPANESE QUAIL HORMONE ANALYSIS METHODS 
B-1 

Table of Contents 

1.0 Introduction 

2.0 Quality Assurance 

3.0 Execution of the Technical Directive 

4.0 Validation Methods for Avian Samples and Biological Media 

5.0 Hormone Assays Selected 

5.1 Selected Assays and Vender Information 

6.0 Validation of Each Assay 

6.1 Estradiol 

6.1.1 Extraction 

6.1.2 Plasma Samples 

6.1.3 Yolk Samples 

6.1.4 Fecal Samples 

6.2 Testosterone 

6.2.1 Extraction 

6.2.2 Plasma Samples 

6.2.3 Yolk Samples 

6.2.4 Fecal Samples 

6.3 Thyroid Hormones 

6.3.1 Plasma Samples 

6.3.2 Thyroid Gland Samples 

6.3.2.1 Extraction 

7.0 Sample Collection SOPs 

7.1 Plasma Samples 

7.2 Thyroid Gland Samples 

7.3 Egg Yolk Samples 

7.4 Fecal Samples 
B-1 

1.0 Introduction 

The purpose of this Technical Directive 1 (TD1) for Task Order 38 was to validate assay 

methods in order to accurately conduct selected hormone measurements associated with avian 

studies of toxicants. Since many of the environmental chemicals currently of concern are 

hormonally active or endocrine disrupting chemicals, it is critical to measure pertinent endpoints 

to ascertain effects. These endpoints include hormone assays that reflect impacts on 

reproductive, metabolic, and stress responsive endocrine systems. Specific hormone assays 

validated in TD1 included thyroid hormones (T3 and T4), estradiol and testosterone that occur in 

various biological media (e.g. fecal/urate material, blood, egg yolk, and/or thyroid gland). TD1 

required development of reliable assay methods and subsequent determination of concentrations 

for selected hormones in the various media mentioned above collected from the Tier 2 Japanese 

quail two-generation tests. 

TD1 required validation of these hormone analyses for sensitivity, accuracy, precision, and 

specificity to deliver reliable endocrine data in support of the two-generation tests that are 

underway at other sites. Sample preparation must support the requirements of the selected assay 

systems, with extraction methods to remove any interfering compounds from the extract prior to 

assay. Proper extraction will reduce variability and error due to interference. Sample volumes 

or weights must be sufficient to ensure measureable levels of the hormone of interest. As 

described below, assays were tested for sensitivity and to assure serial dilutions of unknown 

samples behaved in manner parallel to hormone standards (i.e. parallelism). Samples used to 

determine sensitivity and parallelism were either collected by the testing laboratories conducting 

the multi-generation tests or from samples collected from our Japanese quail, which are of a 

similar genetic stock. 

2.0 Quality Assurance

The University of Maryland Quality Assurance (QA) personnel has been informed about 

progress of the validation studies and involved in resolving some issues that arose in working 

through the varied assay systems and extraction procedures. She has been provided with the data 

set and conducted QA using the raw data. Based on her assessment, there will be specifications 

in the Standard Operating Procedures (SOPs) for the use of the validated assays within 

prescribed limits to ensure high quality analyses and accurate data. She will prepare a brief audit 

report that reviews the data and use of appropriate methods. Both the summary report and QA 

report will be reviewed by Battelle QA. 

3.0 Execution of the Technical Directive 

A number of commercially available assay kits were tested to select the best-suited assay 

systems for the samples generated in the multi-generation avian test. Assays have been validated 

for each hormone and in the various matrices. We recommend specific assays on the basis of 

assay performance with hormone concentrations as predicted from our test samples. These 

recommendations consider both the utility of specific analyses for the purpose of screening the 

impact of test substances on endocrine activity. In addition, these assays will be critical in 

assessing individual endocrine responses and characterizing adverse consequences of exposure. 

Data reports and supporting digital files including spreadsheets are provided as both hard copy 
B-1 

and digital electronic formats. The digital information will be recorded on a DVD and delivered 

to the EPA TOPO. The narrative report includes supporting tables and refers to a series of Excel 

spreadsheets for each type of assay that contains the complete description of the assay validation, 

with reproducibility and detection limits for each hormone in each biological matrix. 

A fundamental decision was made regarding the primary assay methodology using available 

commercial assay kits. This decision was whether to use Enzyme Linked Immunoassays 

(ELISA) or a radioimmunoassay (RIA). Commercial assays were tested to determine if ELISA 

or RIA provides the most advantageous system for analysis of a particular hormone. The assays 

tested were those that we have either used or those that have been recently developed for use 

with bird samples. An assay system was selected based on the performance capabilities, 

including sensitivity, validity, and reliability. Once selected, each assay system was further 

tested with avian samples to ascertain the working limits for that assay as well as effects of the 

matrix within which the hormone resides. These working limits have been incorporated into the 

SOPs developed for each assay system to allow for maximal reliability and comparability across 

experiments and studies. 

4.0 Validation Methods for Avian Samples and Biological Media 

Several sources of samples were used to complete the initial testing and subsequent validation of 

the assays. For initial testing of the efficacy of assay systems, pools of chicken serum or plasma 

were used to establish basic working performance of each assay. This allowed for comparison of 

the span of the standard curves and establishment of the limits of sensitivity as well as collecting 

information on the repeatability of the within assay measurements for the points on the standard 

curve. Assay of extracted chicken serum or plasma provided information about the accuracy, 

precision, and sensitivity of each assay for use with an avian sample and if there were any 

differences in the measurement of serum or plasma samples. Once the number of potential assay 

systems was reduced, then quail samples were used to test the assay systems. These quail 

samples were from the University of Maryland Japanese quail colony; they were archived, frozen 

samples collected from adults and hatchlings. These samples provided insight into the working 

limits of sensitivity of the assays and allowed us to select assays that would be likely to detect 

samples well within the working range of the standard curve for each hormone. In addition, 

thyroid, egg yolk and fecal samples from the University of Maryland quail colony were used to 

provide pooled samples to test the assays in this other matrix. 

Two sets of samples were received from the EPA. The first set was intended for assay validation 

and included thyroid glands, serum, and plasma from Bayer Crop Sciences. The blood samples 

from this set were entirely unusable due to poor quality. The plasma samples consisted of what 

appeared to be packed red blood cells, and the serum samples showed unacceptable levels of 

hemolysis, which interferes with the antibody-antigen interactions in both ELISAs and RIAs. 

The thyroid glands from this set of samples are being used to validate extraction procedures. 

A second set of approximately 300 quail samples were received from Smithers Viscient (labeled 

TP35) sent by Carol Sloan (RTI). They included F0 adult (175 sample tubes) and embryo 

plasma samples (~94 samples), collected using heparin as an anticoagulant. The adult samples 

were visually inspected for quality and quantity. Of the 175 sample tubes, most had ample 
B-1 

volume to run all the assays with some surplus and appeared to be of good quality. Seventeen of 

the adult samples contained less than the necessary volume (see below), and 5 of the samples 

were severely hemolyzed. It was been noted which samples were inadequate in volume and/or 

quality. Embryo samples will have to be pooled, as the volume in most of the samples appears to 

be approximately 100μl. 

Each matrix was separately validated for use with each assay to ensure performance standards. 

This meant that samples from blood (plasma), thyroid gland, feces, and yolk were each be tested 

and validated according to the appropriate assay system. In addition, samples from different life 

stages (adult and hatchling) were tested to ascertain stability of the assay and sufficient detection 

sensitivity. Finally, sensitivity was determined for each assay system relative to the predicted 

sample volume necessary to achieve the threshold concentration for detection. 

Each assay system was tested for the following criteria: sensitivity, specificity, accuracy, and 

precision. The validation criteria are described below. These criteria were applied to the extracts 

from the different media, including serum/plasma, thyroid gland, feces, and yolk. 

Sensitivity is ability of the assay to distinguish between two similar concentrations. It is 

determined by testing the standard deviation of 10 replicate measurements at each of three 

selected concentrations. The variability determines the sensitivity of measurements. This 

threshold of variability is expressed according to pg/ml hormone concentration, which denotes 

the sensitivity of the assay. Also, the test of parallelism provides additional information about 

the volume requirements for each type of sample. 

Specificity is defined as the capability to discern a particular hormone as distinct from other 

species of hormones that have similar structure. This characteristic is generally determined by 

the company that produces the assay kit by comparison of antibody binding to a range of other 

hormones that are likely to be cross reactive with the antibody. In the case of testosterone, it is 

unusual to have an assay with an antibody showing no cross reactivity with 5dihydrotestosterone (5-DHT). As such, most assays for androgenic steroids are termed androgen 

assays and this generally denotes a 100% cross reactivity with testosterone and an accompanying 

50% cross reactivity with 5-DHT. We did not test the cross reactivity for similar hormones 

and relied on the manufacturers’ quality assurance and product information. 

In addition, a test of parallelism was conducted to ensure that there was no interference due to 

compounds present in the extract. Parallelism was conducted using a serum/plasma pool extract 

that is serially diluted and assayed. There were at least 3 series conducted and then averaged to 

see where the measured concentrations fall on the curve. Additionally, the curve produced by 

the serial dilution should parallel the standard curve. These determinations also provide an 

indication of limits of sensitivity relative to sample volume. The slopes were compared to 

determine if there is a significant difference in the parallel curves. 

Accuracy denotes the closeness of measured concentrations of a hormone with the predicted 

concentration as measured by the standard curve. The accuracy of an assay was determined by 

replicate assays of three selected standards (n=6/standard concentration). The error of 

measurements did not exceed 10% of the standard concentration. 
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Precision is the repeatability of measurements by an assay. There is intra-assay variation and 

inter-assay variation. Three points on the standard curve were selected as targets for pooled 

internal controls and analyzed in each assay. The Coefficient of Variation around each measured 

concentration was less than 10%. 

Extraction Methods differed according to the media. Routine extraction methods were tested 

for recovery of added hormone to ensure efficiency of the process. Samples were extracted in a 

manner to optimize the reliability of consistent and high percentage recovery. Yolk samples 

were tested for the most efficient extraction method and several approaches were tested to 

determine if fractional separation was necessary. A standardized extraction method was 

finalized for each matrix for each assay as described below and assay validation was performed 

using all types of extracts. 

5.0 Hormone Assays Selected 

5.1 Selected Assays and Vender Information 

Assay kits recommended for use in the Avian Two-Generation Studies are listed in Table 1. 

This table has supporting information below that specifies the criteria that were used in the 

selection of each kit. One of the primary issues was that of using an Enzyme Linked Immuno 

Sorbent Assay (ELISA) or radioimmunoassay (RIA). We recommend RIAs for most of the 

hormones, given sensitivity and sample volume constraints. Additionally, the RIA is more 

efficient in terms of measurement of large numbers of samples. See bulleted points below for 

more detailed explanation. 

Table 1. Assay kits evaluated for measurement of triiodothyronine (T3), thyroxine (T4), estradiol 

(E2), and testosterone (T) in avian serum or plasma samples. 

Hormone Assay 

Type 

Company Catalog # Reported 

Sensitivity 

Required Volume 

(μl)1

T3 

Total ELISA Calbiotech T3104T 0.5 ng/ml 100 

 RIA MP 

Biomedicals 

06B254215 0.5 ng/ml 200 

 RIA2 Siemens TKT31 0.2 ng/ml 200 

Free RIA Siemens TKF31 0.5 pg/ml 200 

T4 

Total ELISA Calbiotech T4105T 20 ng/ml 50 

 RIA MP 

Biomedicals 

06B254011 20 ng/ml 50 

 RIA2 Siemens TKT41 10 ng/ml 50 

Free RIA Siemens TKF41 1 pg/ml 100 

E2 ELISA Caymen 582251 6.6 pg/ml 100 

 RIA MP 

Biomedicals 

7238102 10 pg/ml 200 
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 RIA2 Siemens TKE21 20 pg/ml 200 

T 

Total ELISA2 Caymen 582701 3.9 pg/ml 100 

 RIA MP 

Biomedicals 

7189102 100 pg/ml 100 

Free RIA Siemens TKTF2 0.55 pg/ml 100 

1

For analysis of each sample in duplicate 

2

Kits indicated in bold were selected for final analysis of plasma, thyroid glands, yolk, and feces 

from Japanese quail. 

 The above kits were all evaluated for analysis of hormones in avian (chicken) plasma and 

serum, both as is and extracted with diethyl ether and/or methylene chloride. 

o All kits worked well and yielded linear standard curves within the reported range 

of the assay. 

o It was determined that RIAs are preferred over ELISAs for analysis of large 

numbers of samples due to more simplified methodology with less hands-on time, 

ability to assay several hundred samples at one time, and cost. Unfortunately, 

neither RIA evaluated for measurement of testosterone in avian samples was 

sufficiently sensitive. 

 For total T3 and total T4, Siemens RIAs were selected for final use 

because of their increased sensitivity. Assays for free T3 and free T4 did 

not detect any measurable hormone. 

 Although the reported sensitivity of the estradiol RIA from MP 

Biomedicals is greater than that of the Siemens RIA, the levels for nonspecific binding measured with that assay were unacceptable. Therefore, 

the RIA from Siemens was selected for final use. 

 The testosterone RIA from MP Biomedicals was not sufficiently sensitive 

for avian samples, and the free testosterone RIA from Siemens did not 

detect any measurable hormone. The only assay with sufficient sensitivity 

for measurement of testosterone in avian samples was the Caymen ELISA, 

and it was selected for final use. 

o Initial runs with chicken serum and plasma indicated that hormone measurements 

for total T3, total T4, and estradiol were approaching the limit of sensitivities of 

the assays, and there was a need to attempt to improve assay sensitivity. 

 For hormone measurement in blood samples, plasma is preferred over serum. While both 

worked well in initial evaluations, the volume of recovery from whole blood is greater in 

plasma than in serum. Preparation of plasma using either heparin or EDTA as the 

anticoagulant was compatible with the assays, though almost all of the validation was 

completed in plasma prepared with heparin as the anticoagulant so this would be the 

preferred method for plasma collection. 
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 For analysis of all hormones, a minimum volume of 600 μl plasma is required. Of 

this, 350 μl must be extracted for measurement of steroid hormones and 250 μl is 

needed for measurement of thyroid hormones in unextracted plasma. 

 Thyroid hormones are lost during organic extraction and must be evaluated in 

unextracted serum or plasma. 

 Initial results for measurement of estradiol and testosterone indicated that serum and 

plasma must be organically extracted because of a lack of parallelism in unextracted 

samples. 

o Diethyl ether proved more suitable for extraction of steroid hormones from serum 

or plasma, due to ease of extraction and hormone recovery. 

o Extraction efficiencies are being conducted for plasma, yolk, and feces by spiking 

samples with tritiated estradiol or tritiated testosterone. 

After initial evaluation of both RIAs and ELISAs from several companies, the kits indicated in 

bold in Table 1 were selected for final use (see above for rationale). All selected kits were for 

evaluated for specificity, intra- and inter-assay variation, and parallelism in sample types to be 

measured in Japanese quail. The RIAs for T3, T4, and E2 were also evaluated for the ability to 

improve assay sensitivity. Assay characteristics are shown in Table 2. Demonstration of 

parallelism is shown on the graphs. 

 Limits of sensitivity were determined by running a series of dilutions until the % of 

maximum binding [(B/B0)*100] was at least 95% and repeatable in multiple assays. 

Actual limits of sensitivity for T3, T4, and E2 were improved approximately 10-fold 

(compare Table 2 vs Table 1). When performing these assays, the standard curve is 

adjusted accordingly. 

 Specificity was determined by measuring a known amount of the indicated hormone in 

the assay (i.e. T4 in the T3 assay, T3 in the T4 assay, T in the E2 assay, and E2 in the T 

assay), and determining cross-reactivity (%) as [(amount measured/amount added)*100]. 

 Intra- and inter-assay variation (coefficient of variation (CV; %) were determined in all 

four assays using chicken plasma because of the large sample volume required. In each of 

6 independent assay runs, the same sample was analyzed 6 times. The intra-assay CV was 

determined in each of the runs, and the average of the 6 runs was determined to be the 

intra-assay CV (%). The inter-assay CV (%) was determined from the mean value of the 

sample in each of the six runs. For total T4 and estradiol, the levels in chicken plasma 

approached the limit of sensitivity of the assay and contributed to higher variability. 

Therefore, additional intra-assay and inter-assay values for these assays were determined 

in the same manner using thyroid extract (total T4) and fecal extract (estradiol). 

 Parallelism was evaluated in each of the sample types for each of the assays. At least 4 

dilutions of a given sample were run on a minimum of 3 independent pools of each 

sample type. In addition, 3 independent pools of hatchling plasma were evaluated in each 
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assay at 2 dilutions. This allowed us to further validate the assays in younger birds from a 

different colony. 

o Total T3: adult plasma (male, female, mixed sex), thyroid extracts 

o Total T4: adult plasma (male, female, mixed sex), thyroid extracts 

o E2: ether-extracted plasma (male, female (2), mixed sex), ether-extracted yolk, 

ethanol-extracted feces. 

o T: ether-extracted plasma (male, female, mixed sex), ether-extracted yolk, 

ethanol-extracted feces. 

Table 2. Assay characteristics of kits selected for final analysis of triiodothyronine (T3), 

thyroxine (T4), estradiol (E2), and testosterone (T) in Japanese quail plasma, thyroid glands, 

yolk, and feces. 

Hormon

e 

Assay Determined 

Sensitivity 

Crossreactivity1

(%)

Intra-assay CV 

(%) 

Inter-assay CV 

(%) 

Total T3 Siemens 

TKT31 

0.03 ng/ml 1.4 (T4) Plasma: 3.7 Plasma: 5.9 

Total T4 Siemens 

TKT41 

1.6 ng/ml 3.3 (T3) Plasma: 9.9 

Thyroid2: 2.9 

Plasma: 13.6 

Thyroid2: 15.2 

E2 Siemens 

TKE21 

2.5 pg/ml 0.2 (T) Plasma: 6.9 

Feces2

: 3.7 

Plasma: 13.7 

Feces2: 10.0 

T Caymen 

582701

3.9 pg/ml 0.1 (E2) Plasma: 5.7 Plasma: 6.4 

1

Measured cross-reactivity is with the hormone indicated in parentheses. 

2

Additional intra- and inter-assay CV values were determined in thyroid extracts for total T4 and 

fecal extracts for E2 because plasma measurements were approaching the limitation of sensitivity 

for the assays and thought to contribute to increased variability in these determinations. 

6.0 Validation of Each Assay 

6.1 Estradiol (see Excel spread sheets for detailed information) 

6.1.1 Steroid Extraction from Various Matrices 

Samples were tested for assay efficacy with or without organic chemical extraction; however, 

steroid hormone assays required chemical extraction in order to remove interfering lipids and 

other compounds that interfere with antibody binding. For both the chicken and the quail steroid 

hormone assays, samples were chemically extracted, using ether for the organic solvent. A 

protocol for yolk analysis was developed in which a series of dilutions was been made with 

distilled-deionized water. Because the distribution of steroids in the yolk is not homogeneous, 

but rather is laid down in layers according to the lay down of the yolk, it was critical to have the 

yolk homogenized to have an accurate measure of the steroid content. Adding the water also 

helps to evenly distribute the steroid and prepare a mixture of yolk:water that is more easily 

extracted with higher steroid hormone recovery from the yolk matrix. Using eggs from the 

UMD Japanese quail colony, different amounts of water were tested for making the yolk:water 
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mixture and for extraction efficacy and reconstitution into the assay buffer. The dilutions 

included 1:1, 1:2, 1:3 (yolk:water). Results showed that 1:1 provided reasonable extractability, 

with the 1:2 dilution providing a better extraction matrix with less carry over of lipids during 

extraction. Extracts were tested in an alcohol clean up procedure to remove excess lipid and 

provide an extract that does not interfere with the steroid assays. Fecal samples were extracted 

using a 90:10 ethanol:water procedure (see Appendix 1, 2, and 3 for detailed SOPs) 

6.1.2 Plasma Samples 

6.1.3 Yolk Samples 

6.1.4 Fecal Samples 

6.2 Testosterone (see Excel spread sheet) 

6.2.1 Extraction (see Section 6.1.1) 

6.2.2 Plasma Samples 

6.2.3 Yolk Samples 

6.2.4 Fecal Samples 

6.3 Thyroid Hormones (see attached spread sheet) 

6.3.1 Plasma Samples 

6.3.2 Thyroid Gland Samples (see Appendix 4 for extraction and sample 

preparation SOP) 

 
B-1 

7.0 Sample Collection SOPs 

Recommendations for Preparation of Samples

Plasma collected using either heparin or EDTA both should work well for all the assays (total 

T3, total T4, estradiol and testosterone). Since most of the validation was conducted using quail 

plasma prepared with heparin, this is the preferred anticoagulant for use in the studies. Quail 

plasma should be collected into a 2-ml centrifuge tube that is clearly and legibly labeled with the 

bird identification. Samples should be stored at – 20o

C and shipped on dry ice. A sample 

inventory in the form a spreadsheet, clearly indicating bird identification and approximate 

plasma volume, must be provided in both an electronic and a hard copy. 

The estimated amount of plasma in the embryo samples is 50-150l. Depending on the 

hormone concentrations in these samples, we recommend that samples be pooled in order to run 

all the assays. The volumes required to run samples in duplicate for each assay are T3=200l; 

T4=50l; estradiol=200l; testosterone=100l. The T3 and T4 assays use neat plasma, with no 

extraction. Assuming the samples cannot be diluted to run in the assays, the approximate total 

amount of plasma required to conduct all assays is 600l. 

Thyroid gland samples from adults, hatchling, and embryos should be provided frozen in 2-ml 

round bottom centrifuge tubes clearly and legibly labeled with the bird identification. The 

thyroid glands need to be weighed at collection. If the combined weight of the glands is more 

than 10 mg, a piece between 5-10 mg needs to be accurately weighed and placed in the sample 

tube. The glands should be stored at -20 C and shipped on dry ice. A sample inventory in the 

form of a spreadsheet, clearly indicating bird identification, total thyroid gland weight, and 

weight of the sample collected into the tube needs to be provided in both an electronic and hard 

copy. 

Yolk samples should be collected with clean separation from albumin because albumin may 

interfere with the assay. The entire yolk should be weighed and homogenized with two volumes 

of ultrapure deionized water. A 1-ml aliquot of the homogenized, diluted yolk should be put into 

a 1.7-ml centrifuge tube that is clearly and legibly labeled with the bird identification. Samples 

should be stored at – 20o

C and shipped on dry ice. A sample inventory in the form of a 

spreadsheet, clearly indicating bird identification, total yolk weight, amount of water used for 

dilution, and volume (if other than 1-ml) needs to be provided in both an electronic and a hard 

copy. 

Please see below for detailed SOPs. 

7.1 Plasma Sample Preparation 

1. If collecting during euthanasia, sever the neck and collect trunk blood into a 

polypropylene plastic 12x75 mm tube containing anticoagulant [heparin is preferred (10 

μl per 1.0 ml blood); EDTA is okay too (10 μl 0.5M per ml of blood)]. No more than 3 

ml should be collected per tube. If collecting by venipuncture, draw blood through needle 

into syringe that has been pre-coated with anticoaculant, remove the needle, and expel 

blood into tube. Collect ~2.5X volume of plasma needed. 
B-1 

2. Record the volume of total blood collected. 

3. Immediately invert the blood sample to allow the anticoagulant to mix with the blood. 

4. Centrifuge the blood for 10 min at 1,500xg and 4 o

C. Plasma should be clear or light tan 

in color. Reddish color indicates substantial red blood cell lysis and presence of 

hemoglobin, which interferes with many assays. Opaqueness indicates the presence of 

particulates, which should be centrifuged out. 

5. Transfer the supernatant to a clean 1.7-ml polypropylene microcentrifuge tube. Tubes 

should be clearly and legibly labeled with bird identification. Record the volume of 

plasma and indicate any discoloration and/or opaqueness in each sample. 

6. Plasma samples should be stored at -20o

C and shipped on dry ice. A sample inventory in 

the form of a spreadsheet, clearly indicating bird identification and volume of plasma 

collected must be provided with shipment, in both an electronic and hard copy. 

7.2 Thyroid Gland Sample Preparation 

1. Weigh the entire thyroid gland and record the total weight. 

2. Dissect a small piece of the gland that is between 5-10 mg, weigh the subsample, and 

record the exact subsample weight to the tenth of a milligram. 

3. Place the thyroid subsample into a 2-ml round bottom microcentrifuge tube (e.g. 

Eppendorf catalog # 022363352) on ice. Tubes should be clearly and legibly labeled with 

bird identification. 

4. Thyroid samples should be stored at -20o

C and shipped on dry ice. A sample inventory in 

the form of a spreadsheet, clearly indicating bird identification, total thyroid weight, and 

exact weight of the subsample within the tube, must be provided with shipment, in both 

electronic and hard copy. 

7.3 Egg Yolk Sample Preparation 

1. Cleanly separate the yolk from the albumen. 

2. Weigh the entire yolk and record the yolk weight. 

3. Calculate and record the volume of water so the entire yolk is diluted with 2 volumes of 

ultrapure deionized water (i.e. 2 mls water/g yolk). 

4. Thoroughly homogenize the entire yolk with the appropriate volume of water determined 

in step 3. 
B-1 

5. Transfer 1-ml of the diluted, homogenized yolk into a 1.7-ml polypropylene 

microcentrifuge tube on ice. If the volume is other than 1-ml, record the actual volume 

transferred. Tubes should be clearly and legibly labeled with bird identification. 

6. Yolk samples should be stored at -20o

C and shipped on dry ice. A sample inventory in 

the form of a spreadsheet, clearly indicating bird identification, total yolk weight, amount 

of water used for dilution, and subsample volume (if other than 1-ml) must be provided 

with shipment, in both electronic and hard copy. 

7.4 Fecal Sample Preparation 

1. Thoroughly mix wet feces to ensure a randomly distributed sample, and place 

approximately 0.5 g into a weigh boat. Record the exact wet weight. 

2. Place wet fecal samples into a dessicator for at least 24 hrs. Periodically check the 

weight, and remove from desicator when the sample is no longer losing moisture. Record 

the exact dry weight and determine % dry matter. 

3. Thoroughly mix the dry fecal sample with a mortar and pestle. Weigh approximately 0.2 

g dry fecal sample and record the exact weight. 

4. Place the weighed dry fecal sample into a 15-ml polypropylene screw cap tube. Tubes 

should be clearly and legibly labeled with bird identification. 

5. Fecal samples should be stored at -20o

C and shipped on dry ice. A sample inventory in 

the form of a spreadsheet, clearly indicating bird identification, total fecal wet weight, 

total fecal dry weight, % dry matter, and exact weight of the subsample within the tube, 

must be provided shipment, in both electronic and hard copy. 
B-1 

Appendix 1: Plasma Extraction for Steroid Hormone Measurement 

Reagents

Diethyl ether (Sigma catalog # 346136)

1X EIA Buffer [diluted from 10X EIA buffer (Caymen Chemical catalog # 400060) using 

ultrapure distilled water] 

*Note*: Volumes below are for extraction and measurement of both estradiol and testosterone 

using Siemens RIA (catalog # TKE21) and Caymen ELISA (catalog # 582701). If smaller or 

larger amounts are to be extracted, adjust the volume of ether and reconstitution buffer 

accordingly. The important thing is that plasma samples are extracted twice with 5X volumes of 

ether and reconstituted in 1X volume EIA buffer to provide a consistent volume between the 

original and extracted samples. A maximum of 1 mL can be extracted in the tubes below. If 

much smaller volumes are being extracted, this can be carried out in 12x75 mm glass culture 

tubes (e.g. Fisher catalog # 14-961-26). 

Procedure: 

1. Label two 18x150 mm glass culture tubes (e.g. VWR catalog # 60825-443) for each 

sample. 

2. Pipet 400 μl plasma into one of the labeled tubes and cover with foil. 

3. Heat plasma in 65oC water bath for 1 hr to bring steroids off the glass and partially 

denature proteins. Allow samples to cool to room temperature. 

4. In the fume hood, add 2 mL diethyl ether to each sample and vortex vigorously for 30 

sec. 

5. Cover with foil or parafilm and allow samples to stand for ~2 min until fractions separate. 

6. Place samples in a -80o

C freezer for ~5 min or snap-freeze using a dry ice/ethanol bath 

until the lower aqueous phase freezes. 

7. In the fume hood, decant the ether fraction into a second labeled tube for that sample. 

8. Allow the extracted plasma to thaw. In the fume hood, add 2 mL diethyl ether to each 

sample and vortex vigorously for 30 sec. 

9. Repeat steps 5 through 7, so that samples are extracted twice and both ether fractions are 

combined into the same tube. 
B-1 

10. Allow samples to stand overnight in fume hood to evaporate. As they are drying, vortex 

periodically to rinse down any steroid that may stick to the side of tube. If necessary, to 

speed evaporation, the samples can be dried in the fume hood under a steady stream of 

nitrogen. 

11. Reconstitute extracted plasma with 400 μl 1X EIA buffer. Vortex for 30 sec and place in 

65o

C water bath for 1 hr. Vortex for 15-20 sec every 15 min. 

12. Allow samples to stand for ~5 min to drain to the bottom of the tube and transfer to a 

clearly and legibly labeled 1.7-mL polypropylene microcentrifuge tube. 

13. Store extracted plasma samples at -20oC. 

 
B-1 

Appendix 2: Yolk Extraction for Steroid Hormone Measurement 

Reagents

Diethyl ether (Sigma catalog # 346136)

1X EIA Buffer [diluted from 10X EIA buffer (Caymen Chemical catalog # 400060) using 

ultrapure distilled water] 

Procedure: 

1. Label two 18x150 mm glass culture tubes (e.g. VWR catalog # 60825-443) for each 

sample. 

2. Thaw and thoroughly vortex yolk sample, which has previously been homogenized with 

2 parts ultrapure distilled water. 

3. Pipet 300 μl yolk sample into one of the labeled tubes. 

4. In the fume hood, add 6 mL diethyl ether to each sample and vortex vigorously for 30 

sec. 

5. Cover with foil or parafilm and allow samples to stand for 30 min until fractions separate. 

Note that the ether phase will remain cloudy, but most of the lipid-rich yolk phase will 

settle out. 

6. Place samples in a -80o

C freezer for ~5 min or snap-freeze using a dry ice/ethanol bath 

until the lower phase freezes. 

7. In the fume hood, decant the ether fraction into a second labeled tube for that sample. 

8. Allow the extracted yolk to thaw. In the fume hood, add 6 mL diethyl ether to each 

sample and vortex vigorously for 30 sec. 

9. Repeat steps 5 through 7, so that samples are extracted twice and both ether fractions are 

combined into the same tube. 

10. Allow samples to stand in fume hood to evaporate (~2 nights). As they are drying, vortex 

periodically to rinse down any steroid that may stick to the side of tube. If necessary, to 

speed evaporation, the samples can be dried in the fume hood under a steady stream of 

nitrogen. 

11. Reconstitute extracted yolk with 600 μl 1X EIA buffer. Vortex for 30 sec and place in 

65o

C water bath for 1 hr. Vortex for 15-20 sec every 15 min. 
B-1 

12. Allow samples to stand for ~5 min to drain to the bottom of the tube and transfer to a 

clearly and legibly labeled 1.7-mL polypropylene microcentrifuge tube. 

13. Store extracted yolk samples at -20o

C. 
B-1 

Appendix 3: Fecal Matter Extraction for Steroid Hormone Measurement 

Reagents

Ultrapure distilled water

100% Ethanol

1X EIA Buffer [diluted from 10X EIA buffer (Caymen Chemical catalog # 400060) using 

ultrapure distilled water] 

Procedure: 

1. Label one 15-mL polypropylene centrifuge tube (VWR catalog # 89004-368) for each 

sample. 

2. Weigh ~0.2 g dried fecal matter into the labeled 15-mL tube. 

3. Add 1 mL of ultrapure water to each sample and vortex. 

4. Add 9 mL 100% ethanol to each sample and vortex vigorously for 30 sec. Be sure the cap 

is secured tightly and wrap with parafilm. 

5. Secure each tube horizontally to a rocker/shaker and rock vigorously overnight at room 

temperature. 

6. Centrifuge samples at 3000 rpm for 25 min (room temperature). 

7. Transfer supernatant to a clean, labeled 50-mL polypropylene tube (VWR catalog # 

89004-364). 

8. Place tubes into a 50-60o

C water bath and evaporate samples under a stream of nitrogen. 

9. Reconstitute extracted fecal samples with 2 mL 1X EIA buffer. Vortex for 30 sec and 

place in 65o

C water bath for 1 hr. Vortex for 15-20 sec every 15 min. 

10. Transfer samples to two clearly and legibly labeled 1.7-mL polypropylene 

microcentrifuge tubes. 

11. Store at extracted fecal samples at -20oC. 
B-1 

Appendix 4: Thyroid Gland Extraction for Thyroid Hormone Measurement 

Reagents 

Digestion Mix (100 ml)

1.21 g TRIS 

153.65 mg L-Glutathione reduced (Sigma catalog # G4251) 

42.55 mg PTU (6-n-propyl-2-thiouracil; Sigma P-3755) 

1ml Triton-X-100 (Fisher catalog # BP151-500) 

Bring to final volume of 100 mL with ultrapure distilled water after stirring to dissolve. 

Store at 4oC. 

Saturated Ammonium Sulfate (SAS)*

76.7 g ammonium sulfate (Fisher catalog # A702500) 

100 mL ultrapure distilled water 

Cover the beaker with foil and stir until dissolved while heating to ~65o

C. 

Place at 4oC overnight to allow precipitation to occur. 

Filter the solution into a new bottle and store at 4oC. Further precipitation may occur upon longterm storage. 

*SAS can also be purchased from Fisher (catalog # PI-45216) 

Pronase (Calbiochem catalog # 53702; can be purchased from Fisher) 

Procedure: 

1. Thyroid gland subsamples weighing ~5 mg should be in a 2-ml roundbottom microfuge 

tube (e.g. Eppendorf catalog # 022363352). Note the exact weight. 

2. Calculate the total volume pronase digestion buffer needed such that each sample gets 

50X volume of a 100 mg/ml solution. For example, a sample weighing 5.2 mg will get 

260 μl. To digest 12 samples weighing exactly 5 mg, 3 mL of digestion buffer containing 

300 mg of pronase is needed (0.25 mL x 12 = 3 mL x 100 mg/mL = 300 mg). Weigh out 

the appropriate amount pronase into the appropriate volume of digestion mix pre-warmed 

to 37o

C and vortex to dissolve. Be sure to account for loss during transfer and make extra 

buffer. 

3. Add 50X volumes of 100 mg/ml pronase digestion buffer made in step 2 pre-warmed to 

37oC to each sample. A piece weighing exactly 5 mg will get 250 μl. Adjust the volume 

for the exact weight. 

4. Homogenize each sample for 30-60 sec, until the tissue is no longer visible. 

5. Place samples in a 37o

C water bath for 1-2 hr, and then transfer them to shaking air 

incubator set to 37o

C for a total of 24 hr. If a 37oC shaking water bath is available, it can 

be used for the full 24 hr. 
B-1 

6. Add one volume of ice-cold SAS solution equal to the volume of pronase digestion mix. 

For example, the 5.2 mg sample will get 260 μl SAS. 

7. Place samples on ice for 30 min. 

8. Centrifuge samples at 20,000xg and 4o

C for 30 min. 

9. Transfer the supernatant to a clean 1.7-mL microcentrifuge tube. 

10. Store extracted thyroid glands at -20o

C. 
APPENDIX J 

CONTRIBUTING SCIENTIST REPORT 

STATISTICAL ANALYSIS OF HORMONE DATA 
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Statistical Analysis on Hormone Date for USAPHC 

Toxicology Protocol Number: 08JG-80-08-04-02 

Two-Generation Test for Endocrine Disruption and Reproductive Toxicity in 

Japanese Quail (Corturnix japonica) using Endosulfan 

Good Laboratory Practice Compliance Statement 

The statistical analysis of the data for the Two-generation test for endocrine disruption 

and reproductive toxicity in Japanese quail (Cortunix japonica) using endosulfan was 

conducted in compliance with Title 40, Code of Federal Regulations (CFR), Part 792, 

Good Laboratory Practice Standards. Statistical analyses were conducted according to 

the procedures outlined in the protocol for this study (Protocol Number 08JG-80-08-04-

02). 

 

 29 January, 2013 

Shane Hall Date 

Statistician 
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I. Analysis Methods 

A two-way Analysis Of Variance (ANOVA) was used to analyze four hormones measured (T3, 

T4, Estradiol, and Testosterone) from two independent three-generation studies that involved 

exposing quails to endosulfan. Dosage of either endosulfan was altered to determine if the 

amount of endosulfan the quails received had an effect on their hormone values. Endosulfan was 

divided into five dose groups (control, 30 ppm, 60 ppm, 125 ppm, and 250 ppm). In addition to 

dosage, both male and female quails were used to determine if gender also had an effect on their 

hormone values. 

The ANOVA model, using each of the four hormones as the dependent variable and dose group 

and gender as the predictors, determined if at least two of the groups within each predictor were 

significantly different from one another (test of main effect). The ANOVA model also 

determined if the effect that a change in dose group had on the hormone was significantly 

impacted by gender of the quails (test of interaction). If the interaction was statistically 

significant, the main effect results for dose group and gender are potentially misleading and must 

be interpreted in light of the interaction. Therefore, if an interaction was significant, p-values for 

the component parts are not given. If the interaction was not statistically significant, and dose 

group was, a Tukey post-hoc test was used to determine which groups were significantly 

different from one another. Because gender only consisted of males and females, the ANOVA 

p-value was sufficient to determine a difference between males and females and additional posthoc tests were not necessary. 

Data was checked to determine if ANOVA assumptions were met. A Levene’s test was used to 

check equality of variances and ANOVA-model residuals were examined via a Q-Q plot to 

check normality. If assumptions were violated, data was log-transformed and assumptions were 

checked once again to see if an ANOVA was suitable. For this data, all hormone measures 

satisfied ANOVA assumptions using either the untransformed or Log-transformed data, so 

additional analysis adjustments were not necessary. 

Individual data points were occasionally labeled as “Not Determined – Insufficient Volume”, 

“Not Detectable”, or “Not Detectable (Below Assay Limits)”. In the first two cases, the blood 

sample for that quail was either not tested or was unsuccessfully tested and was therefore 

counted as missing data for the statistical analysis. However, the data points that were labeled as 

below the assay limits were tested, but returned values too low for the diagnostic machine to 

detect. In this case, limit of detection divided by the square root of two was substituted for the 

missing value. 

SPSS 16.0 was used to execute the statistical analysis summarized in the report below. 

Statistical significance throughout the analysis was defined at α=.05 or 95% confidence level. 

II. Results 

A. F0 Generation 
B-1 

The four hormones were not significantly impacted by Endosulfan in the F0 generation. 

Estradiol and Testosterone were however significantly impacted by gender. Females had a 

significantly greater Estradiol average, and males had a significantly greater Testosterone 

average. 

Table 1. Effects of Endosulfan on F0 Generation Quail Hormones

Gender Dose Group

(ppm)

T3 (ng/ml) T4 (ng/ml)* Estradiol (pg/ml) Testosterone (pg/ml)*

N Mean SE N Mean SE N Mean SE N Mean SE

Female

Control 5 1.62 0.20 5 2.54 0.73 5 43.45 6.56 5 522.5 345.9

30 5 1.37 0.09 4 2.32 0.74 5 46.25 3.19 5 868.2 177.7

60 5 1.20 0.21 4 6.98 1.81 5 34.51 2.61 5 339.0 85.1

125 5 1.35 0.22 4 3.74 1.26 5 35.66 7.33 5 453.3 267.9

250 5 1.25 0.10 4 2.86 1.39 5 45.59 7.47 5 467.3 274.3

Male

Control 5 1.46 0.11 5 2.49 0.68 5 10.89 4.02 5 1986.3 732.4

30 5 1.50 0.09 4 2.50 0.50 5 14.55 8.85 5 791.8 136.2

60 5 1.60 0.12 5 2.34 1.07 5 10.86 2.89 5 909.6 126.1

125 5 1.58 0.14 5 2.71 0.51 5 9.88 1.37 5 1264.3 363.3

250 5 1.50 0.09 5 3.26 0.92 5 6.90 2.42 5 1157.0 297.8

P‐Value

Dose = .820 Dose = .568 Dose = .563 Dose = .792

Gender = .070 Gender = .235 Gender = .000       

(Females > Males)

Gender = .000         

(Males > Females)

* Analyzed using Log‐transformed data due to violations in ANOVA assumptions.      
B-1 

B. F1 Generation 

The four hormones were not significantly impacted by Endosulfan in the F1 generation. 

Estradiol and Testosterone were however significantly impacted by gender. Females had a 

significantly greater Estradiol average, and males had a significantly greater Testosterone 

average. 

Table 2. Effects of Endosulfan on F1 Generation Quail Hormones

Gender

Dose

Group

(ppm)

T3 (ng/ml) T4 (ng/ml)* Estradiol (pg/ml) Testosterone (pg/ml)

N Mean SE N Mean SE N Mean SE N Mean SE

Female

Control 5 1.39 0.20 4 3.11 0.70 5 30.88 8.40 5 301.8 174.3

30 5 1.99 0.27 4 1.99 0.53 5 54.52 19.76 5 551.6 98.1

60 5 1.89 0.31 5 1.78 0.41 5 57.93 8.33 5 299.2 64.0

125 5 1.24 0.19 4 3.20 1.39 5 44.24 9.25 5 583.4 280.3

250 6 1.40 0.16 3 3.24 2.11 6 46.79 3.65 6 450.8 144.1

Male

Control 5 1.60 0.16 5 2.51 0.43 5 10.16 2.24 5 925.8 405.1

30 4 1.65 0.12 3 1.52 0.39 5 9.31 2.22 5 1249.1 437.0

60 5 1.63 0.15 5 2.80 0.58 5 9.23 0.62 5 2114.3 720.8

125 5 1.66 0.16 5 3.21 0.38 5 9.49 1.08 5 3280.4 1241.9

250 4 1.39 0.16 3 2.92 0.92 4 12.88 7.16 4 1693.4 586.0

P‐Value

Dose = .148 Dose = .459 Dose = .546 Dose = .159

Gender = .981 Gender = .607 Gender = .000       

(Females > Males)

Gender = .000         

(Males > Females)

* Analyzed using Log‐transformed data due to violations in ANOVA assumptions.      
B-1 

C. F2 Generation 

In the F2 generation, T3, Estradiol, and Testosterone were all significantly impacted by 

Endosulfan dosage and/or gender. 

 T3 had similar averages across Endosulfan dose groups, but males had a significantly 

greater average compared to females. 

 Estradiol was impacted by gender with females having a greater average within all five 

dose groups; however the pattern of Estradiol averages for the dose groups within each 

gender were severely different. Males had a fairly consistent average across all five dose 

groups, while females had an average that increased from the control to dose 60 and 

severely decreases from dose 60 to dose 250. For females dose 250 had a significantly 

lower mean compared to dose 60, and for males there were no significant differences 

between groups 

 Testosterone was impacted by gender with males having a greater average within all five 

dose groups; however the pattern of testosterone averages for the dose groups within each 

gender were severely different. As Endosulfan dose increased, males had a severely 

decreasing average level of testosterone, while females had a generally increasing 

average level of testosterone. For males dose 125 and dose 250 had significantly lower 

means compared to control, and for females no significant differences between groups 

were observed. 

Table 3. Effects of Endosulfan on F2 Generation Quail Hormones

Gender

Dose

Group

(ppm)

T3 (ng/ml) T4 (ng/ml)* Estradiol (pg/ml) Testosterone (pg/ml)

N Mean SE N Mean SE N Mean SE N Mean SE

Female

Control 5 1.84 0.34 2 2.99 1.57 5 30.12 2.75 5 235.3 72.2

30 5 1.42 0.13 4 1.84 0.71 5 39.30 5.30 5 297.5 45.7

60 4 1.18 0.17 3 2.79 1.66 5 49.86 4.67 5 620.5 229.5

125 3 0.89 0.31 3 4.20 2.46 5 36.27 11.38 5 334.6 214.1

250 5 1.38 0.34 4 3.89 1.45 5 18.83 4.98 5 511.3 279.6

Male

Control 5 1.66 0.09 4 2.69 0.09 5 10.17 1.86 5 2191.4 197.0

30 5 1.66 0.32 5 2.96 0.94 5 6.95 1.53 5 1759.1 425.1

60 5 1.80 0.14 5 1.72 0.24 5 5.61 1.96 5 1143.9 300.1

125 4 1.54 0.20 5 1.95 0.82 5 7.65 1.47 5 899.7 213.8

250 5 1.80 0.25 5 3.39 0.92 5 6.42 2.33 5 785.7 252.6

P‐Value Dose = .388 Dose = .576 Dose * Gender = Dose * Gender =
B-1 

Gender  = .033         

(Males > Females) Gender = .712

.022 .006

* Analyzed using Log‐transformed data due to violations in ANOVA assumptions.      

APPENDIX K 

RAW DATA 
B-1 

P generation bird weights 

DOSE SEX ID 4/20 4/27 5/4 5/11 5/18 5/25 5/30 6/8 6/15 6/22 6/30 7/6 7/13 

0 F 144 91 102 109 118 125 126 126 134 135 120 106 125 130 

0 F 142 100 112 126 134 130 135 133 137 134 129 127 144 146 

0 F 140 120 134 134 134 136 130 139 137 131 147 145 141 

0 F 139 97 105 119 119 121 126 119 125 128 132 134 131 134 

0 F 141 102 115 116 136 128 131 139 138 149 135 153 150 

0 F 143 105 117 128 129 131 134 140 141 143 146 144 146 149 

0 F 137 104 139 130 130 128 130 104 128 133 130 138 141 140 

0 F 135 121 108 139 138 133 147 145 143 149 142 141 146 103 

0 F 133 96 107 126 124 117 128 128 137 133 130 120 134 136 

0 F 134 103 119 

0 F 136 93 116 132 130 137 135 133 140 145 137 149 145 137 

0 F 138 103 119 129 130 125 130 132 140 141 131 119 143 141 

0 F 145 105 105 127 134 134 127 129 130 129 140 137 141 144 

0 F 146 112 128 119 119 118 118 122 130 134 141 124 142 142 

0 F 147 109 102 119 125 120 128 122 131 134 138 129 128 140 

0 F 148 98 109 117 135 137 138 131 133 134 143 124 144 145 

0 F 149 107 114 140 133 142 137 139 134 150 149 145 114 

0 F 150 117 112 133 148 144 145 155 159 154 160 157 153 154 

0 F 151 108 107 117 124 

0 F 152 110 107 110 

30 F 33 106 129 128 118 126 131 128 136 138 138 142 139 140 

30 F 34 104 113 113 120 118 119 129 129 121 133 134 127 136 

30 F 35 109 115 117 120 128 135 135 126 131 134 138 131 137 

30 f 36 101 118 126 107 109 128 127 136 132 140 135 127 143 

30 F 37 117 120 122 124 128 127 138 147 142 141 150 142 134 

30 F 38 108 130 126 130 132 129 137 136 140 139 141 139 137 

30 F 39 107 118 131 126 134 135 138 138 140 137 117 145 146 

30 F 40 117 126 125 134 142 141 135 146 149 139 147 143 142 

30 F 41 109 132 131 132 141 144 139 151 148 139 140 150 149 

30 F 42 131 123 130 129 128 135 140 141 143 142 145 145 146 

30 F 43 110 120 136 141 142 136 145 144 143 140 143 150 148 

30 F 44 106 114 139 137 141 142 133 147 143 142 142 140 145 

30 F 45 109 113 115 114 114 127 132 116 133 134 141 141 141 

30 F 46 110 115 117 118 117 124 144 144 143 142 132 145 144 

30 F 47 104 116 116 119 127 142 151 145 153 154 155 154 154 

30 F 48 109 113 134 128 125 127 132 135 140 138 141 138 142 
B-1 

30 F 49 109 136 131 137 138 142 138 142 143 144 145 147 141 

30 F 50 100 112 119 119 135 135 133 135 135 136 130 140 140 

30 F 51 105 113 120 132 130 130 130 126 102 131 132 133 130 

30 F 52 107 134 133 129 139 136 143 144 145 147 144 131 150 

60 F 33 104 117 126 126 132 131 134 136 128 130 140 138 145 

60 F 34 119 114 127 129 128 135 124 137 132 134 138 139 142 

60 F 35 93 98 102 111 111 110 117 118 124 126 126 123 127 

60 F 36 101 107 116 115 133 134 133 137 122 134 137 127 139 

60 F 37 114 119 128 142 142 141 137 133 144 145 146 147 153 

60 F 38 99 103 125 131 132 137 133 131 142 137 112 132 125 

60 F 39 98 109 123 121 128 128 120 128 129 126 131 133 133 

60 F 40 109 107 110 117 115 127 133 139 142 140 138 142 144 

60 F 41 107 116 137 140 145 142 149 144 156 149 157 153 144 

60 F 42 102 132 137 132 142 140 134 138 150 145 149 145 151 

60 F 43 104 114 130 135 134 142 141 149 147 142 144 140 147 

60 F 44 101 115 130 123 121 136 140 140 145 145 145 143 145 

60 F 45 102 108 122 118 120 123 129 128 134 136 133 138 136 

60 F 46 102 124 126 126 140 142 134 144 147 147 142 147 

60 F 47 117 139 135 141 132 143 142 146 149 135 131 154 150 

60 F 48 103 116 141 138 140 137 137 137 145 146 147 155 

60 F 49 87 99 106 117 120 122 102 103 106 

60 F 50 99 120 122 121 127 126 118 131 112 99 121 134 138 

60 F 51 98 108 

60 F 52 114 138 138 141 137 144 142 147 150 150 147 153 151 

125 F 33 105 113 130 129 131 133 133 136 127 137 135 143 

125 F 34 112 136 136 136 130 140 142 144 148 145 148 150 150 

125 F 35 104 110 132 131 131 135 137 142 136 145 142 147 148 

125 F 36 101 123 120 123 125 121 132 135 134 136 136 136 136 

125 F 37 100 108 107 105 112 130 120 120 123 133 132 125 137 

125 F 38 116 113 131 133 142 137 148 139 142 155 146 159 

125 F 39 95 101 124 120 126 119 128 130 116 123 122 122 119 

125 F 40 106 129 133 140 136 141 143 148 138 129 146 139 140 

125 F 41 110 116 114 131 134 133 141 131 143 139 145 139 142 

125 F 42 94 105 108 123 126 129 129 131 135 133 135 137 131 

125 F 43 96 102 122 131 136 133 134 136 141 144 140 142 142 

125 F 44 104 123 130 127 132 132 140 132 142 142 146 136 143 

125 F 45 101 112 122 137 135 138 132 143 142 145 138 142 

125 F 46 108 110 127 123 128 130 126 130 131 133 133 132 133 

125 F 47 105 121 125 132 132 129 122 133 112 131 135 137 140 

125 F 48 109 137 138 139 137 136 138 139 140 148 148 146 146 

125 F 49 114 137 146 143 134 114 146 139 139 144 145 147 149 

125 F 50 97 111 131 125 133 128 142 137 133 128 137 137 142 

125 F 51 105 110 123 141 140 138 143 144 124 149 149 141 151 

125 F 52 103 115 135 128 135 136 136 139 141 138 140 139 

250 F 33 114 131 128 138 138 131 139 143 143 143 151 

250 F 34 107 127 132 143 141 146 143 152 150 153 151 

250 F 35 108 128 120 134 139 138 140 133 141 140 143 146 150 

250 F 36 112 130 129 134 136 132 133 250 116 143 146 
B-1 

250 F 37 98 110 131 121 131 133 132 131 127 133 134 

250 F 38 103 111 117 129 129 131 124 131 125 125 140 

250 F 39 97 104 117 127 128 129 124 131 133 127 135 

250 F 40 107 115 117 123 125 127 124 129 126 130 132 

250 F 41 108 117 125 126 132 119 113 132 132 139 140 143 141 

250 F 42 106 116 118 124 127 127 134 137 130 135 135 

250 F 43 101 108 120 129 131 132 137 134 138 109 120 123 134 

250 F 44 103 114 134 139 141 139 135 148 146 146 149 

250 F 45 122 120 133 153 158 153 150 152 160 160 155 163 163 

250 F 46 104 124 126 131 131 133 136 141 138 143 142 133 145 

250 F 47 97 118 128 124 131 124 135 133 114 132 123 163 130 

250 F 48 102 123 126 126 127 121 136 127 136 132 138 132 141 

250 F 49 103 114 122 130 135 132 139 135 137 142 143 145 145 

250 F 50 96 104 107 108 118 129 128 122 121 126 130 131 133 

250 F 51 108 120 134 136 142 142 146 150 106 154 158 

250 F 52 105 117 143 130 133 127 138 141 138 141 142 145 146 

0 M 153 88 100 101 107 104 106 108 110 108 114 114 116 120 

0 M 154 90 95 93 98 95 94 92 96 98 100 99 96 101 

0 M 155 87 97 103 103 104 105 108 106 107 107 107 108 106 

0 M 156 99 104 104 107 107 108 103 107 105 105 106 108 108 

0 M 157 104 107 109 110 112 113 110 113 116 114 114 115 115 

0 M 158 408 105 102 105 103 106 109 103 104 112 115 117 122 

0 M 159 95 104 102 104 107 107 107 109 110 111 111 112 109 

0 M 160 100 102 104 109 105 112 107 108 110 111 111 113 133 

0 M 161 105 116 117 117 119 118 108 112 113 116 114 116 121 

0 M 162 100 103 106 106 108 109 108 107 108 105 113 112 109 

0 M 163 95 98 103 103 102 107 106 109 107 107 109 107 107 

0 M 164 93 98 103 105 106 108 105 104 108 108 110 109 114 

0 M 165 98 102 102 104 107 106 107 108 110 109 111 111 112 

0 M 166 94 100 95 101 103 105 105 105 109 111 110 110 114 

0 M 167 104 106 109 108 109 109 111 109 108 107 109 110 111 

0 M 168 98 102 105 108 108 111 106 104 104 106 107 110 108 

0 M 169 101 107 110 112 111 114 113 122 124 126 120 129 134 

0 M 170 91 106 110 112 113 114 110 110 111 113 113 115 115 

0 M 171 104 106 109 112 112 113 114 117 117 119 117 118 120 

0 M 172 109 114 116 120 121 124 123 125 128 129 127 130 129 

30 M 53 114 119 124 110 115 121 122 129 131 133 137 140 141 

30 M 54 100 109 105 110 112 114 108 110 111 110 112 114 109 

30 M 55 104 110 112 112 110 116 114 118 117 118 118 118 119 

30 M 56 93 97 100 101 100 104 102 107 109 108 106 106 107 

30 M 57 101 108 108 108 107 108 105 107 109 109 111 111 109 

30 M 58 96 101 104 105 106 109 

30 M 59 100 105 113 103 105 110 107 114 116 113 97 110 121 

30 M 60 101 105 108 108 107 108 105 109 112 111 111 111 111 

30 M 61 101 105 104 105 106 111 107 111 108 106 109 110 110 

30 M 62 99 108 107 110 113 114 108 112 114 115 115 113 115 

30 M 63 105 105 101 102 103 104 106 110 107 108 112 115 115 

30 M 64 93 101 105 107 107 110 106 108 111 112 111 112 112 
B-1 

30 M 65 92 98 101 105 105 108 104 106 104 105 106 107 110 

30 M 66 103 108 108 109 110 112 112 115 112 112 113 115 116 

30 M 67 97 103 105 108 107 109 103 103 104 106 100 104 107 

30 M 68 108 118 122 125 129 129 127 130 132 131 131 130 132 

30 M 69 88 90 93 93 95 97 99 100 102 101 104 106 107 

30 M 70 94 98 100 99 100 99 96 100 101 102 102 106 

30 M 71 99 101 101 102 104 105 104 105 110 111 110 112 113 

30 M 72 102 107 107 112 115 116 115 121 120 118 121 122 121 

60 M 53 102 107 110 110 112 112 110 115 117 119 117 119 119 

60 M 54 102 107 106 110 112 111 110 110 114 115 118 117 

60 M 55 92 99 102 103 105 105 101 104 109 109 109 108 111 

60 M 56 96 100 103 101 103 107 104 106 108 108 102 104 110 

60 M 57 105 111 114 115 116 116 111 113 114 111 114 117 117 

60 M 58 97 103 103 106 109 110 107 109 111 113 113 115 117 

60 M 59 108 110 116 118 119 118 116 116 119 119 122 123 124 

60 M 60 96 98 99 101 102 104 109 109 113 113 114 111 115 

60 M 61 96 103 106 110 112 108 108 108 112 112 112 113 114 

60 M 62 96 102 104 108 111 110 109 109 112 112 114 111 115 

60 M 63 102 112 117 119 121 118 108 100 104 101 104 106 105 

60 M 64 99 104 103 101 103 106 104 106 112 110 111 109 112 

60 M 65 98 101 103 106 106 109 106 110 105 108 110 110 

60 M 66 100 101 105 108 111 113 111 112 114 116 116 112 114 

60 M 67 96 102 103 105 107 109 107 113 117 116 119 117 122 

60 M 68 105 109 109 110 109 112 111 113 117 112 116 120 119 

60 M 69 101 104 104 105 109 107 108 109 109 106 140 112 114 

60 M 70 98 97 101 105 109 110 106 102 113 110 112 110 115 

60 M 71 107 114 109 109 111 114 112 110 115 113 114 112 117 

60 M 72 87 93 95 96 97 99 99 100 99 101 101 102 102 

125 M 53 98 104 101 104 105 106 101 106 109 107 109 110 112 

125 M 54 103 106 109 110 113 114 112 111 115 117 119 119 

125 M 55 108 113 113 114 114 115 113 117 113 118 116 119 120 

125 M 56 107 114 119 117 114 115 109 110 112 112 115 115 115 

125 M 57 93 98 99 101 97 100 98 98 107 109 109 108 109 

125 M 58 106 108 101 108 107 111 112 117 120 120 119 121 122 

125 M 59 101 93 97 99 100 99 96 98 100 102 99 101 101 

125 M 60 101 101 99 98 99 110 100 105 101 103 103 106 108 

125 M 61 96 96 99 99 101 103 100 101 103 104 105 105 104 

125 M 62 97 99 106 100 108 109 108 113 116 119 121 122 120 

125 M 63 94 99 101 102 100 98 103 104 102 106 108 108 109 

125 M 64 98 104 104 107 110 113 106 108 111 110 115 111 109 

125 M 65 90 90 89 87 88 89 90 93 93 94 94 93 97 

125 M 66 101 99 103 103 103 105 104 105 106 107 107 108 107 

125 M 67 105 109 111 112 112 112 111 113 17 115 115 109 118 

125 M 68 94 109 97 100 101 103 100 98 98 112 102 104 104 

125 M 69 96 101 103 104 105 107 107 106 110 111 111 113 113 

125 M 70 103 103 100 99 104 105 100 102 101 103 130 104 104 

125 M 71 105 104 105 106 107 106 107 107 107 110 112 114 114 

125 M 72 104 111 108 109 111 111 110 109 109 112 113 115 116 
B-1 

250 M 53 100 97 100 102 106 109 112 116 109 114 115 119 119 

250 M 54 94 99 102 103 107 107 104 108 108 109 108 110 109 

250 M 55 92 96 99 99 100 100 98 95 101 104 104 103 107 

250 M 56 90 96 97 100 102 104 101 103 104 105 105 107 108 

250 M 57 98 101 102 104 107 106 100 105 105 104 

250 M 58 108 117 121 123 124 125 122 125 125 126 131 

250 M 59 103 106 107 112 112 114 114 110 111 119 122 

250 M 60 98 101 102 104 105 108 106 109 109 108 111 

250 M 61 95 99 101 100 103 103 106 107 107 111 

250 M 62 101 104 104 105 107 106 102 104 105 105 108 

250 M 63 106 108 110 110 113 111 110 114 111 111 113 117 118 

250 M 64 94 93 100 104 108 110 108 109 108 109 112 112 109 

250 M 65 91 102 95 105 100 106 103 106 106 107 107 107 110 

250 M 66 105 102 109 114 114 117 107 116 115 117 118 117 

250 M 67 102 106 110 112 112 115 110 114 116 114 119 118 119 

250 M 68 97 101 101 103 103 105 110 107 106 108 109 111 111 

250 M 69 104 111 111 113 113 112 109 111 113 115 114 112 118 

250 M 70 90 91 96 97 95 98 101 105 103 104 106 105 109 

250 M 71 92 100 100 95 103 100 105 105 104 108 105 109 110 

250 M 72 99 107 102 102 102 104 103 107 107 107 109 112 117 

P generation cloaca measurements and foam

      week 6 ‐ 4/20/09     week 7 ‐ 4/27/09    

DOSE ID cg area foam cg area Foam

0 153 72     150 Y

0 154 36     100    

0 155 36     108 Y

0 156 56     130 Y

0 157 154 Y 234 Y

0 158 154 Y 195 Y

0 159 25     117 Y

0 160 144 Y 224 Y

0 161 99     182 Y

0 162 99     154 Y

0 163 182 Y 208 Y

0 164 80 Y 154 Y

0 165 182 Y 195 Y

0 166 81     96 Y

0 167 195 Y 195 Y

0 168 154 Y 195 Y

0 169 154 Y 224 Y

0 170 49     110 Y

0 171 195 Y 208 Y

0 172 120 Y 108 Y

30 53 90     108 Y

30 54 25     24    

30 55 25     81    

30 56 117 Y 221 Y
B-1 

30 57 25     25    

30 58 72     90    

30 59 25     25    

30 60 100     180Y

30 61 25     25Y

30 62 25     64    

30 63 72     99Y

30 64 49     144Y

30 65 121Y 168Y

30 66 120Y 154Y

30 67 54     108Y

30 68 90     130Y

30 69 81     130Y

30 70 140Y 240Y

30 71 180Y 224Y

30 72 72Y 132Y

60 53 132     195Y

60 54 25     80    

60 55 72Y 126Y

60 56 25     36    

60 57 72     180Y

60 58 60     120Y

60 59 90     117Y

60 60 144Y 182Y

60 61 72     165Y

60 62 25     25    

60 63 25     25    

60 64 25     25    

60 65 64     121    

60 66 25     25    

60 67 120Y 192Y

60 68 120Y 180Y

60 69 90Y 135Y

60 70 132Y 121Y

60 71 56     208Y

60 72 25     25    

125 53 168Y 221Y

125 54 90     182Y

125 55 90Y 182Y

125 56 80     121Y

125 57 108Y 182Y

125 58 154Y 210Y

125 59 99Y 117Y

125 60 90     192Y

125 61 88     154Y

125 62 25     25    

125 63 121Y 208Y

125 64 72     165Y
B-1 

125 65 180 Y 204 Y

125 66 90 Y 154 Y

125 67 25     120    

125 68 63     168 Y

125 69 72     154 Y

125 70 120 Y 192 Y

125 71 120 Y 195 Y

125 72 99 Y 154 Y

250 62 120 Y 182 Y

250 55 70     132 Y

250 56 99     168 Y

250 57 180 Y 208 Y

250 58 25     36    

250 59 120 Y 224 Y

250 60 180 Y 221 Y

250 61 25     99    

250 66 63     121 Y

250 68 25     25    

250 69 110 Y 165 Y

250 70 64     180 Y

250 71 92     165 Y

250 72 25     25    

250 65 25     25    

250 67 154 Y 120 Y

250 54 108     154 Y

250 53 25     120 Y

250 63 81     210 Y

250 64 90     154 Y

P generation egg strength 

DOSE ID Thickness Thickness 

Avg. 

thickness Strength 

0 135 1.57 1.05 1.31 1.374 

0 137 1.36 0.78 1.07 1.302 

0 138 0.77 0.92 0.845 0.963 

0 140 1.49 0.98 1.235 1.041 

0 141 0.44 0.68 0.56 1.194 

0 142 1.32 1.16 1.24 0.874 

0 143 1.59 1.94 1.765 1.013 

0 144 1.02 1.42 1.22 0.855 

0 147 1.21 1.43 1.32 1.56 

0 148 0.91 1.4 1.155 0.354 

0 149 0.89 1.07 0.98 0.982 

0 150 1.28 0.104 0.692 1.324 

0 135 0.76 0.37 0.565 0.853 

0 138 0.96 0.42 0.69 0.506 

0 140 0.75 0.59 0.67 0.879 

0 142 0.79 0.43 0.61 0.819 

0 143 0.47 0.37 0.42 0.454 
B-1 

0 144 0.98 0.53 0.755 1.107 

0 147 1.43 1.06 1.245 1.122 

0 149 0.39 0.57 0.48 0.57 

0 150 0.63 0.85 0.74 0.44 

0 141 1.16 0.73 0.875 

30 33 0.68 1.07 0.875 0.864 

30 34 1.23 1.1 1.165 1.248 

30 37 1.16 1.19 1.175 0.367 

30 38 1.22 1.35 1.285 1.365 

30 41 2.03 1.59 1.81 1.94 

30 43 1.37 1.15 1.26 1.302 

30 49 1.56 1.22 1.39 1.192 

30 51 1.17 1.18 1.175 0.325 

30 33 0.85 0.91 0.88 1.399 

30 44 0.58 0.96 0.77 0.908 

30 34 0.38 0.38 0.38 1.36 

30 35 0.46 0.77 0.615 1.246 

30 36 1.42 1.66 1.54 1.475 

30 38 0.47 0.38 0.425 0.778 

30 40 0.78 0.95 0.865 1.361 

30 41 0.37 0.41 0.39 1.446 

30 42 1.16 1.01 1.085 0.433 

30 43 0.47 0.5 0.485 0.85 

30 44 0.92 0.67 0.795 1.131 

30 45 0.84 0.63 0.735 0.691 

30 46 0.7 0.48 0.59 1.015 

30 47 0.38 0.51 0.445 1.032 

30 48 0.42 0.45 0.435 0.709 

30 49 0.73 0.81 0.77 1.037 

30 50 1.17 0.75 0.96 1.109 

30 51 0.36 0.38 0.37 1.005 

30 52 0.5 0.32 0.41 1.031 

30 35 1.22 0.84 1.41 

30 36 0.7 1.39 1.637 

30 37 0.65 0.55 0.725 

30 39 1.08 0.84 0.537 

30 42 0.84 0.7 0.509 

30 46 0.89 0.71 0.845 

30 47 2.54 0.45 1.245 

30 48 1.01 1.02 0.955 

30 50 1.11 0.96 1.22 

30 52 1.39 0.8 1.227 

60 33 2.95 2.87 2.91 1.435 

60 35 0.94 1.29 1.115 0.671 

60 37 1.36 1.42 1.39 1.295 

60 42 1.6 1.5 1.55 1.256 

60 43 4.46 3.6 4.03 1.484 

60 46 1.48 0.76 1.12 1.198 
B-1 

60 52 0.3 0.54 0.42 0.938 

60 50 1.51 0.95 1.23 0.687 

60 33 0.47 0.43 0.45 1.119 

60 36 0.3 0.35 0.325 0.823 

60 37 0.77 0.76 0.765 0.813 

60 38 0.42 0.32 0.37 1.193 

60 39 0.34 0.29 0.315 1.179 

60 41 0.51 0.42 0.465 0.974 

60 42 1.05 0.99 1.02 1.627 

60 43 0.84 0.72 0.78 1.292 

60 44 0.36 0.27 0.315 1.62 

60 45 0.37 0.38 0.375 0.9 

60 47 1.2 0.78 0.99 0.931 

60 48 1.24 0.98 1.11 0.957 

60 50 0.79 0.46 0.625 1.101 

60 35 0.91 0.52 0.803 

60 36 1.1 0.91 0.605 

60 40 0.67 0.76 0.706 

60 41 0.42 0.46 1.104 

60 44 1.42 1.01 1.179 

60 45 0.88 0.84 0.888 

60 48 1.12 0.79 0.729 

60 52 1.12 0.61 0.969 

125 33 0.51 0.52 0.515 1.016 

125 35 1.36 0.67 1.015 1.026 

125 36 1.01 0.42 0.715 1.319 

125 39 1.29 1.05 1.17 1.216 

125 40 0.45 0.37 0.41 1.315 

125 41 0.71 1.17 0.94 1.27 

125 42 1.07 1.4 1.235 0.383 

125 44 0.39 0.95 0.67 1.026 

125 46 0.52 0.86 0.69 1.54 

125 47 0.63 0.96 0.795 1.309 

125 48 0.4 1.45 0.925 1.388 

125 49 0.69 0.8 0.745 1.53 

125 50 0.94 0.67 0.805 1.239 

125 51 1.25 1.08 1.165 1.644 

125 52 1.44 1.24 1.34 1.57 

125 33 0.8 1.1 0.95 1.148 

125 39 0.66 1.05 0.855 1.337 

125 40 1.31 1.53 1.42 0.709 

125 41 0.86 0.47 0.665 1.604 

125 43 1.3 1.67 1.485 1.314 

125 45 1.15 1.06 1.105 1.261 

125 52 0.42 0.4 0.41 1.3 

125 34 0.42 0.33 0.375 1.051 

125 35 0.71 0.59 0.65 0.341 

125 36 0.91 0.44 0.675 1.258 
B-1 

125 37 0.61 1.03 0.82 1.375 

125 38 1.32 1.16 1.24 1.455 

125 39 0.61 0.55 0.58 1.17 

125 42 0.43 0.39 0.41 1.3 

125 43 0.87 0.67 0.77 0.823 

125 44 1.14 0.81 0.975 1.11 

125 45 0.48 0.28 0.38 1.233 

125 46 0.66 1.03 0.845 1.41 

125 48 0.68 0.39 0.535 1.143 

125 49 0.56 0.86 0.71 1.256 

125 50 0.88 1.41 1.145 0.914 

125 51 0.39 0.37 0.38 1.2 

125 34 1.09 1.47 0.824 

125 35 0.73 0.43 0.906 

125 38 1.68 1.22 1.275 

250 33 1.88 0.97 1.425 1.418 

250 34 1.77 1.01 1.39 1.566 

250 36 0.34 1.05 0.695 1.198 

250 37 0.75 0.45 0.6 0.773 

250 38 0.93 1.25 1.09 1.009 

250 43 0.42 0.95 0.685 1.308 

250 44 1.07 0.53 0.8 1.012 

250 45 0.78 0.79 0.785 1.6 

250 46 1.04 0.67 0.855 1.487 

250 47 1.71 1.77 1.74 

250 48 1.45 0.87 1.16 1.13 

250 49 0.56 0.82 0.69 1.113 

250 52 1.11 1.17 1.14 1.356 

250 33 1.25 1.81 1.53 0.824 

250 35 0.36 1 0.68 0.802 

250 39 0.63 0.61 0.62 1.23 

250 41 1.07 0.71 0.89 1.739 

250 43 1.55 1.27 1.41 0.919 

250 44 1.25 1.16 1.205 0.837 

250 46 0.77 0.67 0.72 1.475 

250 52 0.63 0.69 0.66 1.136 

250 35 0.36 0.41 0.385 0.848 

250 37 0.38 0.37 0.375 0.514 

250 45 1 0.92 0.96 1.49 

250 49 0.57 0.46 0.515 0.889 

250 50 0.45 0.31 0.38 0.688 

250 51 0.63 0.51 0.57 0.471 

250 52 0.67 0.45 0.56 1.457 

250 38 0.83 0.8 0.571 

250 42 0.89 0.56 1.21 

P generation yolk content 

 ug/g yolk 
B-1 

DOSE ID total endosulfans I II 

0 140A 0 0.0128 0.0017 

0 150A 0 0.0113 0.0014 

0 141A 0.0007 0.0123 0.0023 

30 36A 0.0813 0.0227 0.08 

30 49A 0.1528 0.0285 0.1449 

30 50A 0.1623 0.0371 0.1505 

30 42A 0.0847 0.0177 0.0865 

60 52A 0.3268 0.0465 0.316 

60 36A 0.3929 0.0695 0.3615 

60 47A 0.4758 0.0741 0.448 

120 50A 0.6566 0.0856 0.6392 

120 42A 0.5114 0.0638 0.5012 

120 52A 0.5573 0.0947 0.5143 

120 45A 1.2275 0.1587 1.1989 

120 48A 0.8998 0.1147 0.8839 

250 36A 1.8063 0.2385 1.7664 

250 52A 1.4989 0.1734 1.4914 

250 42A 1.3745 0.2301 1.2737 

250 37A 2.3351 0.3186 2.3012 

250 50A 1.2628 0.1304 1.3126 

P generation sperm measurements 

DOSE ID sperm/gram % motile % progressive 

0 164 7735.85 97.8 16.5 

0 163 8797.65 94.5 17.5 

0 156 8223.95 95.5 18.5 

0 157 5467 87 11.5 

0 155 7047.45 89.5 12.5 

0 170 6363.6 95 29.5 

0 169 6361.6 89.5 18 

0 171 7945.37 93 11.33 

0 154 5994.45 97 21 

0 153 5240.5 96.5 19.5 

30 56 3236.47 87 20.67 

30 66 8328.07 95 24.33 

30 71 5467 84.5 13.5 

30 67 1744.23 22 2 

30 64 5646.4 92 17 

30 59 4725.9 94.3 20.7 

30 63 4659.6 92.3 21.7 

30 65 4325 93.7 16.3 

30 68 9375.9 97.5 19.5 

60 65 4943.7 93.5 22 

60 64 6478.4 92 19.5 

60 68 1827.93 93.67 18 

60 67 4045.6 84.33 11 

60 63 6666.43 89 13 
B-1 

60 66 9476.13 134.5 21 

60 72 8245.03 96.67 17 

60 70 4214.53 75 14 

60 71 6421.9 97 20.67 

60 69 3977.9 87.67 11.33 

125 69 5904.35 95 15 

125 67 5259.85 98.5 33 

125 65 4191.35 91 16.5 

125 70 6747.8 94.5 19.5 

125 71 4346.25 86.5 17 

125 63 5432.85 96 19 

125 66 2502.65 67.5 10.5 

125 72 1858.8 85.5 26 

125 68 1664.4 67 6.5 

250 69 5600.65 97 26.5 

250 72 4764.1 89 14.5 

250 71 6463.95 95.5 18 

250 61 6542.2 96 20 

250 60 3549.95 96 18.5 

250 65 2763.35 70.5 9 

250 62 6411.3 92 18.5 

250 56 5426 85.5 13 

250 70 4668.8 74 12 
B-1 
P generation feed consumption 

 4/21 

week 1 consumption 

4/21 4/28 

week 2 consumption 

4/28 5/5 

week 3 consumption 

5/5 5/12 

week 4 consumption 

5/12 5/19 

week 5 consum

DOSE SEX ID initial final Initial final initial final initial final initial final 0 F 144 419 315 104 430 312 118 455 332 123 444 335 109 449 320 129 0 F 142 415 305 110 427 296 131 425 312 113 447 331 116 439 258 181 0 F 140 419 282 137 413 259 154 454 284 170 459 266 193 454 259 195 0 F 139 424 324 100 425 320 105 454 341 113 448 338 110 454 342 112 0 F 141 382 271 111 422 297 125 450 333 117 446 258 188 443 255 188 0 F 143 426 310 116 418 312 106 460 311 149 431 237 194 465 267 198 0 F 150 418 320 98 423 315 108 457 327 130 440 315 125 435 247 188 0 F 149 406 285 121 422 327 95 462 310 152 440 257 183 450 450 0 F 148 402 304 98 419 309 110 456 331 125 436 294 142 450 289 161 0 F 147 402 296 106 416 285 131 454 261 193 439 269 170 483 282 201 0 F 146 409 278 131 426 312 114 461 355 106 453 370 83 459 386 73 0 F 145 395 242 153 425 307 118 460 307 153 433 245 188 433 243 190 0 F 137 406 303 103 415 315 100 458 322 136 429 253 176 469 278 191 0 F 135 388 268 120 416 270 146 436 246 190 443 246 197 441 236 205 0 F 133 410 308 102 431 303 128 439 322 117 438 260 178 444 328 116 0 F 134 396 259 137 424 321 103 446 0 F 136 404 337 67 427 277 150 443 286 157 427 267 160 436 269 167 0 F 138 421 309 112 425 327 98 458 321 137 432 272 160 445 273 172 0 F 151 416 304 112 418 337 81 445 330 115 441 330 111 446 446 0 F 152 426 302 124 424 348 76 457 30 F 49 396 288 108 437 307 130 445 279 166 435 268 167 476 269 207 30 F 50 393 280 113 449 293 156 460 315 145 439 306 133 442 295 147 30 F 51 402 261 141 438 310 128 440 334 106 478 287 191 467 264 203 30 F 52 430 299 131 433 273 160 438 253 185 439 260 179 464 276 188 30 F 33 402 281 121 441 307 134 441 282 159 449 296 153 442 290 152 30 F 34 404 282 122 433 306 127 443 303 140 430 272 158 425 281 144 30 F 35 435 299 136 437 315 122 443 344 99 436 353 83 450 375 75 30 F 36 403 267 136 437 308 129 457 271 186 448 315 133 450 305 145 30 F 37 418 273 145 435 328 107 451 356 95 432 366 66 444 357 87 30 F 38 409 247 162 444 282 162 456 249 207 433 251 182 425 233 192 30 F 39 414 261 153 430 266 164 453 254 199 432 247 185 461 239 222 30 F 40 406 272 134 436 278 158 463 337 126 431 249 182 450 269 181 30 F 41 422 291 131 437 295 142 448 258 190 433 254 179 429 236 193 30 F 42 404 272 132 433 283 150 454 266 188 445 269 176 427 248 179 30 F 43 415 286 129 437 310 127 452 320 132 431 264 167 447 236 211 30 F 44 427 309 118 442 311 131 455 295 160 463 286 177 421 242 179 30 F 45 401 262 139 437 331 106 455 341 114 441 350 91 437 349 88 30 F 46 413 267 146 439 325 114 452 335 117 432 349 83 443 341 102 30 F 47 421 316 105 440 305 135 455 355 100 440 358 82 446 347 99 30 F 48 411 274 137 439 326 113 447 339 108 435 282 153 415 273 142 60 F 33 404 281 123 436 319 117 444 441 303 138 60 F 34 391 238 153 439 257 182 443 248 195 430 286 144 437 267 170 60 F 35 396 302 94 438 324 114 442 358 84 440 354 86 448 327 121 60 F 36 396 290 106 430 320 110 436 312 124 439 321 118 440 329 111 60 F 37 407 260 147 433 311 122 443 309 134 438 265 173 474 263 211 60 F 38 390 281 109 429 330 99 444 312 132 429 289 140 419 278 141 
B-1 

60 F 39 401 292 109 437 314 123 440 268 172 433 273 160 428 274 154 60 F 40 407 284 123 427 346 81 452 268 184 441 344 97 450 350 100 60 F 47 414 266 148 440 303 137 445 273 172 426 254 172 408 261 147 60 F 48 445 324 121 434 306 128 445 308 137 428 258 170 417 262 155 60 F 49 406 322 84 436 340 96 447 338 109 437 314 123 423 317 106 60 F 50 429 307 122 435 321 114 443 292 151 436 288 148 425 259 166 60 F 51 420 321 99 435 341 94 60 F 52 414 297 117 435 276 159 439 254 185 437 254 183 418 231 187 60 F 41 393 269 124 428 302 126 452 269 183 431 267 164 419 236 183 60 F 42 423 316 107 434 290 144 441 251 190 448 267 181 418 238 180 60 F 43 395 280 115 438 323 115 450 284 166 440 265 175 413 239 174 60 F 44 405 294 111 435 322 113 440 283 157 435 266 169 430 279 151 60 F 45 411 301 110 436 328 108 447 321 126 422 285 137 428 325 103 60 F 46 426 296 130 437 282 155 430 254 176 470 238 232 481 247 234 125 F 38 417 266 151 431 323 108 434 265 169 488 269 219 473 235 238 125 F 39 431 333 98 431 322 109 430 306 124 434 282 152 439 295 144 125 F 37 421 283 138 427 207 220 423 292 131 446 278 168 430 295 135 125 F 36 411 284 127 430 241 189 430 251 179 481 262 219 475 247 228 125 F 35 391 282 109 430 302 128 434 259 175 442 243 199 432 236 196 125 F 34 407 285 122 435 286 149 439 274 165 437 243 194 466 272 194 125 F 33 410 286 124 429 304 125 425 307 118 404 257 147 439 279 160 125 F 44 407 290 117 437 276 161 431 298 133 474 251 223 516 249 267 125 F 43 410 309 101 437 325 112 438 293 145 487 267 220 419 125 F 42 411 311 100 430 313 117 444 342 102 442 309 133 430 296 134 125 F 41 405 247 158 423 278 145 443 350 93 431 266 165 467 252 215 125 F 40 410 296 114 435 281 154 420 252 168 431 248 183 431 236 195 125 F 45 426 312 114 435 276 159 436 273 163 435 255 180 426 250 176 125 F 46 417 289 128 430 324 106 446 348 98 434 279 155 417 232 185 125 F 47 412 304 108 431 309 122 454 305 149 444 265 179 422 247 175 125 F 48 425 294 131 441 289 152 437 253 184 480 248 232 420 234 186 125 F 49 407 270 137 432 295 137 450 285 165 436 243 193 427 241 186 125 F 50 411 301 110 437 294 143 436 286 150 440 244 196 460 251 209 125 F 52 410 231 179 434 264 170 437 254 183 433 247 186 429 241 188 125 F 51 401 305 96 432 316 116 439 300 139 442 245 197 467 245 222 250 F 43 393 272 121 429 312 117 424 264 160 471 439 263 176 250 F 44 398 283 115 428 280 148 429 281 148 434 251 183 433 269 164 250 F 45 402 273 129 433 316 117 429 333 96 440 292 148 432 294 138 250 F 46 410 262 148 436 239 197 432 232 200 439 253 186 433 239 194 250 F 47 405 303 102 438 309 129 442 295 147 436 268 168 440 250 F 48 401 246 155 430 281 149 430 302 128 442 253 189 438 259 179 250 F 49 405 268 137 428 241 187 436 243 193 474 244 230 473 250 223 250 F 50 402 291 111 445 330 115 447 356 91 442 355 87 448 360 88 250 F 51 401 274 127 424 283 141 421 240 181 472 261 211 437 234 203 250 F 52 393 252 141 435 303 132 433 239 194 472 244 228 440 240 200 250 F 36 403 278 125 419 248 171 423 254 169 437 243 194 455 249 206 250 F 35 408 290 118 421 289 132 427 302 125 448 274 174 414 242 172 250 F 34 398 270 128 423 257 166 420 258 162 479 243 236 457 231 226 250 F 33 397 278 119 427 267 160 431 278 153 446 267 179 441 261 180 250 F 37 399 271 128 430 300 130 424 317 107 434 287 147 442 303 139 250 F 38 407 295 112 446 300 146 421 295 126 429 285 144 424 246 178 250 F 39 401 256 145 435 245 190 427 235 192 441 267 174 434 246 188 250 F 40 404 295 109 431 302 129 431 284 147 446 300 146 430 304 126 
B-1 

250 F 41 398 271 127 429 307 122 429 343 86 448 305 143 436 267 169 250 F 42 408 298 110 438 304 134 426 275 151 433 246 187 429 245 184 0 M 158 419 297 122 431 351 80 462 351 111 437 329 108 446 335 111 0 M 157 417 290 127 431 281 150 451 272 179 438 281 157 434 281 153 0 M 156 412 307 105 432 305 127 456 263 193 437 286 151 442 319 123 0 M 155 424 343 81 424 313 111 449 322 127 438 312 126 443 323 120 0 M 154 426 323 103 416 310 106 457 353 104 442 331 111 440 330 110 0 M 153 419 335 84 430 304 126 450 336 114 434 312 122 440 329 111 0 M 164 427 328 99 423 311 112 463 344 119 436 324 112 445 339 106 0 M 163 431 337 94 427 329 98 467 365 102 435 339 96 441 336 105 0 M 162 407 279 128 424 293 131 452 308 144 444 311 133 435 300 135 0 M 161 420 260 160 435 234 201 455 261 194 441 295 146 442 275 167 0 M 160 414 313 101 432 322 110 454 342 112 430 314 116 442 333 109 0 M 159 410 301 109 423 294 129 454 330 124 434 305 129 445 304 141 0 M 165 409 302 107 418 325 93 456 351 105 432 337 95 453 339 114 0 M 166 428 316 112 428 312 116 434 328 106 434 437 328 109 0 M 167 399 279 120 428 286 142 456 275 181 430 304 126 450 334 116 0 M 168 412 315 97 437 340 97 459 351 108 447 341 106 432 328 104 0 M 169 426 320 106 437 303 134 449 329 120 430 325 105 437 325 112 0 M 170 399 314 85 437 306 131 446 295 151 438 269 169 429 273 156 0 M 171 414 315 99 435 319 116 458 332 126 436 323 113 452 344 108 0 M 172 421 312 109 440 326 114 440 326 114 427 309 118 452 335 117 30 M 63 395 267 128 437 264 173 418 309 109 432 279 153 429 300 129 30 M 64 408 310 98 444 332 112 455 341 114 437 332 105 443 350 93 30 M 65 415 314 101 434 326 108 447 326 121 435 320 115 444 346 98 30 M 66 408 290 118 441 335 106 453 341 112 444 333 111 458 349 109 30 M 67 405 282 123 442 332 110 448 351 97 430 344 86 439 365 74 30 M 68 424 312 112 438 303 135 450 308 142 432 300 132 413 299 114 30 M 72 409 275 134 440 311 129 444 291 153 438 306 132 456 305 151 30 M 71 417 309 108 444 335 109 452 305 147 434 330 104 457 307 150 30 M 70 409 303 106 445 343 102 446 329 117 426 351 75 459 366 93 30 M 69 401 308 93 442 345 97 446 331 115 402 324 78 449 442 7 30 M 53 417 305 112 433 317 116 450 315 135 435 351 84 450 339 111 30 M 54 405 307 98 440 337 103 452 362 90 437 330 107 453 352 101 30 M 55 422 309 113 442 313 129 446 317 129 433 314 119 448 337 111 30 M 56 410 304 106 439 335 104 449 329 120 441 341 100 431 335 96 30 M 57 420 311 109 439 310 129 441 328 113 436 329 107 432 307 125 30 M 58 420 311 109 444 326 118 447 318 129 438 314 124 436 313 123 30 M 59 418 301 117 437 327 110 446 312 134 440 362 78 447 357 90 30 M 60 411 309 102 441 321 120 448 322 126 432 320 112 440 324 116 30 M 61 413 323 90 451 349 102 452 354 98 433 337 96 450 30 M 62 406 293 113 439 306 133 440 302 138 434 305 129 432 294 138 60 M 54 418 314 104 439 315 124 441 305 136 431 292 139 443 302 141 60 M 53 410 293 117 433 312 121 448 311 137 430 309 121 440 330 110 60 M 72 419 331 88 430 336 94 440 339 101 430 324 106 447 338 109 60 M 71 411 265 146 430 310 120 432 351 81 431 353 78 429 321 108 60 M 70 408 294 114 436 344 92 455 321 134 453 313 140 441 322 119 60 M 69 411 303 108 434 333 101 427 321 106 431 310 121 419 315 104 60 M 68 415 295 120 434 314 120 432 330 102 434 326 108 441 319 122 60 M 67 415 304 111 433 311 122 435 313 122 445 323 122 443 333 110 60 M 55 415 291 124 434 314 120 438 322 116 436 329 107 444 275 169 60 M 56 414 309 105 437 337 100 449 334 115 429 339 90 450 352 98 
B-1 

60 M 57 403 284 119 436 319 117 442 312 130 435 324 111 445 346 99 60 M 58 398 288 110 429 312 117 452 335 117 432 319 113 435 329 106 60 M 59 408 291 117 440 325 115 454 301 153 442 307 135 435 320 115 60 M 60 400 296 104 440 321 119 447 310 137 431 290 141 438 294 144 60 M 61 410 310 100 436 323 113 444 318 126 438 326 112 449 337 112 60 M 62 416 310 106 432 331 101 448 333 115 441 343 98 445 352 93 60 M 63 417 289 128 427 289 138 450 330 120 434 337 97 441 362 79 60 M 64 432 303 129 432 306 126 437 313 124 432 307 125 435 292 143 60 M 65 410 299 111 427 319 108 452 319 133 433 315 118 441 324 117 60 M 66 406 297 109 430 334 96 447 326 121 433 297 136 438 315 123 125 M 58 411 288 123 436 328 108 445 359 86 438 322 116 435 327 108 125 M 57 408 308 100 444 319 125 448 448 432 336 96 125 M 56 415 285 130 438 306 132 441 441 444 354 90 125 M 55 414 294 120 434 319 115 451 451 444 343 101 125 M 54 409 301 108 444 335 109 430 430 432 318 114 125 M 53 408 299 109 439 335 104 436 436 443 345 98 125 M 59 410 280 130 434 370 64 452 311 141 440 303 137 436 325 111 125 M 60 408 294 114 430 320 110 434 317 117 452 331 121 448 331 117 125 M 61 433 329 104 447 332 115 440 320 120 445 320 125 439 322 117 125 M 62 409 299 110 439 318 121 434 309 125 435 336 99 429 313 116 125 M 63 400 300 100 439 318 121 438 307 131 454 328 126 435 336 99 125 M 64 420 326 94 438 325 113 430 330 100 437 322 115 431 322 109 125 M 69 406 303 103 445 306 139 432 293 139 441 273 168 426 266 160 125 M 70 401 271 130 434 358 76 436 345 91 443 339 104 432 324 108 125 M 71 399 269 130 424 296 128 436 436 423 324 99 125 M 72 400 270 130 435 302 133 432 339 93 440 330 110 443 321 122 125 M 68 406 298 108 443 345 98 439 321 118 437 305 132 427 308 119 125 M 66 405 299 106 435 337 98 428 331 97 435 341 94 437 338 99 125 M 67 395 287 108 435 314 121 422 299 123 442 330 112 438 339 99 125 M 65 401 300 101 426 337 89 420 332 88 440 339 101 442 331 111 250 M 57 405 310 95 439 337 102 445 322 123 431 323 108 443 331 112 250 M 56 399 292 107 438 317 121 422 276 146 438 308 130 450 305 145 250 M 55 398 293 105 441 340 101 445 326 119 439 324 115 435 325 110 250 M 62 403 290 113 427 313 114 422 300 122 442 303 139 433 309 124 250 M 58 403 297 106 436 310 126 420 284 136 432 302 130 447 313 134 250 M 59 439 306 133 433 290 143 422 290 132 441 305 136 442 328 114 250 M 60 397 298 99 426 325 101 429 316 113 440 315 125 437 312 125 250 M 61 395 281 114 428 309 119 429 293 136 439 309 130 442 306 136 250 M 66 409 300 109 442 339 103 434 317 117 437 319 118 434 330 104 250 M 68 409 294 115 445 335 110 431 301 130 432 306 126 442 322 120 250 M 71 426 319 107 438 322 116 434 326 108 439 336 103 450 346 104 250 M 72 411 298 113 438 303 135 440 353 87 446 352 94 443 339 104 250 M 70 403 307 96 444 355 89 443 340 103 437 341 96 432 349 83 250 M 69 409 297 112 438 307 131 441 298 143 440 302 138 439 312 127 250 M 65 392 298 94 440 327 113 423 341 82 444 344 100 434 344 90 250 M 67 406 305 101 442 337 105 439 332 107 434 435 324 111 250 M 54 404 304 100 440 327 113 421 307 114 437 316 121 441 325 116 250 M 53 403 296 107 424 343 81 404 339 65 434 328 106 434 333 101 250 M 63 400 282 118 428 314 114 421 299 122 441 434 302 132 250 M 64 409 321 88 444 353 91 431 338 93 447 340 107 433 336 97 

P generation feed consumption – paired 
B-1 

 5/29 6/2 6/5 6/9 6/12 6/16 6/19 6/22 6/26 6/30 7/3 7/7 7/10 Dose 

Male 

Female 

Initial 

Initial 

Final 

Initial 

Final 

Initial Final 

Initial 

Final 

Initial 

Final 

Initial 

Final 

Initial Final 

Initial 

Final 

Initial Final 

Initial 

Final 

Initial 

Final 

Initl 

0 136 414 317 416 350 421 315 425 337 426 327 435 354 420 321 432 347 425 332 430 351 429 333 422 344 0 143 405 257 414 347 417 284 411 326 406 311 422 310 420 270 420 312 384 260 414 332 416 252 434 355 0 153 417 335 436 381 425 355 433 376 430 361 431 386 412 335 417 352 402 337 438 390 409 245 438 384 0 154 139 410 272 434 300 416 251 397 269 407 258 423 309 425 278 421 307 405 258 420 267 417 266 403 276 0 155 141 405 236 418 270 422 255 402 259 416 256 427 260 428 241 420 292 403 263 422 281 418 253 417 306 0 156 135 413 231 420 271 420 239 397 244 416 237 415 252 428 241 406 245 421 230 422 260 396 270 431 336 0 157 142 421 418 270 399 246 410 311 425 240 414 270 404 258 416 307 417 242 419 268 423 254 417 288 0 158 144 418 254 409 291 413 266 399 299 424 284 412 302 418 288 416 346 415 337 436 367 425 351 422 356 0 164 145 415 268 427 301 417 241 395 246 427 245 428 307 423 244 425 270 412 254 415 291 405 246 429 260 0 163 146 424 311 433 339 401 278 422 308 423 271 425 314 433 272 432 329 409 274 410 290 408 255 414 286 0 162 140 413 255 422 280 408 236 401 241 418 247 410 278 416 251 404 264 411 243 403 235 415 254 433 261 0 161 148 412 234 419 271 404 252 404 233 414 229 410 229 424 232 427 231 410 242 419 263 397 233 422 292 0 160 149 411 269 424 297 417 264 400 273 410 254 409 294 423 258 409 306 409 255 416 276 409 264 413 284 0 159 150 420 249 421 276 424 258 401 257 421 239 409 253 423 242 431 252 406 251 410 267 413 254 419 272 0 165 137 421 315 431 334 435 284 406 286 413 253 429 307 448 280 413 290 418 260 418 268 405 255 422 290 0 166 412 347 426 377 436 368 420 367 420 361 434 390 441 375 426 385 415 363 428 385 416 368 438 391 0 167 133 415 252 415 305 425 238 407 241 421 243 424 326 430 257 420 315 413 259 410 237 402 248 407 258 0 168 147 413 241 409 268 417 264 403 252 413 242 425 228 429 256 429 268 422 414 249 410 244 423 248 0 169 412 333 418 349 434 344 411 339 418 344 430 369 439 350 420 363 413 356 433 361 411 342 418 350 0 170 421 342 414 365 425 344 417 359 416 277 425 379 436 346 419 366 421 342 423 374 413 328 436 362 0 171 426 358 425 375 432 365 431 377 417 358 429 383 437 371 413 366 419 366 437 388 407 352 436 389 0 172 415 322 424 342 427 335 420 338 426 330 429 352 439 325 428 334 423 315 419 337 427 334 436 345 0 401 236 422 330 420 236 405 241 421 247 416 327 410 247 406 349 425 365 417 338 401 277 415 320 30 36 415 295 427 340 431 300 437 319 437 302 446 339 455 318 434 331 429 311 441 336 410 315 429 354 30 48 426 313 427 337 453 331 426 337 440 325 431 355 452 330 411 326 421 310 416 330 408 299 417 313 30 33 415 317 424 347 446 322 420 311 431 316 419 337 450 323 427 334 422 304 425 340 411 299 430 315 30 52 435 333 434 361 448 327 454 365 434 321 432 345 449 326 422 337 408 316 444 364 425 326 436 347 30 42 418 309 428 357 444 331 436 268 422 342 456 334 436 306 428 312 432 327 403 290 402 323 30 68 423 356 407 346 445 361 446 337 433 360 437 379 460 388 432 375 434 368 449 398 398 341 423 373 30 67 44 425 277 434 316 441 274 418 279 432 249 424 318 454 247 441 312 427 241 431 267 380 243 423 270 30 66 46 431 285 426 286 436 252 420 276 437 256 421 292 446 246 439 273 426 252 436 270 411 252 412 243 30 65 45 434 298 422 295 440 276 417 299 430 250 441 301 444 266 436 317 433 268 436 285 417 259 435 298 30 64 47 429 280 428 300 436 273 417 285 436 260 423 293 447 260 417 292 414 243 419 279 424 261 417 269 30 63 43 435 257 429 274 440 239 412 261 441 263 434 305 440 258 416 301 428 254 434 288 397 239 435 303 30 53 417 333 430 359 441 348 428 362 437 362 447 386 433 353 434 368 420 341 439 371 389 333 413 358 30 54 35 432 304 422 327 438 271 423 309 438 294 442 339 424 251 450 310 415 247 439 321 405 236 402 269 30 55 34 433 279 420 308 431 245 424 291 430 276 427 313 449 247 415 289 424 253 446 324 431 267 428 292 30 56 427 365 432 379 436 365 430 377 445 375 432 391 457 377 435 380 437 373 448 390 424 365 423 375 30 57 37 432 301 425 314 436 272 435 313 428 251 440 324 451 258 428 307 414 232 437 325 426 261 421 282 30 38 423 316 421 338 435 321 443 357 440 319 432 334 457 302 426 318 424 282 440 331 400 275 425 326 30 59 39 433 234 430 245 456 241 404 228 433 231 452 291 448 231 419 231 427 261 426 362 423 259 425 346 30 60 40 431 270 433 293 431 262 425 285 429 258 428 299 439 237 413 273 420 265 436 292 397 260 415 272 30 61 41 428 231 436 247 457 265 416 231 440 250 441 299 447 248 412 261 429 229 443 262 420 239 434 260 30 62 431 342 418 355 444 346 445 365 434 342 428 365 454 357 432 367 428 344 449 373 412 353 411 348 30 72 49 429 244 452 252 445 239 420 231 435 243 417 257 436 245 437 254 434 242 403 232 416 237 431 236 30 71 50 428 257 431 285 433 244 412 241 428 243 437 274 436 232 438 230 432 248 419 251 392 244 435 264 30 70 51 439 277 426 264 453 268 416 279 428 243 423 291 426 244 434 283 421 241 409 266 403 241 420 282 30 69 442 378 430 378 452 375 433 378 424 363 442 393 455 377 438 380 429 366 452 389 441 356 430 375 60 34 425 331 436 353 446 237 422 327 431 338 409 351 462 253 378 296 417 227 419 336 432 259 405 347 60 45 422 314 428 340 439 231 417 271 429 261 436 314 459 292 430 328 429 254 417 298 431 236 427 282 
B-1 

60 54 49 440 346 439 313 433 294 435 313 433 298 441 360 462 336 424 334 423 333 453 366 366 311 437 355 60 53 33 434 278 428 295 428 260 412 279 432 268 410 291 441 270 407 282 406 240 428 282 416 255 423 282 60 72 35 414 263 427 327 415 252 415 302 441 267 428 251 418 262 412 284 426 271 433 310 417 260 417 301 60 71 36 425 271 428 320 434 258 404 274 434 247 405 278 440 256 432 283 426 281 431 291 412 259 430 290 60 70 37 419 243 435 318 432 245 429 256 434 266 419 240 442 231 231 235 416 230 427 246 400 237 420 255 60 69 38 433 278 419 328 444 267 428 286 437 248 406 236 461 265 265 278 424 241 409 265 420 249 415 278 60 68 39 431 272 411 296 436 267 416 292 432 261 442 254 462 266 407 267 421 270 414 252 408 240 417 286 60 67 421 352 418 366 442 359 423 365 428 347 418 308 448 360 423 370 424 355 435 382 433 372 456 390 60 55 421 358 416 358 437 346 416 344 431 347 448 390 453 376 433 376 412 327 451 399 399 338 441 389 60 56 40 441 313 441 330 446 427 303 432 277 423 304 447 269 435 304 417 255 418 291 409 254 443 302 60 57 50 432 284 430 300 428 261 409 274 429 272 433 312 452 302 426 299 406 369 453 392 430 364 443 383 60 58 48 433 281 423 293 425 245 408 264 440 235 440 280 437 235 422 267 418 239 407 257 390 250 440 271 60 59 41 431 252 425 284 436 242 428 276 434 244 439 303 446 245 445 294 419 235 420 266 420 255 437 312 60 60 46 437 244 421 250 436 257 423 248 425 230 444 270 437 233 419 237 426 229 420 262 428 261 414 261 60 61 45 438 257 428 313 446 245 415 262 432 391 429 291 444 253 413 287 422 260 422 258 405 233 416 294 60 62 44 428 256 430 309 440 260 412 286 426 372 424 279 440 234 400 250 419 265 416 254 412 241 423 263 60 63 43 426 267 425 314 439 274 418 269 435 372 414 271 446 245 413 250 425 239 403 253 426 251 417 277 60 64 42 430 260 445 327 440 248 414 259 442 385 433 271 455 260 409 275 418 242 429 293 416 258 435 269 60 65 47 423 247 430 284 445 240 411 259 432 393 442 310 450 270 431 311 419 252 413 275 408 259 421 251 60 66 421 341 443 363 452 353 441 373 443 386 423 356 455 303 422 349 425 281 423 361 409 276 430 271 60 58 430 347 447 383 437 361 425 369 441 371 418 366 459 381 439 385 434 375 451 397 429 376 423 374 125 57 34 424 243 435 287 441 241 417 251 430 236 422 268 443 245 432 291 428 245 421 265 417 256 418 252 125 56 36 428 265 430 299 432 261 408 282 423 261 421 298 443 243 438 287 414 243 431 281 408 246 438 295 125 55 35 431 243 414 264 445 232 417 249 438 250 412 287 452 262 410 291 412 244 426 273 422 236 412 253 125 54 37 429 290 417 299 435 267 415 286 435 289 432 305 444 246 415 285 422 261 427 283 426 252 416 276 125 53 423 372 445 400 453 347 432 382 434 374 444 392 460 388 430 386 428 373 438 384 416 366 422 377 125 71 43 432 254 403 275 438 251 420 275 437 247 424 268 443 236 440 267 422 240 423 277 447 255 428 292 125 72 48 418 260 430 282 428 249 406 262 431 250 423 281 461 249 439 289 420 242 423 258 404 237 437 291 125 59 39 434 278 443 294 427 269 422 297 434 311 442 321 441 288 410 309 415 289 442 331 417 285 428 323 125 60 40 417 254 432 294 429 245 414 259 435 265 427 320 451 271 439 285 415 266 428 259 414 247 445 272 125 61 41 435 251 404 262 414 252 397 254 426 236 419 266 440 248 428 285 421 243 425 280 418 261 432 260 125 62 42 415 262 402 288 426 267 407 279 444 283 420 300 448 271 418 288 429 266 423 289 427 275 426 310 125 63 47 421 289 415 300 435 263 426 285 434 294 426 314 453 248 438 302 423 245 429 291 409 266 440 303 125 64 44 414 242 414 252 430 241 419 250 423 241 424 256 439 241 434 241 426 243 420 280 409 244 444 296 125 65 429 365 433 368 449 257 437 364 435 359 449 367 461 356 440 365 433 343 434 352 419 340 433 360 125 66 50 426 263 432 279 438 237 411 254 435 246 422 368 429 243 394 239 432 233 427 239 412 237 433 247 125 67 51 422 252 429 302 439 248 414 262 423 284 428 311 442 243 405 263 417 258 433 308 429 261 430 274 125 68 49 420 250 412 258 427 233 407 232 436 242 415 279 442 247 431 273 425 238 415 248 416 261 425 253 125 69 45 426 267 428 286 432 237 399 232 433 230 437 260 445 251 428 287 430 255 430 280 425 244 402 263 125 70 46 429 280 432 294 433 269 420 289 437 269 417 289 444 247 444 275 422 243 449 298 395 233 416 265 125 38 422 284 413 290 409 251 399 293 436 311 445 350 461 307 420 306 428 273 426 316 436 266 418 274 125 52 414 273 425 342 414 259 402 304 430 308 417 284 450 309 424 305 420 266 432 310 437 276 430 297 125 33 420 314 421 336 442 337 441 360 438 335 428 345 453 331 425 336 432 321 433 317 442 319 448 351 250 53 50 424 291 428 314 428 278 418 303 432 319 444 323 444 272 432 296 428 288 434 305 412 254 429 303 250 54 49 419 247 413 284 438 282 403 288 435 275 410 284 430 254 431 292 415 262 424 275 411 234 413 257 250 55 35 419 261 415 269 404 243 421 266 428 247 414 255 427 247 430 277 420 236 403 248 402 256 425 265 250 56 414 309 433 359 446 335 431 346 436 358 444 368 455 323 426 364 430 360 444 364 419 353 424 335 250 57 33 417 259 419 285 419 250 418 280 427 265 427 295 441 256 437 292 412 250 437 290 414 258 416 275 250 58 37 423 285 420 279 421 245 391 262 427 244 417 276 424 237 437 285 413 238 436 279 401 234 421 255 250 59 38 418 260 408 295 423 255 408 278 433 285 416 321 437 273 426 299 411 272 421 265 408 240 413 267 250 60 39 426 259 418 278 429 244 417 273 429 249 416 279 436 239 439 280 417 243 420 293 413 247 417 277 250 61 40 419 430 315 433 274 410 299 427 289 435 328 453 271 430 297 423 278 413 310 401 270 421 315 
B-1 

250 62 36 417 251 413 270 435 249 406 280 429 242 413 287 436 248 434 320 422 275 415 313 412 242 405 261 250 69 46 417 231 402 250 427 239 405 243 432 244 417 241 425 230 411 240 407 231 410 231 399 235 417 231 250 70 45 436 286 421 316 442 280 424 289 429 270 416 318 443 250 414 280 408 248 440 297 425 354 414 272 250 71 43 424 249 431 300 444 272 411 290 430 259 415 312 407 293 414 295 415 271 426 311 425 289 441 313 250 72 422 358 432 371 447 365 430 368 433 365 440 379 452 362 434 367 407 345 433 356 428 358 421 352 250 63 429 365 440 389 441 249 402 268 424 271 411 299 396 244 434 297 407 243 435 387 439 247 428 391 250 64 52 427 232 414 266 435 247 424 280 430 248 415 273 410 243 439 290 428 229 415 258 428 236 419 242 250 65 51 432 279 426 288 419 376 436 368 420 385 431 432 364 431 387 403 355 433 287 432 385 436 275 250 66 41 425 290 442 302 443 243 411 260 434 263 408 279 423 250 416 276 419 246 432 284 430 254 409 282 250 67 48 414 231 418 265 440 253 436 268 434 248 413 274 409 231 431 268 411 236 434 263 410 241 406 241 250 68 425 334 431 367 443 348 433 361 428 343 423 347 434 350 427 352 418 346 437 368 439 345 435 362 250 34 420 291 405 318 432 295 402 302 427 312 423 445 315 415 314 417 310 432 299 397 299 414 316 250 44 420 260 417 346 443 310 442 344 430 261 419 452 293 409 304 412 301 425 250 421 337 442 301 250 42 423 315 430 343 447 345 439 353 435 327 415 321 443 331 420 321 423 322 419 314 409 328 424 339 250 51 424 289 429 314 442 289 420 300 434 286 429 330 452 252 391 302 425 253 411 265 415 321 451 338 

P generation egg production

DOSE ID 4/20 4/21 4/22 4/23 4/24 4/25 4/26 

week 6 4/27 4/28 4/29 4/30 5/1 5/2 5/3 

week 7 5/4 5/5 5/6

5/

7

5/

8 5/9 5/10 5/11 5/12 5/

0 144 1 0 142 1 1 1 1 0 140 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 0 139 0 141 1 1 1 1 1 0 143 1 1 2 1 1 1 1 1 1 1 1 0 150 1 1 1 1 1 1 1 0 149 1 1 1 1 1 1 1 1 0 148 1 1 1 1 0 147 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 146 
B-1 

0 145 1 1 1 1 1 1 1 1 1 1 1 0 137 2 1 1 1 1 1 1 1 1 0 135 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 133 1 1 1 1 1 1 1 0 134 0 136 1 1 1 1 1 1 1 1 1 0 138 1 2 2 1 1 1 1 1 1 1 1 0 151 0 152 

30 49 1 1 1 1 1 1 1 1 1 1 1 1 1 30 50 1 30 51 1 1 1 30 52 1 1 1 1 1 1 1 1 1 1 1 1 1 30 33 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 30 34 1 1 2 1 2 1 1 1 1 1 1 1 1 30 35 30 36 1 2 1 1 1 1 1 1 30 37 30 38 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 39 2 1 1 1 1 1 1 1 1 30 40 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 41 1 1 1 1 1 1 1 1 1 1 1 1 1 30 42 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 30 43 2 1 1 1 1 1 1 1 1 30 44 1 1 1 1 1 1 1 1 30 45 30 46 30 47 30 48 1 1 1 1 1 1 60 33 1 1 1 1 1 1 1 1 1 1 60 34 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 35 60 36 60 37 1 1 2 
B-1 

60 38 1 1 1 1 1 60 39 1 1 2 1 1 1 1 1 1 1 1 1 60 40 60 47 1 1 1 1 1 1 1 1 1 1 2 1 60 48 1 1 1 1 1 1 1 1 60 49 60 50 2 1 1 2 1 1 1 1 1 1 1 1 1 60 51 60 52 1 1 1 1 1 1 1 1 1 1 1 60 41 1 1 1 1 1 1 60 42 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 60 43 1 1 1 1 1 1 1 1 1 60 44 1 2 1 1 1 1 1 1 1 1 1 60 45 1 1 60 46 1 1 1 1 2 1 1 1 1 1 1 1 1 

125 38 1 1 1 1 1 1 125 39 1 1 1 1 1 1 1 125 37 1 1 125 36 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 35 1 1 1 1 1 1 1 1 1 125 34 1 1 1 1 1 2 1 1 1 1 1 1 1 125 33 1 1 1 1 1 125 44 1 1 2 1 1 1 1 1 1 1 1 125 43 1 1 1 1 1 1 1 125 42 125 41 1 125 40 1 1 1 1 1 1 1 1 1 1 1 1 125 45 1 1 1 1 1 125 46 1 1 1 1 1 1 1 1 1 125 47 1 1 1 1 1 1 1 1 125 48 1 1 1 2 1 1 1 1 1 1 1 1 1 125 49 1 1 1 1 1 1 1 1 1 1 1 1 125 50 1 1 1 1 1 1 1 1 1 125 52 1 1 1 1 1 1 1 1 
B-1 

125 51 1 1 1 1 250 43 1 1 1 1 1 1 1 1 1 1 1 1 1 250 44 1 1 1 1 1 1 1 1 250 45 1 1 250 46 1 1 1 1 1 1 1 1 1 1 1 1 1 250 47 2 1 1 1 1 1 1 1 1 250 48 1 1 1 1 1 1 1 1 1 250 49 1 1 1 1 1 1 1 250 50 250 51 1 1 1 1 1 1 1 1 250 52 1 1 1 1 1 1 1 1 2 1 250 36 1 1 1 1 1 1 1 1 3 1 1 1 1 1 1 1 1 1 1 250 35 1 1 2 1 1 1s 250 34 1 1 2 1 1 1 1 1 1 1 1 250 33 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 250 37 1 1 1 1 250 38 1 1 1 250 39 1 1 1 250 40 250 41 1 1 1 1 1 250 42 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

DOSE ID 5/18 5/19 5/20 5/21 5/22 5/23 5/24 

week 10 5/25 5/26 5/27 5/28 5/29 5/30 5/31 

week 11 6/1 6/2 6/3 6/4 6/5 6/6 6/7 

week 12 6/8 6/9 6/10 

0 144 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 142 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 140 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 0 139 0 141 1 1 1 1 1 1 1 1 1 2 1 1 1 1 0 143 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 150 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 149 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 148 2 1 1 1 1 1 1 1 1 1 1 0 147 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 0 146 1 
B-1 

0 145 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 137 1 1 1 1 1 1 1 1 1 1 0 135 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 133 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 0 134 0 136 1 1 1 1 1 1 1 1 1 1 1 0 138 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 0 151 0 152 

30 49 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 50 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 30 51 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 52 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 33 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 34 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 30 35 1 1 1 1 1 1 1 1 1 1 1 1 30 36 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 37 1 1 1 1 1 1 1 1 30 38 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 39 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 30 40 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 30 41 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 42 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 30 43 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 44 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 30 45 1 1 1 1 1 1 30 46 1 1 1 1 2 1 1 1 1 1 30 47 1 1 1 1 1 1 2 1 1 1 1 30 48 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 33 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 60 34 1 1 1 1 1 1 1 1 2 1 1 1 1 60 35 1 1 1 1 1 1 1 1 1 2 1 1 1 60 36 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 37 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 
B-1 

60 38 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 60 39 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 60 40 1 1 1 1 1 1 1 1 60 47 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 48 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 49 60 50 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 51 60 52 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 60 41 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 42 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 43 2 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 44 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 45 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 46 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

125 38 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 39 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 37 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 36 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 35 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 34 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 125 33 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 44 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 43 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 42 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 41 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 40 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 45 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 46 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 47 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 48 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 49 1 2 1 1 1 1 1 1 1 1 2 1 1 1 1 1 125 50 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 125 52 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 
B-1 

125 51 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 2 1 1 1 1 1 250 43 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 44 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 250 45 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 46 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 47 1 1 1 1 1 1 1 1 250 48 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 49 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 50 1 1 1 1 1 1 1 1 1 250 51 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 2 1 1 2 250 52 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 250 36 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 35 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 34 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 33 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 250 37 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 38 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 39 1 1 1 1 1 1 1 1 2 1 1 250 40 250 41 1 1 1 1 1 1 1 1 2 1 1 1 1 1 250 42 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

DOSE ID 6/15 6/16 6/17 6/18 6/19 6/20 6/21 

week 14 6/22 6/23 6/24 6/25 6/26 6/27 6/28 

week 15 ### ### 7/1 7/2 7/3 7/4 7/5 

week 16 

0 144 1 1 1 1 1 1 1 1 1 0 142 1 1 1 1 1 1 1 1 1 1 0 140 1 1 1 1 1 1 1 1 1 1 1 1 0 139 0 141 1 1 1 1 1 1 1 2 1 1 1 2 0 143 1 1 1 1 1 1 1 1 1 1 1 1 0 150 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 149 1 1 1 1 1 1 1 1 1 1 1 1 0 148 1 1 1 1 1 1 1 1 1 0 147 1 1 1 1 1 1 1 1 2 1 1 0 146 1 1 1 1 1 1 1 
B-1 

0 145 1 1 1 1 1 1 1 1 1 0 137 1 1 1 1 1 1 1 1 2 2 1 1 1 1 1 1 1 0 135 1 1 1 1 1 1 1 1 1 1 1 1 0 133 1 1 1 1 1 1 1 1 1 1 2 1 1 0 134 0 136 1 1 1 0 138 1 1 1 1 1 1 1 1 1 1 2 1 0 151 0 152 

30 49 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 50 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 30 51 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 52 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 33 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 34 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 35 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 30 36 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 37 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 38 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 39 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 40 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 41 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 42 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 43 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 44 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 30 45 1 1 1 1 1 1 1 2 1 1 1 1 1 30 46 1 1 1 1 2 1 1 1 1 1 2 1 1 1 1 1 1 30 47 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 30 48 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 33 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 34 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 60 35 1 1 1 1 1 1 1 1 1 1 2 2 1 1 1 2 1 1 1 1 60 36 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 60 37 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 
B-1 

60 38 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 39 1 1 1 1 1 1 1 1 1 2 1 1 1 60 40 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 60 47 1 1 1 1 1 1 1 1 1 1 60 48 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 49 60 50 2 1 1 1 60 51 60 52 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 41 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 60 42 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 60 43 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 44 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 60 45 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 46 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 

125 38 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 125 39 1 1 2 1 1 125 37 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 36 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 35 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 34 1 1 1 1 1 1 1 1 1 2 1 1 1 2 1 125 33 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 125 44 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 43 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 125 42 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 125 41 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 125 40 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 45 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 46 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 125 47 1 1 1 1 2 1 1 1 1 1 1 125 48 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 125 49 1 1 1 1 1 1 1 1 1 2 1 1 1 2 1 1 1 1 125 50 2 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 125 52 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 
B-1 

125 51 1 1 1 1 1 1 2 1 1 1 2 1 1 1 250 43 1 1 1 1 1 1 1 250 44 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 45 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 46 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 250 47 250 48 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 49 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 50 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 250 51 2 1 1 1 1 1 1 2 2 2 2 1 1 1 1 1 1 2 1 2 1 250 52 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 250 36 1 1 1 1 1 1 1 1 1 1 1 2 1 250 35 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 250 34 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 33 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 250 37 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 38 1 1 1 1 1 1 1 1 1 1 1 250 39 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 40 250 41 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 42 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 
DOSE ID 7/6 7/7 7/8 7/9 7/10 7/11 7/12 

week 

17 7/13 7/14 7/15 7/16 7/17 7/18 7/19 7/20 ### ### ### 

week 

18 

0 144 1 1 1 1 1 1 1 1 

0 142 1 1 1 1 1 1 1 1 1 1 

0 140 1 1 1 1 1 1 2 1 1 1 1 

0 139 

0 141 1 1 2 1 1 1 1 

0 143 1 1 1 1 1 1 1 1 1 1 

0 150 1 1 1 2 

0 149 1 1 1 1 1 1 

0 148 1 2 1 

0 147 1 1 1 1 

0 146 1 1 1 1 1 2 1 1 1 

0 145 1 1 1 1 1 1 1 1 

0 137 1 2 1 2 1 1 1 1 1 

0 135 1 1 1 1 1 

0 133 1 1 1 1 1 1 1 2 

0 134 

0 136 1 1 

0 138 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0 151 

0 152 

30 49 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30 50 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 

30 51 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30 52 1 1 1 1 1 1 1 1 1 1 1 1 1 

30 33 1 1 1 1 1 1 1 2 1 1 1 1 2 1 1 1 1 

30 34 1 1 1 1 1 1 1 2 1 1 1 1 1 1 

30 35 1 1 1 1 1 2 1 1 1 1 1 1 

30 36 1 1 1 2 1 1 1 1 2 1 

30 37 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30 38 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30 39 1 1 1 1 2 1 2 1 1 1 1 1 1 1 1 1 

30 40 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30 41 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 

30 42 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30 43 1 1 1 1 1 1 1 1 1 1 1 1 

30 44 1 1 1 1 1 1 2 1 1 1 1 1 1 

30 45 1 1 1 1 2 1 1 1 1 1 1 1 

30 46 1 1 1 1 1 1 1 2 1 1 1 1 1 1 

30 47 1 1 1 1 1 1 1 2 1 1 1 1 1 1 

30 48 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 

60 33 1 1 1 1 1 1 2 1 1 1 1 1 1 

60 34 1 1 1 1 1 1 1 1 1 1 2 1 1 1 

60 35 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60 36 1 1 1 1 1 1 1 1 1 1 1 1 1 

60 37 1 1 1 1 1 1 2 1 1 1 1 1 1 
B-1 

60 38 1 1 1 1 1 

60 39 1 1 1 1 1 2 1 1 1 1 1 1 1 

60 40 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60 47 1 1 1 1 1 1 1 2 1 1 1 1 1 

60 48 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 

60 49 

60 50 1 1 1 1 2 1 2 1 1 1 1 1 1 1 1 1 

60 51 

60 52 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 

60 41 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60 42 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 

60 43 1 1 1 1 1 1 1 1 1 1 1 1 

60 44 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 

60 45 1 1 1 1 1 1 1 2 1 1 1 1 1 2 1 1 1 

60 46 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 

125 38 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 

125 39 1 

125 37 1 1 1 1 1 1 1 1 1 1 1 1 

125 36 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

125 35 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 

125 34 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 

125 33 1 1 1 1 1 1 1 2 1 1 1 1 2 1 

125 44 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 

125 43 1 1 1 1 1 1 1 1 1 1 2 1 1 2 1 1 1 

125 42 1 1 1 1 1 1 2 1 1 1 1 

125 41 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

125 40 1 1 1 1 1 1 1 1 1 1 1 1 2 1 

125 45 1 1 1 1 1 1 1 1 1 1 1 

125 46 1 1 1 1 1 1 1 1 1 1 2 2 1 1 1 1 

125 47 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 

125 48 1 1 1 1 1 1 2 1 1 1 1 1 1 1 

125 49 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 

125 50 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

125 52 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 

125 51 1 1 1 1 1 1 1 2 1 1 1 1 1 1 

250 43 1 1 1 1 1 1 1 

250 44 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 

250 45 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

250 46 1 1 1 1 1 2 1 1 1 1 1 1 1 

250 47 1 1 1 1 1 1 

250 48 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

250 49 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 

250 50 1 1 1 1 1 1 1 1 1 2 1 1 1 1 

250 51 2 2 1 1 1 1 2 1 1 1 1 1 1 2 1 

250 52 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 

250 36 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 

250 35 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 

250 34 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 
B-1 

250 33 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

250 37 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

250 38 1 1 2 1 1 1 1 

250 39 1 1 1 1 2 1 1 1 1 1 

250 40 

250 41 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

250 42 1 1 2 1 1 1 1 1 1 1 2 1 1 

F1 clocaa measurements and foam 

8/3/2009 8/11/2009 

DOSE ID 

cg 

area foam DOSE ID 

cg 

area foam DOSE ID first foam hatch date day of onset 

0 218 168 Y 0 221 195 Y 0 205 8/3 6/26 38 

0 221 132 Y 0 218 182 Y 0 206 8/3 6/26 38 

0 279 144 Y 0 236 72 N 0 213 8/3 6/26 38 

0 213 210 Y 0 224 72 N 0 214 8/3 6/26 38 

0 214 210 Y 0 216 143 Y 0 216 8/10 6/28 43 

0 232 110 Y 0 227 289 Y 0 218 8/3 6/26 38 

0 206 143 Y 0 280 132 N 0 220 8/3 6/26 38 

0 244 156 Y 0 240 255 Y 0 221 8/3 6/27 37 

0 205 156 Y 0 222 72 N 0 222 8/19 6/26 54 

0 246 130 Y 0 229 81 N 0 224 9/16 6/26 82 

0 224 110 N 0 243 256 Y 0 227 8/3 6/26 38 

0 236 72 N 0 220 255 Y 0 229 8/17 6/26 52 

0 227 221 Y 0 213 342 Y 0 232 8/3 6/28 36 

0 216 90 N 0 278 210 Y 0 236 8/19 6/26 54 

0 240 168 Y 0 214 210 Y 0 240 8/3 6/26 38 

0 280 90 N 0 232 195 Y 0 243 8/3 6/26 38 

0 229 35 N 0 244 240 Y 0 244 8/3 6/25 39 

0 222 100 N 0 206 180 Y 0 246 8/3 6/26 38 

0 243 154 Y 0 246 132 Y 0 279 8/3 6/29 35 

0 220 195 Y 0 205 272 Y 0 280 8/16 6/29 48 

30 255 108 Y 30 257 16814 Y 30 204 9/16 6/26 82 

30 257 154 Y 30 255 208 Y 30 205 9/16 6/26 82 

30 246 144 Y 30 246 323 Y 30 209 9/16 6/26 82 

30 237 54 N 30 237 154 Y 30 213 9/16 6/27 81 

30 238 100 Y 30 325 100 N 30 215 9/16 6/26 82 

30 236 25 N 30 282 196 Y 30 216 8/3 6/26 38 

30 223 196 Y 30 223 210 Y 30 219 8/3 6/26 38 

30 244 25 N 30 244 121 Y 30 223 8/3 6/26 38 

30 215 100 N 30 215 196 Y 30 224 8/3 6/25 39 

30 205 25 N 30 205 80 N 30 233 8/3 6/26 38 

30 325 25 N 30 224 195 Y 30 234 8/3 6/25 39 

30 282 90 Y 30 233 169 Y 30 237 8/10 6/26 45 

30 233 108 Y 30 219 240 Y 30 240 9/16 6/27 81 

30 224 120 Y 30 248 132 Y 30 242 9/16 6/26 82 

30 213 81 N 30 216 240 Y 30 244 8/10 6/27 44 

30 219 121 Y 30 217 25 N 30 246 8/3 6/25 39 

30 216 143 Y 30 240 132 N 30 255 8/3 6/27 37 
B-1 

30 234 132 Y 30 272 54 N 30 257 8/3 6/26 38 

30 240 72 N 30 204 100 N 30 282 8/3 6/29 35 

30 272 25 N 30 209 40 Y 30 325 9/16 6/26 82 

30 204 90 N 60 176 168 Y 60 177 8/7 6/26 42 

30 209 25 N 60 167 132 Y 60 109 8/3 6/26 38 

60 176 169 Y 60 116 210 Y 60 111 8/3 6/28 36 

60 167 130 Y 60 111 234 Y 60 114 8/3 6/29 35 

60 111 210 Y 60 164 144 Y 60 116 8/3 6/27 37 

60 116 143 Y 60 151 182 Y 60 117 8/3 6/27 37 

60 151 144 Y 60 114 156 Y 60 120 8/7 6/26 42 

60 164 100 N 60 187 289 Y 60 136 9/1 6/29 64 

60 114 100 Y 60 190 64 N 60 151 8/3 6/28 36 

60 187 192 Y 60 178 169 Y 60 161 9/16 6/26 82 

60 190 120 N 60 177 143 Y 60 164 9/16 6/28 80 

60 178 104 Y 60 180 240 Y 60 167 8/3 6/28 36 

60 177 100 N 60 161 80 N 60 169 8/3 6/29 35 

60 180 168 Y 60 109 225 Y 60 171 8/3 6/25 39 

60 109 195 Y 60 173 238 Y 60 173 8/3 6/25 39 

60 161 143 N 60 120 256 Y 60 176 8/3 7/4 30 

60 120 156 N 60 169 256 Y 60 178 8/3 6/28 36 

60 173 240 Y 60 171 168 Y 60 180 8/3 6/26 38 

60 171 210 Y 60 117 154 Y 60 187 8/3 6/29 35 

60 169 195 Y 60 136 36 N 60 190 8/26 6/26 61 

60 117 169 Y 125 110 224 Y 125 109 8/11 6/29 43 

60 136 25 N 125 109 81 Y 125 114 8/3 6/28 36 

125 109 25 N 125 184 210 Y 125 118 8/3 6/27 37 

125 114 168 Y 125 193 208 Y 125 119 8/11 6/25 47 

125 193 25 N 125 182 240 Y 125 131 8/3 6/25 39 

125 184 144 N 125 155 169 Y 125 134 8/3 6/25 39 

125 155 25 N 125 170 165 Y 125 137 8/3 6/26 38 

125 182 110 Y 125 106 210 Y 125 151 8/3 6/26 38 

125 185 168 Y 125 97 224 Y 125 152 9/16 6/29 79 

125 170 108 Y 125 188 48 N 125 155 8/10 6/26 45 

125 188 25 N 125 133 169 Y 125 157 9/16 6/26 82 

125 189 120 Y 125 151 180 Y 125 161 8/3 6/25 39 

125 151 90 Y 125 183 225 Y 125 170 8/3 6/26 38 

125 134 165 Y 125 161 169 Y 125 182 8/3 6/29 35 

125 161 90 Y 125 131 196 Y 125 183 8/3 6/25 39 

125 183 182 Y 125 118 270 Y 125 184 8/9 6/26 44 

125 118 195 Y 125 157 56 N 125 185 8/3 6/28 36 

125 131 156 Y 125 152 70 N 125 188 9/16 6/28 80 

125 152 70 N 125 134 224 Y 125 189 8/3 6/25 39 

125 157 63 N 125 119 132 N 125 193 8/10 6/28 43 

125 119 80 N 250 158 132 Y 250 103 8/3 6/25 39 

125 137 180 Y 250 141 144 N 250 105 8/7 6/27 41 

250 135 196 Y 250 138 120 Y 250 106 8/26 6/25 62 

250 118 70 N 250 111 210 Y 250 108 8/3 6/26 38 

250 120 70 N 250 105 169 Y 250 109 8/3 6/28 36 
B-1 

250 140 195 Y 250 112 306 Y 250 111 8/9 6/25 45 

250 255 56 N 250 135 156 Y 250 112 8/3 6/26 38 

250 109 154 Y 250 118 110 N 250 116 8/3 6/26 38 

250 116 247 Y 250 140 210 Y 250 118 9/16 6/26 82 

250 131 25 N 250 120 121 N 250 120 9/16 6/26 82 

250 106 132 Y 250 155 56 N 250 131 8/19 6/26 54 

250 132 143 Y 250 109 210 Y 250 132 8/3 6/27 37 

250 158 90 N 250 116 208 Y 250 135 8/3 6/27 37 

250 141 54 N 250 131 42 N 250 138 8/7 6/26 42 

250 111 80 N 250 132 132 Y 250 140 8/3 6/25 39 

250 138 100 N 250 106 100 N 250 141 9/16 6/26 82 

250 105 90 N 250 108 256 Y 250 155 9/16 6/26 82 

250 112 169 Y 250 174 169 Y 250 158 8/11 6/26 46 

250 103 195 Y 250 177 110 N 250 174 8/3 6/30 34 

250 177 35 N 250 103 255 Y 250 177 9/16 6/28 80 

250 108 210 Y 

250 174 80 Y 

F1 egg strength 

Dose ID Thickness 1 Thickness 2 Thickness avg Strength Yolk 

 

0 202 1.03 0.99 1.01 0.395 Y 

0 203 1.28 0.87 1.075 1.223 Y 

0 203 1.23 1.27 1.25 1.509 Y 

0 207 1.06 0.92 0.99 1.004 Y 

0 209 1.1 0.7 0.9 0.543 Y 

0 209 0.77 0.8 0.785 0.999 Y 

0 211 0.75 0.96 0.855 0.882 Y 

0 212 1.06 0.78 0.92 1.145 Y 

0 212 0.79 0.67 0.73 0.739 Y 

0 215 1.15 1.35 1.25 0.547 Y 

0 215 0.92 0.78 0.85 0.87 Y 

0 217 0.85 0.92 0.885 0.946 Y 

0 219 0.56 0.8 0.68 1.038 Y 

0 221 0.72 0.88 0.8 1.283 Y 

0 225 0.6 0.81 0.705 1.196 Y 

0 225 1.35 1.3 1.325 1.28 Y 

0 226 0.72 1.24 0.98 1.09 N 

0 226 1.18 0.98 1.08 1.052 Y 
B-1 

0 226 0.76 0.84 0.8 0.503 Y 

0 228 0.77 0.72 0.745 0.699 Y 

0 228 1.02 0.93 0.975 0.988 Y 

0 234 1.02 0.81 0.915 1.105 Y 

0 234 1.55 1.2 1.375 0.976 Y 

0 235 1.12 1.45 1.285 0.893 Y 

0 235 1.34 1.6 1.47 1.014 Y 

0 237 1 0.87 0.935 0.908 Y 

0 237 1.15 1.35 1.25 0.974 Y 

0 242 0.34 0.22 0.28 1.027 Y 

0 242 0.71 0.82 0.765 1.206 Y 

0 245 0.59 0.77 0.68 1.163 Y 

0 299 0.41 0.33 0.37 1.063 N 

30 202 0.94 0.99 0.965 1.455 Y 

30 206 0.85 0.8 0.825 0.472 Y 

30 211 0.68 0.55 0.615 0.426 Y 

30 211 0.39 0.45 0.42 0.811 Y 

30 212 1 0.82 0.91 1.023 Y 

30 220 0.73 0.67 0.7 1.117 Y 

30 220 1.16 1.12 1.14 0.944 Y 

30 221 0.31 0.4 0.355 1.471 Y 

30 225 0.85 0.93 0.89 0.438 Y 

30 226 1.21 0.78 0.995 0.86 Y 

30 227 1.04 1.43 1.235 1.15 Y 

30 230 1.16 0.84 1 1.097 Y 

30 230 0.59 0.75 0.67 1.122 Y 

30 231 0.95 0.9 0.925 1.181 Y 

30 232 0.61 0.49 0.55 1.014 Y 

30 232 1.07 1.03 1.05 0.808 N 

30 232 0.73 0.77 0.75 0.811 Y 

30 235 0.91 0.83 0.87 1.192 Y 

30 235 1.15 1.03 1.09 1.449 Y 

30 242 1.21 0.85 1.03 0.97 Y 

30 242 1.23 1.3 1.265 1.239 Y 

30 247 1.13 1.03 1.08 1.093 Y 

30 247 0.43 0.34 0.385 1.018 Y 

30 249 0.41 0.42 0.415 1.301 Y 

30 250 1.05 0.84 0.945 0.359 Y 

30 250 0.64 0.58 0.61 1.068 Y 

30 251 1.27 0.75 1.01 1.431 Y 

30 253 0.41 0.55 0.48 1.021 N 

30 253 1.22 1.14 1.18 0.98 Y 

30 253 1.04 0.92 0.98 1.002 Y 

60 102 0.58 0.4 0.49 1.265 Y 

60 102 1.34 1.58 1.46 0.918 Y 

60 110 0.4 0.32 0.36 1.54 Y 

60 115 0.84 0.82 0.83 1.027 Y 

60 115 0.59 0.48 0.535 0.813 Y 
B-1 

60 119 0.88 0.63 0.755 1.074 Y 

60 165 0.79 0.92 0.855 1.002 Y 

60 165 0.89 1.2 1.045 1.173 Y 

60 166 0.96 1.03 0.995 0.998 Y 

60 170 0.93 0.99 0.96 1.239 Y 

60 170 0.73 0.63 0.68 1.161 Y 

60 172 0.68 0.87 0.775 0.975 Y 

60 172 0.92 0.96 0.94 1.21 Y 

60 174 1.03 0.94 0.985 0.952 N 

60 174 0.5 0.75 0.625 0.477 Y 

60 179 0.7 0.62 0.66 0.812 Y 

60 179 0.61 0.53 0.57 0.815 Y 

60 179 1.38 1.25 1.315 0.996 Y 

60 181 0.75 0.64 0.695 0.882 Y 

60 181 0.72 0.89 0.805 0.96 Y 

60 183 1.2 1.44 1.32 0.945 Y 

60 184 0.5 0.44 0.47 1.269 Y 

60 184 0.95 0.81 0.88 1.408 Y 

60 185 1.07 0.98 1.025 1.111 Y 

60 185 1.18 0.69 0.935 1.005 Y 

60 189 0.48 0.49 0.485 1.225 Y 

60 189 0.89 0.7 0.795 1.12 Y 

125 101 Y 

125 101 0.38 0.33 0.355 1.383 N 

125 101 0.47 0.34 0.405 1.482 N 

125 101 1 1.21 1.105 1.767 Y 

125 103 0.95 1.02 0.985 1.043 Y 

125 106 1.05 0.8 0.925 0.817 N 

125 106 0.62 0.97 0.795 0.93 Y 

125 107 Y 

125 107 0.35 0.34 0.345 1.185 N 

125 107 0.79 0.72 0.755 1.243 Y 

125 110 0.67 0.51 0.59 0.997 N 

125 111 0.39 0.32 0.355 1.159 N 

125 111 0.72 0.68 0.7 0.973 Y 

125 112 0.87 0.91 0.89 0.591 Y 

125 112 0.61 0.53 0.57 1.277 N 

125 113 Y 

125 113 1.06 0.8 0.93 1.187 N 

125 113 1.1 1.22 1.16 0.838 Y 

125 114 0.97 0.88 0.925 1.121 N 

125 117 0.97 0.99 0.98 1.296 Y 

125 117 0.67 0.81 0.74 1.305 Y 

125 119 1.09 0.76 0.925 1.292 Y 

125 135 Y 

125 135 0.9 0.86 0.88 1.364 N 

125 135 0.65 0.88 0.765 1.126 Y 

125 156 0.72 0.56 0.64 1 N 
B-1 

125 156 0.8 0.89 0.845 1.018 Y 

125 158 0.69 0.64 0.665 0.408 N 

125 158 0.8 0.72 0.76 0.993 Y 

125 159 Y 

125 159 0.55 0.32 0.435 0.668 N 

125 159 0.71 0.96 0.835 0.76 Y 

125 160 0.89 0.91 0.9 0.695 N 

125 160 0.79 0.97 0.88 0.937 Y 

125 163 0.85 0.82 0.835 1.278 N 

125 163 0.76 0.81 0.785 0.999 Y 

125 165 Y 

125 165 0.53 0.5 0.515 1.185 N 

125 165 0.7 0.75 0.725 1.371 Y 

125 166 0.75 0.65 0.7 0.403 N 

125 167 0.6 0.57 0.585 0.814 N 

125 167 1.02 1.21 1.115 1.132 Y 

125 181 Y 

125 181 0.67 0.65 0.66 1.153 N 

125 181 0.77 0.88 0.825 1.283 Y 

125 191 0.77 0.7 0.735 1.114 Y 

125 191 0.78 0.99 0.885 1.414 N 

125 191 0.66 0.79 0.725 1.314 Y 

250 102 0.89 0.85 0.87 1.524 Y 

250 102 0.76 0.68 0.72 1.006 Y 

250 107 0.34 0.32 0.33 0.794 N 

250 109 1.21 0.85 1.03 1.344 N 

250 110 Y 

250 110 0.31 0.35 0.33 1.363 N 

250 110 0.51 0.42 0.465 0.976 Y 

250 114 Y 

250 114 0.63 0.54 0.585 0.918 N 

250 114 0.85 0.89 0.87 1.153 Y 

250 115 0.39 0.46 0.425 1.298 Y 

250 115 0.57 0.53 0.55 1.658 Y 

250 122 0.8 0.68 0.74 1.275 N 

250 131 0.77 0.86 0.815 0.811 N 

250 133 0.61 0.46 0.535 1.164 N 

250 134 0.39 0.36 0.375 0.8 N 

250 136 0.59 0.61 0.6 1.01 Y 

250 136 0.77 0.76 0.765 1.191 N 

250 151 0.66 0.65 0.655 1.217 Y 

250 151 0.41 0.45 0.43 1.396 Y 

250 152 1.05 0.86 0.955 1.204 Y 

250 152 0.35 0.27 0.31 1.254 Y 

250 153 0.74 0.7 0.72 1.257 N 

250 153 0.88 0.75 0.815 0.927 Y 

250 154 0.7 0.69 0.695 0.673 Y 

250 156 Y 
B-1 

250 156 0.49 0.46 0.475 1.394 N 

250 156 0.66 0.59 0.625 1.456 Y 

250 159 Y 

250 159 0.58 0.56 0.57 1.282 N 

250 159 0.61 0.68 0.645 1.382 Y 

250 161 Y 

250 161 0.85 0.94 0.895 1.307 N 

250 161 0.46 0.44 0.45 1.257 Y 

250 171 Y 

250 171 0.39 0.28 0.335 1.389 N 

250 171 0.9 0.92 0.91 1.485 Y 

250 193 1.14 0.86 1 0.943 N 

F1 sperm measurements 

DOSE ID % motile % progressive sperm/gram 

0 214 24 0 690.9 

0 214 19 3 597.5 

0 214 21 0 728.2 

0 218 12 0 295.8 

0 218 22 0 254.5 

0 218 18 0 385.2 

0 220 22 6 427.8 

0 220 44 4 641.6 

0 220 18 0 451.5 

0 221 32 0 228.7 

0 221 25 0 465.6 

0 221 7 0 220.8 

0 222 29 8 606 

0 222 5 0 451.5 

0 222 15 0 796.1 

0 227 77 13 767.9 

0 227 29 8 316.6 

0 227 42 5 310.4 

0 229 11 0 558.5 

0 229 6 0 606 

0 229 5 0 665.4 

0 233 45 7 796.1 

0 233 52 3 392.1 

0 233 44 6 404 

0 240 67 9 401.4 

0 240 63 8 550.8 

0 240 56 11 532.2 

0 279 91 0 143.8 
B-1 

0 279 67 0 196.1 

0 279 82 0 143.8 

0 280 84 22 344 

0 280 63 5 275.2 

0 280 63 11 240.8 

30 205 0 0 13.1 

30 205 50 0 52.3 

30 205 0 0 39.2 

30 215 63 2 453.1 

30 215 68 4 601.2 

30 215 61 3 313.7 

30 217 33 5 274.5 

30 217 58 11 248.3 

30 217 48 14 274.5 

30 219 19 0 104.5 

30 219 10 0 963.9 

30 219 7 7 718.9 

30 223 30 4 252.1 

30 223 25 0 186.7 

30 223 35 0 158.7 

30 237 47 4 508.3 

30 237 36 0 428.4 

30 237 23 2 472 

30 244 3 0 246.9 

30 244 12 0 181.5 

30 244 8 0 188.8 

30 248 38 8 308.9 

30 248 44 8 297.1 

30 248 27 0 261.4 

30 255 19 2 364.6 

30 255 30 4 392.1 

30 255 13 0 371.5 

60 111 49 0 285.9 

60 111 59 5 514.6 

60 111 64 10 702.5 

60 114 50 4 332.7 

60 114 57 2 499.1 

60 114 40 16 297.1 

60 116 73 8 355.8 

60 116 70 8 442.9 

60 116 56 5 428.4 

60 120 71 16 233.4 

60 120 56 6 298.8 

60 120 56 4 233.4 
B-1 

60 136 35 2 562 

60 136 33 5 562 

60 136 37 0 666.6 

60 151 62 15 617.9 

60 151 55 7 499.1 

60 151 53 7 653.5 

60 164 4 0 662.2 

60 164 0 0 810.4 

60 164 4 0 801.7 

60 171 65 16 492.7 

60 171 79 8 241.3 

60 171 67 11 542.9 

60 177 74 7 441.1 

60 177 33 0 392.1 

60 177 34 0 473.8 

60 187 33 2 617.9 

60 187 25 0 606 

60 187 33 0 617.9 

125 109 21 2 575.1 

125 109 15 0 357.3 

125 109 13 0 470.5 

125 119 50 6 348.5 

125 119 65 3 337.7 

125 119 37 3 326.8 

125 131 3 0 413.9 

125 131 30 4 294.1 

125 131 21 5 206.9 

125 134 90 9 632.9 

125 134 79 11 454 

125 134 68 7 385.2 

125 155 69 8 448.1 

125 155 62 0 196.1 

125 155 55 0 186.7 

125 157 67 6 112 

125 157 38 8 80.9 

125 157 14 0 87.1 

125 161 51 1 620.8 

125 161 51 5 792.4 

125 161 56 9 555.5 

125 170 56 4 297.1 

125 170 52 9 522.8 

125 170 48 4 320.8 

125 182 53 2 792.9 

125 182 58 7 766.8 

125 182 51 6 679.7 

125 193 23 5 522.8 

125 193 34 0 558.5 

125 193 41 2 487.2 
B-1 

250 106 55 9 558.5 

250 106 61 14 582.2 

250 106 33 8 570.3 

250 108 61 8 555.5 

250 108 58 5 620.8 

250 108 52 9 708 

250 109 61 6 426.5 

250 109 40 6 240.8 

250 109 39 3 227 

250 116 70 14 597.5 

250 116 64 16 746.9 

250 116 67 13 802.9 

250 131 59 13 246 

250 131 66 19 246 

250 131 44 6 246 

250 132 50 0 294.1 

250 132 69 2 400.3 

250 132 35 4 465.6 

250 135 50 6 426.5 

250 135 49 5 433.4 

250 135 60 10 467.8 

250 158 48 0 297.1 

250 158 30 3 392.1 

250 158 61 6 368.3 

250 174 68 25 748.6 

250 174 68 27 522.8 

250 174 55 13 368.3 

250 177 0 0 0 

250 177 0 0 0 

250 177 0 0 0 
F1 growth measurements 

7/6/2009 7/13/2009 7/20/2009 

DOSE baby id BW tarsus 1 tarsus 2 avg tarsus culmen 1 culmen 2 avg culmen DOSE baby id BW tarsus 1 tarsus 2 avg tarsus culmen 1 culmen 2 avg culmen DOSE baby id BW tarsus 

0 372 8 15.6 15.6 15.6 3.3 3.3 3.3 0 371 33 24.2 23.3 23.75 6 5.8 5.9 0 356 50 190 371 16 18.9 18.5 18.7 3.9 3.7 3.8 0 377 30 21.8 21.9 21.85 5.5 6 5.75 0 365 56 190 387 12 17.7 17.7 17.7 3.5 3.7 3.6 0 382 33 22.3 22.5 22.4 5.4 5.5 5.45 0 385 46 170 362 22 21.3 21.4 21.35 3.7 3.9 3.8 0 387 31 21.9 21.7 21.8 5.4 5.5 5.45 0 232 55 180 377 14 19.1 18.6 18.85 4.1 4.2 4.15 0 312 38 21.6 21.6 21.6 6.5 6.9 6.7 0 204 48 220 384 8 16.4 16.3 16.35 2.9 3.1 3 0 363 38 21.6 22 21.8 6 5.9 5.95 0 231 66 240 356 13 17.8 17.8 17.8 3.8 3.9 3.85 0 362 43 22.4 22.1 22.25 5.8 6 5.9 0 349 69 210 370 15 18.9 19.2 19.05 4.1 4.2 4.15 0 358 27 19.5 19.4 19.45 5.7 6 5.85 0 383 51 210 386 10 19.7 19.7 19.7 4.1 4.1 4.1 0 367 33 25.1 24.8 24.95 6.2 6.3 6.25 0 368 54 200 357 17 16.1 16.2 16.15 3.2 3.2 3.2 0 355 44 25.6 26 25.8 6.5 6.5 6.5 0 358 48 18

30 67 20 21.9 21.9 21.9 4.5 4.4 4.45 30 83 31 21.1 21.3 21.2 7.2 7 7.1 30 96 80 2230 98 19 20.7 20.7 20.7 4.2 4.3 4.25 30 73 40 23.1 22.9 23 6.4 6.3 6.35 30 82 2130 425 7 16.7 16.7 16.7 3.7 3.5 3.6 30 418 31 20.9 20.7 20.8 5.7 5.9 5.8 30 414 2330 442 7 15.4 15.3 15.35 3.1 3.1 3.1 30 417 31 21.1 20.7 20.9 5.9 5.7 5.8 30 432 1630 82 17 20.7 21.2 20.95 4.2 4.2 4.2 30 86 40 19.1 19.7 19.4 6.6 6.3 6.45 30 97 2330 418 13 18 18 18 4.1 4 4.05 30 82 39 23.3 23.6 23.45 6.6 7.3 6.95 30 71 2330 414 16 19.9 20 19.95 3.9 3.8 3.85 30 67 40 22.2 21.6 21.9 6.1 6.8 6.45 30 416 30 416 12 18.1 18 18.05 3.5 3.5 3.5 30 409 40 19.3 19 19.15 6.2 6.5 6.35 30 438 1930 422 13 18.4 18.3 18.35 3.7 3.6 3.65 30 94 41 21.1 21.2 21.15 7 6.9 6.95 30 87 2430 71 18 19.8 19.8 19.8 4.5 4.4 4.45 30 23 36 21.9 22.6 22.25 7.1 7.2 7.15 30 424 2060 283 17 20.6 20.4 20.5 3.6 3.7 3.65 60 510 16 16.6 16.3 16.45 4.8 5.1 4.95 60 262 57 2160 289 16 18.7 19.3 19 4 3.9 3.95 60 296 25 18.4 17.3 17.85 6.7 6.4 6.55 60 268 60 305 7 16.3 16.3 16.3 3.6 3.6 3.6 60 309 17 15.9 15.4 15.65 4.9 5.2 5.05 60 486 2160 249 20 20.3 20.5 20.4 4.5 4.5 4.5 60 282 28 17.8 17.7 17.75 7.4 6.9 7.15 60 266 1960 263 15 18.8 18.9 18.85 3.9 4 3.95 60 223 30 21.3 21.5 21.4 6.2 6.7 6.45 60 298 44 2060 257 22 21.5 21.5 21.5 4 4.1 4.05 60 269 31 21.2 21.5 21.35 7.1 6.9 7 60 300 2160 307 12 18.9 18.5 18.7 2.9 3.3 3.1 60 273 27 20.9 20.6 20.75 6.8 6.3 6.55 60 103 1960 295 16 19.3 19.5 19.4 3.6 3.7 3.65 60 303 27 16.9 17.2 17.05 6.1 5.7 5.9 60 283 2460 300 13 17.9 18 17.95 3.4 3.6 3.5 60 276 37 22.4 22.5 22.45 7.1 7 7.05 60 104 54 19
B-1 

60 253 20 20.3 20.4 20.35 3.8 4 3.9 60 279 29 19.1 20 19.55 6 6.1 6.05 60 250 50 21

125 338 13 17.4 17.3 17.35 3.4 3.5 3.45 125 344 25 18.1 17.1 17.6 5.2 5.6 5.4 125 305 58 21125 341 12 17.3 17.3 17.3 3.5 3.6 3.55 125 329 35 17.8 18.4 18.1 5.2 5.5 5.35 125 104 59 125 343 7 15.3 15.6 15.45 3.6 3.6 3.6 125 252 50 21.2 23.5 22.35 7.2 6.9 7.05 125 269 57 21125 282 23 21.5 21.5 21.5 4.9 4.6 4.75 125 330 29 17.4 17.8 17.6 5.8 6.1 5.95 125 289 60 18125 252 30 23.9 23.8 23.85 4.4 4.5 4.45 125 256 35 22.2 20.9 21.55 6.6 6 6.3 125 288 64 22125 274 23 22.2 22.2 22.2 4.5 4.5 4.5 125 305 32 22.2 22.6 22.4 7.3 7.3 7.3 125 244 66 21125 332 14 19.3 19.3 19.3 3.5 3.4 3.45 125 247 41 21.4 21.5 21.45 6.5 6.5 6.5 125 295 64 21125 313 20 20.8 20.7 20.75 4.1 4.3 4.2 125 314 31 20 20.2 20.1 6.5 6.1 6.3 125 332 51 22125 318 20 20.8 20.7 20.75 4.8 4.7 4.75 125 303 34 20.5 20.6 20.55 6.7 6.2 6.45 125 315 57 20125 289 22 20.9 21 20.95 4.3 4.5 4.4 125 309 36 19.8 19.6 19.7 6.2 6.1 6.15 125 326 53 22250 362 21 21 20.9 20.95 4.7 4.7 4.7 250 367 39 19.2 19.3 19.25 5.9 6.4 6.15 250 336 77 17250 358 23 21.3 21.5 21.4 4.2 4.2 4.2 250 355 41 20.5 20.2 20.35 6.6 6 6.3 250 368 48 21250 384 11 16.4 16.5 16.45 3.9 4.1 4 250 383 35 21.6 21 21.3 6.2 6.5 6.35 250 392 67 18250 406 11 17 17 17 3.4 3.1 3.25 250 360 43 20.8 21 20.9 5.1 5.5 5.3 250 372 53 23250 324 27 21.8 21.8 21.8 4.6 4.7 4.65 250 377 37 21.1 21.5 21.3 5.8 6.1 5.95 250 381 55 17250 383 19 19.2 19.2 19.2 4.3 4.5 4.4 250 368 39 21.3 20.8 21.05 6.6 6.3 6.45 250 306 56 250 367 21 20 19.8 19.9 4.5 4.8 4.65 250 408 25 19.3 19 19.15 4.7 5.2 4.95 250 358 61 22250 390 16 17.7 17.9 17.8 3.9 4 3.95 250 336 28 18.9 19 18.95 6.3 6.1 6.2 250 406 54 250 393 12 17.3 17.4 17.35 3.8 3.7 3.75 250 392 26 19.7 18.8 19.25 4.4 4.9 4.65 250 104 61 250 368 22 21.1 21.2 21.15 3.8 3.8 3.8 250 365 39 23 23 23 5.8 6 5.9 250 393 48 22

F1 growth measurements 

7/27/09 

8/3/200

9 

8/11/200

9 

SEX

DOSE ID

BW

tarsus1

tarsus2

avgtarsus

culmen1

culmen2

avgculmen SEX

DOSE ID BW

tarsus1

tarsus2

avgtarsus

culmen1

culmen2

avgculmen SEX

DOSE ID BW

tarsus1

tarsus2

F 0 

20

4 72 33 33 33 6.6 6.9 6.75 F 0 

20

7 96 33.1 33.1 33.1 7.2 7.4 7.3 F 0 

20

9 

10

5 32.8 33 

F 0 

23

0 83 34.2 34.1 34.15 6.3 6.4 6.35 F 0 

20

9 88 32.8 32.7 32.75 7.5 7.9 7.7 F 0 

21

1 

12

0 35.1 34.9 

F 0 

24

8 83 33.2 33.3 33.25 6.5 6.8 6.65 F 0 

21

0 

10

3 34.5 34.5 34.5 8 7.9 7.95 F 0 

21

7 

11

5 33.5 33.6 

F 0 

25

0 86 33 33.2 33.1 6.5 6.4 6.45 F 0 

21

1 

11

4 34.7 34.8 34.75 7.6 7.7 7.65 F 0 

21

9 

12

7 33.3 33.6 

F 0 

27

2 75 31.5 31.8 31.65 6.5 6.8 6.65 F 0 

21

5 87 32 32 32 7.3 7.1 7.2 F 0 

22

6 

11

9 34.9 34.9 

F 0 

27

5 79 32 31.7 31.85 7.1 6.9 7 F 0 

21

7 88 33.8 33.7 33.75 7.3 7.4 7.35 F 0 

23

4 

11

8 32.7 32.8 

F 0 

27

6 77 33.1 33 33.05 7.4 7.3 7.35 F 0 

21

9 

10

7 33.9 34 33.95 7.6 7.8 7.7 F 0 

23

5 

11

5 33 33 

F 0 

29

2 78 33 33 33 6.8 6.9 6.85 F 0 

22

6 

11

0 34.9 34.8 34.85 7.1 7.3 7.2 F 0 

23

7 

10

1 33.5 33.5 
B-1 

F 0 

29

7 77 32.4 32.4 32.4 6.3 6.6 6.45 F 0 

24

2 

10

7 32.5 32.6 32.55 7.6 7.6 7.6 F 0 

24

2 

11

7 32.4 32.5 

F 0 

29

9 72 31.1 31 31.05 7.6 7.4 7.5 F 0 

29

9 73 31.1 31.1 31.1 8.3 8.4 8.35 F 0 

24

5 

10

2 34 34.1 

F 30 

27

1 89 34.1 34.1 34.1 6.9 7 6.95 F 30 

20

6 89 31.3 31.4 31.35 8.8 8.7 8.75 F 30 

20

2 

11

3 33.5 33.3 

F 30 

27

3 64 30.6 30.6 30.6 6.4 6.2 6.3 F 30 

21

1 88 33.9 33.5 33.7 7.8 7.8 7.8 F 30 

21

2 

10

3 32.8 32.8 

F 30 

27

6 75 31.7 32.2 31.95 6.9 6.9 6.9 F 30 

21

2 90 32.4 32.5 32.45 7.8 8 7.9 F 30 

22

0 

12

8 32.9 32.9 

F 30 

28

0 81 33.2 33.2 33.2 7.4 7.3 7.35 F 30 

22

5 93 32.8 32.8 32.8 7.3 7.5 7.4 F 30 

22

6 

11

3 33.8 33.6 

F 30 

28

6 69 32.9 32.9 32.9 7 7 7 F 30 

22

6 

10

1 33.6 33.7 33.65 7.4 7.3 7.35 F 30 

23

0 

10

6 33.9 34 

F 30 

28

8 75 34.1 34.1 34.1 7.1 6.9 7 F 30 

23

1 87 33.6 33.8 33.7 8.2 8.1 8.15 F 30 

23

2 

11

1 33.5 33.5 

F 30 

29

0 80 32.4 32.5 32.45 7.1 7.1 7.1 F 30 

23

5 

10

3 34.7 34.7 34.7 7.8 7.7 7.75 F 30 

24

2 

11

0 33.6 33.5 

F 30 

29

1 81 33.3 33.3 33.3 6.6 6.5 6.55 F 30 

24

2 89 33 33.4 33.2 7.3 7.3 7.3 F 30 

24

5 

10

9 33.4 33.4 

F 30 

36

4 74 31.8 31.8 31.8 6.8 6.9 6.85 F 30 

24

7 98 34.3 34.4 34.35 7.4 7.5 7.45 F 30 

25

1 

11

2 33.4 33.4 

F 30 

36

9 76 33.1 33 33.05 6.7 6.9 6.8 F 30 

25

3 93 32.9 32.7 32.8 7.4 7.4 7.4 F 30 

25

3 98 33.2 33.1 

F 60 

10

5 73 33.8 33.8 33.8 6.6 7 6.8 F 60 

10

2 

10

1 31.7 31.7 31.7 8.1 8 8.05 F 60 

10

2 

12

2 31.6 31.6 

F 60 

12

2 78 33 32.9 32.95 7.1 7.1 7.1 F 60 

11

0 

11

5 34.3 34.3 34.3 7.4 7.1 7.25 F 60 

11

2 

10

4 33.4 33.5 

F 60 

12

4 71 33.7 34 33.85 6.9 6.9 6.9 F 60 

11

5 

10

9 35.2 35.7 35.45 7.4 7.5 7.45 F 60 

11

9 

12

7 35.5 35.5 

F 60 

13

9 62 34.1 34.3 34.2 7.3 7.5 7.4 F 60 

17

0 87 32.6 32.7 32.65 7.1 6.9 7 F 60 

16

2 

11

6 32.8 32.8 

F 60 

14

2 72 32.2 32 32.1 6.8 7 6.9 F 60 

17

4 88 33.1 33.1 33.1 7.9 7.3 7.6 F 60 

16

5 

10

5 32.2 32.2 

F 60 

14

3 66 31.3 31.1 31.2 7.4 7.3 7.35 F 60 

18

1 

11

3 33.1 33 33.05 7.8 8 7.9 F 60 

16

6 

12

4 33.4 33.3 

F 60 

15

3 70 32.8 32.9 32.85 6.7 6.9 6.8 F 60 

18

3 

10

1 33.6 33.6 33.6 6.9 6.8 6.85 F 60 

17

5 

12

1 33.3 33.6 

F 60 

15

4 50 29.2 29.4 29.3 5.9 5.8 5.85 F 60 

18

4 96 33 32.9 32.95 7.4 7.1 7.25 F 60 

18

2 

12

4 34.1 34.1 

F 60 

15

8 72 33.5 33.4 33.45 7.1 7.2 7.15 F 60 

18

5 97 33.6 33.5 33.55 7.5 7.2 7.35 F 60 

18

5 

11

5 33.8 33.8 

F 60 

19

5 55 30.2 30.2 30.2 7 6.9 6.95 F 60 

18

9 94 32.9 32.9 32.9 7.6 7.8 7.7 F 60 

18

6 

12

7 33.9 33.9 

F 125 

12

1 77 32.7 32.8 32.75 7.2 6.9 7.05 F 125 

10

1 

10

6 32.9 32.9 32.9 7.1 7.2 7.15 F 125 

10

1 

13

3 33.4 33.4 

F 125 

12

5 77 34.3 34.4 34.35 6.5 6.8 6.65 F 125 

10

3 

11

2 34.2 34.2 34.2 7.5 7.1 7.3 F 125 

10

3 

12

9 34 34.4 

F 125 

13

8 85 34.1 33.9 34 7 7.2 7.1 F 125 

10

6 

11

5 34.6 34.8 34.7 7.4 7.8 7.6 F 125 

10

6 

12

2 34.6 34.7 

F 125 

13

9 82 35.6 35.6 35.6 7 7 7 F 125 

10

7 90 32.4 32.4 32.4 7.1 7 7.05 F 125 

10

7 

10

7 32.3 32.1 

F 125 

14

3 77 32.1 32.6 32.35 6.3 6.1 6.2 F 125 

11

0 85 32.1 32.1 32.1 7.8 7.5 7.65 F 125 

11

0 

10

7 32.1 32 

F 125 

15

0 54 28.6 28.6 28.6 5.9 5.7 5.8 F 125 

16

0 98 32.5 32.5 32.5 7 7.2 7.1 F 125 

16

0 

11

5 32.3 32.3 

F 125 

17

8 81 34 34 34 8 7.6 7.8 F 125 

16

3 97 33.7 33.3 33.5 7.3 7.2 7.25 F 125 

16

3 

11

3 33.3 33.5 

F 125 

21

5 90 33.3 33.4 33.35 6.8 6.6 6.7 F 125 

16

5 94 33.8 33.8 33.8 6.9 7 6.95 F 125 

16

5 

10

0 33.7 33.7 

F 125 

22

0 81 34.3 34.2 34.25 7.2 7.3 7.25 F 125 

16

7 98 34.1 34.1 34.1 6.4 6.4 6.4 F 125 

16

7 

12

1 33.7 33.7 

F 125 

22

6 88 33.6 33.6 33.6 7.7 7.4 7.55 F 125 

19

1 99 33.2 33.2 33.2 7.9 7.5 7.7 F 125 

19

1 

10

8 33.2 33 

F 250 

12

2 80 32.6 32.6 32.6 7 7.1 7.05 F 250 

10

7 99 34.1 34 34.05 7.1 7.4 7.25 F 250 

10

7 

10

2 34.2 34.2 

F 250 

12

5 90 33.1 33 33.05 7.2 7.1 7.15 F 250 

11

4 

10

6 33.9 33.9 33.9 7.5 7.4 7.45 F 250 

11

4 

11

1 33.3 33.4 
B-1 

F 250 

14

3 83 33.3 33.3 33.3 6.9 6.9 6.9 F 250 

11

5 

11

0 34.5 34.7 34.6 7.3 7.5 7.4 F 250 

11

5 

12

0 34.1 33.8 

F 250 

14

5 84 32 31.8 31.9 6.2 6.3 6.25 F 250 

11

9 

10

4 34.6 34.5 34.55 7.6 7.9 7.75 F 250 

11

9 

10

9 34.4 34.4 

F 250 

14

7 89 33.1 33 33.05 6.7 6.5 6.6 F 250 

15

2 89 33.6 33.5 33.55 7.1 7.2 7.15 F 250 

15

2 

11

1 33.8 33.6 

F 250 

16

6 65 32.6 32.6 32.6 7.5 7.5 7.5 F 250 

15

4 97 34.7 34.7 34.7 7.4 7.7 7.55 F 250 

15

4 

10

5 34.5 34.6 

F 250 

17

0 68 31.8 31.8 31.8 6.6 6.3 6.45 F 250 

15

9 

10

3 34.1 34.2 34.15 7.9 7.9 7.9 F 250 

15

9 

12

2 34.1 34.2 

F 250 

17

1 86 32.4 32.4 32.4 7.1 7.2 7.15 F 250 

16

1 

10

9 34.7 34.8 34.75 7.4 7.5 7.45 F 250 

16

1 

12

7 34.6 34.4 

F 250 

17

6 91 32.4 32.3 32.35 6.7 6.9 6.8 F 250 

17

1 94 32.8 32.7 32.75 6.9 7.1 7 F 250 

17

1 

10

4 32.5 32.5 

F 250 

17

9 71 32.3 32.4 32.35 6.4 6.4 6.4 F 250 

19

3 98 33.2 33.2 33.2 6.9 7 6.95 F 250 

19

3 

12

3 32.9 32.9 

M 0 

20

1 80 32.4 32.6 32.5 7.4 7.4 7.4 M 0 

20

5 88 32.5 32.5 32.5 6.9 6.9 6.9 M 0 

21

6 93 33.3 33.4 

M 0 

23

2 75 30.9 30.7 30.8 6.9 6.5 6.7 M 0 

20

6 96 34.1 34.2 34.15 7.2 7.5 7.35 M 0 

21

8 97 33.2 33.2 

M 0 

24

9 85 34.2 34.2 34.2 6.7 6.7 6.7 M 0 

21

3 95 33.1 33 33.05 7 7 7 M 0 

22

1 

10

3 32.9 33 

M 0 

26

2 69 31.8 31.9 31.85 5.9 6.2 6.05 M 0 

21

4 

10

2 34.8 34.8 34.8 7.9 7.7 7.8 M 0 

22

2 89 33.5 33.5 

M 0 

26

5 87 32.5 33.3 32.9 7 7 7 M 0 

21

8 94 33.1 33.4 33.25 7.6 7.9 7.75 M 0 

22

4 93 33.7 33.6 

M 0 

26

7 60 29.5 29.4 29.45 5.9 5.9 5.9 M 0 

22

1 99 32.8 32.8 32.8 6.9 6.6 6.75 M 0 

22

7 98 32.6 32.6 

M 0 

27

1 74 32 32 32 6.6 6.7 6.65 M 0 

23

2 85 31.6 31.7 31.65 7.1 7.1 7.1 M 0 

22

9 91 33.3 33.4 

M 0 

27

9 69 32.2 32.1 32.15 7.3 7.5 7.4 M 0 

24

4 87 32.6 32.6 32.6 7.4 7 7.2 M 0 

23

6 80 33.7 33.6 

M 0 

28

0 80 32.8 32.9 32.85 7 7.3 7.15 M 0 

24

6 89 33.8 33.8 33.8 6.9 6.8 6.85 M 0 

24

0 95 31.5 31.5 

M 0 

29

1 76 31.8 32 31.9 7 6.9 6.95 M 0 

27

9 76 32.3 32.4 32.35 7.6 7.8 7.7 M 0 

28

0 

10

3 33.2 33.2 

M 30 

12

3 75 32 32 32 6.4 6.3 6.35 M 30 

10

9 96 33.3 33.3 33.3 7.1 7 7.05 M 30 

10

9 

10

0 33.6 33.5 

M 30 

12

9 78 32.2 32.2 32.2 6.5 6.3 6.4 M 30 

11

4 

10

6 32.8 33.3 33.05 6.4 6.6 6.5 M 30 

11

4 

11

0 32.9 32.9 

M 30 

14

1 75 32.7 32.6 32.65 6.5 6.4 6.45 M 30 

15

5 88 31.2 31.3 31.25 6.7 6.6 6.65 M 30 

15

5 99 31.2 31.1 

M 30 

14

5 75 32 32.1 32.05 7.1 7.1 7.1 M 30 

17

0 91 32.2 32.3 32.25 7.2 7.2 7.2 M 30 

17

0 

10

1 32.4 32.4 

M 30 

14

7 80 32.7 32.8 32.75 6.9 6.6 6.75 M 30 

18

2 91 32.5 32.6 32.55 6.8 6.4 6.6 M 30 

18

2 

10

1 32.5 32.3 

M 30 

21

2 74 32.1 32.2 32.15 6.9 7 6.95 M 30 

18

4 96 33 33.1 33.05 7.6 7.4 7.5 M 30 

18

4 

10

3 33.3 33.3 

M 30 

22

4 70 31.6 31.6 31.6 7 6.8 6.9 M 30 

18

5 

10

2 33 33 33 7.8 7.5 7.65 M 30 

18

5 

10

6 33.1 32.9 

M 30 

22

5 63 29.8 29.8 29.8 6.6 6.9 6.75 M 30 

18

8 96 32.4 32.3 32.35 6.7 6.5 6.6 M 30 

18

8 

10

0 32.4 33 

M 30 

22

8 81 33.1 33.1 33.1 7 6.9 6.95 M 30 

18

9 93 33.1 33.1 33.1 7.7 7.6 7.65 M 30 

18

9 97 32.9 33 

M 30 

25

5 79 32.3 32.2 32.25 6.1 6.4 6.25 M 30 

19

3 97 33.1 33.2 33.15 6.9 7.3 7.1 M 30 

19

3 

11

2 33.2 33 

M 60 

10

4 83 32.5 32.4 32.45 7.1 7.4 7.25 M 60 

10

6 

10

6 33 32.9 32.95 7.3 7.5 7.4 M 60 

10

5 

10

3 32.6 32.9 

M 60 

12

1 79 32.8 32.7 32.75 6.6 6.7 6.65 M 60 

10

9 92 33.4 33.6 33.5 6.8 7 6.9 M 60 

11

1 

10

4 33.3 33.3 

M 60 

12

7 75 33.3 33.3 33.3 7.2 6.9 7.05 M 60 

11

6 92 33.6 33.7 33.65 7.4 7 7.2 M 60 

11

2 95 32.7 32.7 

M 60 

14

9 88 31.7 31.8 31.75 6.7 6.8 6.75 M 60 

11

8 82 33 33 33 7.4 703 355.2 M 60 

11

8 

10

0 33 33.6 

M 60 

16

3 81 31.8 31.9 31.85 7.2 7 7.1 M 60 

12

0 93 32 32.3 32.15 6.7 6.6 6.65 M 60 

12

0 98 32.4 32.3 

M 60 

16

7 66 32.1 31.9 32 7 6.9 6.95 M 60 

13

1 93 32.6 32.7 32.65 6.9 6.9 6.9 M 60 

13

5 93 32.4 32.5 
B-1 

M 60 

16

8 84 33.4 33.3 33.35 7.3 7.3 7.3 M 60 

13

2 93 32.4 32.4 32.4 7 7 7 M 60 

13

8 95 32.4 32.3 

M 60 

16

9 83 33.3 33.2 33.25 7.1 7.2 7.15 M 60 

13

5 92 32.6 32.5 32.55 7.1 6.9 7 M 60 

14

0 

10

0 34.1 34 

M 60 

17

7 82 33 33.1 33.05 7.1 6.9 7 M 60 

14

0 93 34 33.9 33.95 7 7 7 M 60 

14

1 

10

2 32.4 32.4 

M 60 

19

5 82 31.7 31.6 31.65 6.6 6.7 6.65 M 60 

15

5 95 32.4 33 32.7 7 7.2 7.1 M 60 

15

8 99 32.7 32.6 

M 125 

12

1 71 31.7 31.7 31.7 7.5 7.5 7.5 M 125 

11

1 

10

3 33.7 33.6 33.65 7.6 7.7 7.65 M 125 

11

1 

10

8 33.9 33.7 

M 125 

12

8 67 32.8 32.9 32.85 7.4 7.3 7.35 M 125 

11

4 82 31 30.9 30.95 7.2 7.4 7.3 M 125 

11

4 89 30.4 30 

M 125 

13

2 74 33 32.8 32.9 6.8 7 6.9 M 125 

11

6 80 32 32.1 32.05 6.6 6.9 6.75 M 125 

11

6 95 32.1 32.2 

M 125 

13

4 54 31.3 31.4 31.35 6.5 6.5 6.5 M 125 

15

1 

10

0 33.3 33.4 33.35 8.1 7.8 7.95 M 125 

15

1 

10

7 33.3 33.3 

M 125 

13

6 79 33 33.1 33.05 6.7 6.6 6.65 M 125 

16

4 86 32 32.4 32.2 7.6 7.5 7.55 M 125 

16

4 

10

6 32.3 32.3 

M 125 

14

9 69 31.1 31.2 31.15 7 6.8 6.9 M 125 

16

7 98 33.3 33.3 33.3 7 7.1 7.05 M 125 

16

7 98 33.5 33.7 

M 125 

19

3 72 31.4 31.5 31.45 6.5 6.5 6.5 M 125 

17

6 

11

2 34.3 34.1 34.2 6.9 6.9 6.9 M 125 

17

6 

11

1 34.7 34.5 

M 125 

19

6 61 31.6 31.6 31.6 6.6 6.4 6.5 M 125 

17

8 98 33.2 33.2 33.2 6.9 7.1 7 M 125 

17

8 

10

0 32.9 33 

M 125 

21

3 65 31.4 31.4 31.4 6.8 7.1 6.95 M 125 

18

7 99 33.6 33.5 33.55 7.4 7.3 7.35 M 125 

18

7 

10

0 33.6 33.6 

M 125 

21

4 67 32.7 32.7 32.7 6.6 6.5 6.55 M 125 

19

0 95 33.4 33.4 33.4 7.4 7.4 7.4 M 125 

19

0 

10

1 33.6 33.4 

M 250 

26

7 79 34.2 34.3 34.25 7.2 6.8 7 M 250 

20

5 93 34.1 34 34.05 7.3 7.1 7.2 M 250 

20

5 

10

3 34.1 34.1 

M 250 

27

5 85 31.5 31.5 31.5 7.3 7.5 7.4 M 250 

21

5 89 32.9 32.6 32.75 7.1 7 7.05 M 250 

21

5 

10

8 33 33 

M 250 

27

9 69 32 32.1 32.05 6.5 6.6 6.55 M 250 

22

3 93 33.8 33.9 33.85 7.3 7.4 7.35 M 250 

22

3 97 33.4 33.3 

M 250 

28

5 73 32.9 33.3 33.1 7 6.7 6.85 M 250 

23

7 96 33.6 33.6 33.6 6.8 6.7 6.75 M 250 

23

7 

10

5 33.7 33.7 

M 250 

29

4 80 32.1 32.4 32.25 7.6 7.3 7.45 M 250 

24

4 90 32.9 33 32.95 6.7 7 6.85 M 250 

24

4 98 32.9 32.9 

M 250 

29

6 81 31.1 31.2 31.15 6.3 6.6 6.45 M 250 

24

6 96 34.2 34.2 34.2 8 7.8 7.9 M 250 

24

6 

10

4 34.1 34.4 

M 250 

29

7 74 32.3 32.5 32.4 6.7 6.5 6.6 M 250 

25

5 87 31.8 31.6 31.7 7.6 7.3 7.45 M 250 

25

5 94 32 32.2 

M 250 

32

2 76 30.8 30.8 30.8 7.2 7.3 7.25 M 250 

25

7 96 33.7 33.7 33.7 7.3 7.2 7.25 M 250 

25

7 

10

0 33.6 33.5 

M 250 

32

6 68 30.3 30.2 30.25 6.3 6 6.15 M 250 

28

2 92 32.6 32.5 32.55 7.1 7.3 7.2 M 250 

28

2 96 32.5 32.4 

M 250 

32

7 79 32.8 32.8 32.8 6.8 6.7 6.75 M 250 

32

5 94 33.2 32.8 33 7.5 7.3 7.4 M 250 

32

5 

10

2 32.7 32.7 

F1 bird weights 

 8/19 8/24 9/1 9/9 9/14 9/21 9/29 10/6 10/13 10/20 10/27 11/2 11/10 11/17 11/24 8/19 8/24 9/1 9/9 9/14 9/21 9/29 10/6 10/13 10/20 SEX DOSE ID BW BW BW BW BW BW BW BW BW BW BW BW BW BW BW SEX DOSE ID BW BW BW BW BW BW BW BW BW BW M 0 221 110 113 114 113 116 117 114 116 118 117 117 116 116 122 114 F 0 217 128 129 122 129 126 134 125 135 138 134 M 0 218 105 107 106 105 108 107 107 109 110 110 110 111 109 111 107 F 0 211 137 138 137 144 142 123 135 145 144 144 M 0 236 104 108 108 107 109 108 107 111 112 111 113 115 113 116 111 F 0 234 114 130 124 131 127 133 134 134 125 M 0 224 110 115 116 116 115 116 111 113 116 115 118 117 114 120 115 F 0 237 122 126 138 127 136 137 131 M 0 216 100 102 104 106 105 104 104 109 110 109 109 110 104 104 101 F 0 235 113 114 120 120 118 130 126 128 134 133 M 0 227 101 101 103 102 106 101 105 109 111 109 110 111 111 113 110 F 0 245 112 128 127 136 135 131 133 142 146 142 
B-1 

M 0 280 104 103 106 106 108 104 107 113 109 114 115 113 115 118 117 F 0 219 101 121 127 133 132 126 137 136 137 137 M 0 240 97 98 98 98 103 103 101 100 102 103 102 102 103 102 104 F 0 242 115 132 128 130 132 132 127 119 134 135 M 0 222 95 97 99 97 98 101 99 101 102 101 101 104 103 104 103 F 0 226 123 132 133 126 136 138 138 142 143 143 M 0 229 99 101 102 103 104 103 101 101 104 104 105 105 105 107 108 F 0 209 130 123 125 130 126 135 134 137 138 136 M 0 243 98 101 106 107 108 108 98 102 103 103 105 107 106 108 110 F 0 215 103 119 123 127 124 125 127 130 125 126 M 0 220 109 111 112 111 113 117 115 116 117 116 119 120 122 123 122 F 0 299 86 92 106 114 116 119 119 122 122 124 M 0 213 103 104 107 104 104 104 F 0 203 123 136 134 129 136 139 138 141 141 132 M 0 279 90 89 93 94 96 93 93 94 96 94 97 97 91 94 F 0 228 121 130 130 101 129 129 129 130 131 132 M 0 214 115 116 117 117 117 118 112 116 119 120 120 121 116 121 113 F 0 225 128 137 135 139 134 134 137 129 146 144 M 0 232 97 99 99 99 100 100 104 105 108 107 104 103 104 108 101 F 0 241 133 136 129 129 124 127 134 147 128 141 M 0 244 97 98 101 104 104 103 103 103 105 106 108 143 104 107 100 F 0 212 124 128 129 118 131 131 131 123 134 132 M 0 206 105 110 110 111 113 112 107 108 111 114 112 113 112 115 110 F 0 202 124 129 133 130 134 136 123 132 135 130 M 0 246 102 103 102 104 104 101 102 104 107 106 105 108 109 F 0 207 115 120 133 129 132 136 118 106 136 134 M 0 205 97 98 99 98 99 98 97 100 162 99 101 102 104 106 F 0 210 115 129 136 M 30 257 104 105 104 104 104 105 106 110 110 105 109 109 109 109 110 F 30 253 108 109 114 125 128 128 132 134 138 136 M 30 255 99 102 103 103 105 106 104 107 107 108 108 110 112 133 114 F 30 226 144 143 148 139 144 140 141 147 143 143 M 30 246 109 110 109 110 114 112 110 115 117 113 115 116 116 118 117 F 30 212 107 124 138 136 126 136 138 134 120 140 M 30 237 113 117 118 116 119 116 116 118 117 118 120 120 121 125 125 F 30 242 116 126 144 137 142 138 130 144 125 146 M 30 325 112 115 115 116 116 115 115 120 119 117 119 121 123 122 123 F 30 202 128 140 136 139 138 130 138 141 139 139 M 30 282 104 106 109 106 108 108 105 110 109 110 110 113 109 111 112 F 30 230 136 137 137 138 133 141 139 146 143 146 M 30 223 108 109 108 111 111 115 106 110 112 110 111 111 111 114 112 F 30 232 115 120 145 138 145 140 131 145 146 146 M 30 244 97 100 102 100 102 100 105 105 106 107 107 106 106 109 109 F 30 220 126 123 138 134 130 134 128 135 134 135 M 30 215 97 115 117 117 116 112 118 119 119 121 121 118 122 125 121 F 30 245 125 125 127 124 127 127 130 129 133 132 M 30 205 105 112 113 111 112 113 113 115 115 115 114 116 113 117 118 F 30 251 137 126 139 136 139 138 146 138 139 149 M 30 224 102 107 108 108 110 103 109 110 111 109 112 111 112 113 111 F 30 227 126 121 133 126 134 130 137 135 129 138 M 30 233 97 104 105 103 105 108 105 108 109 109 110 109 110 110 109 F 30 250 130 143 142 144 142 146 139 144 139 149 M 30 219 102 103 104 104 106 108 108 106 106 104 103 105 104 106 107 F 30 247 127 131 137 139 139 139 145 143 145 142 M 30 248 101 102 105 106 108 104 107 110 111 111 111 112 114 114 116 F 30 211 127 140 136 136 135 138 137 133 139 145 M 30 216 113 115 117 115 115 115 111 115 118 116 117 120 122 122 123 F 30 231 115 131 142 144 145 137 139 147 136 150 M 30 217 101 106 109 112 107 108 101 107 109 108 110 109 111 115 F 30 235 135 133 143 137 135 142 144 133 147 147 M 30 240 98 99 102 99 102 104 102 108 107 105 108 108 108 108 107 F 30 225 115 120 126 119 127 125 127 131 130 129 M 30 272 104 105 105 103 103 101 100 104 103 108 107 100 96 107 F 30 206 110 124 127 128 123 140 136 130 123 134 M 30 204 109 113 113 106 110 87 93 113 115 120 121 122 123 123 123 F 30 249 129 130 140 140 138 134 137 128 134 136 M 30 209 100 103 108 108 110 100 106 105 106 107 107 107 109 110 110 F 30 221 127 133 134 138 132 141 137 145 143 136 
B-1 

M 60 176 113 117 119 119 120 114 99 120 123 130 129 128 131 130 F 60 162 96 112 132 133 131 129 111 134 113 134 M 60 167 102 103 103 103 107 102 108 109 108 109 112 112 118 116 F 60 166 128 133 134 134 135 132 136 134 137 144 M 60 116 101 102 103 100 101 101 100 103 105 104 106 106 105 110 107 F 60 182 135 132 139 131 138 103 108 125 128 127 M 60 111 117 120 121 121 123 122 116 120 122 123 114 123 124 127 123 F 60 112 118 120 128 131 126 133 128 129 123 140 M 60 164 104 105 108 108 110 109 107 108 112 114 124 116 116 120 118 F 60 119 139 140 145 142 137 147 133 106 142 129 M 60 151 116 121 127 130 133 131 128 129 127 128 124 126 127 130 F 60 186 135 134 140 135 146 143 147 136 137 149 M 60 114 97 98 100 100 101 101 99 100 102 102 103 104 102 106 104 F 60 165 113 115 113 115 102 119 117 116 125 125 M 60 187 103 104 104 105 105 106 108 110 112 112 112 114 115 118 117 F 60 175 134 131 136 135 133 131 121 146 121 144 M 60 190 107 108 107 106 107 110 108 112 114 110 114 105 109 118 118 F 60 102 102 125 132 122 134 129 129 136 135 193 M 60 178 105 107 110 111 110 109 112 115 113 114 107 115 114 F 60 185 185 132 133 134 134 137 132 134 131 132 M 60 177 103 105 107 107 106 104 106 110 109 112 113 95 102 114 111 F 60 184 184 124 137 136 136 133 125 137 136 136 M 60 180 96 99 100 100 99 97 98 99 99 101 101 94 97 103 105 F 60 181 181 128 135 120 137 136 124 131 140 141 M 60 161 110 118 124 125 129 126 126 131 135 135 136 132 130 137 F 60 183 183 123 131 139 136 142 141 141 146 146 M 60 109 101 101 104 104 103 104 107 109 109 110 111 96 105 113 114 F 60 174 174 110 111 121 112 121 122 126 125 127 M 60 173 110 113 116 114 117 117 115 118 114 113 116 104 113 116 119 F 60 189 109 121 119 123 123 122 122 127 118 126 M 60 120 103 105 105 104 105 107 106 109 108 109 109 99 103 110 114 F 60 170 130 137 136 129 136 130 121 124 131 137 M 60 169 104 107 110 112 114 112 112 115 118 119 118 112 105 119 F 60 110 129 129 137 138 136 137 132 112 133 143 M 60 171 111 112 115 111 112 110 108 110 115 118 120 107 111 118 120 F 60 115 136 143 144 131 142 143 140 146 142 144 M 60 117 113 115 116 112 116 116 116 115 117 117 116 107 112 122 121 F 60 179 130 114 137 134 118 136 116 142 129 136 M 60 136 107 116 117 116 115 116 114 114 115 118 119 108 119 120 120 F 60 172 122 132 119 130 131 134 131 137 128 139 M 125 114 119 120 121 122 123 122 124 126 127 128 127 127 126 127 126 F 125 101 131 126 135 131 137 134 134 138 138 139 M 125 109 112 112 115 115 119 121 113 113 116 118 119 116 118 122 118 F 125 103 139 143 142 144 138 130 144 148 140 148 M 125 184 113 112 114 115 117 115 112 119 120 118 121 122 120 125 124 F 125 110 131 132 127 128 132 138 128 120 132 139 M 125 193 124 128 131 130 134 133 129 130 125 128 127 131 133 129 132 F 125 165 120 135 138 136 136 138 133 94 M 125 182 103 104 105 105 105 106 107 107 107 105 108 107 109 111 110 F 125 107 127 141 132 136 141 138 145 133 148 145 M 125 155 101 105 104 103 104 104 103 138 105 105 104 104 104 109 113 F 125 167 119 130 135 132 127 133 129 134 136 139 M 125 170 103 103 101 101 101 88 102 145 105 104 104 100 107 109 108 F 125 191 127 136 135 134 136 135 138 127 122 134 M 125 185 112 114 114 113 116 114 112 110 111 116 117 114 113 120 119 F 125 106 121 121 130 131 141 128 142 136 146 141 M 125 189 102 104 106 103 104 104 104 105 107 109 109 108 108 111 108 F 125 160 130 132 135 137 134 138 140 136 141 135 M 125 188 104 108 111 110 112 110 109 110 110 112 114 113 113 115 115 F 125 163 132 131 133 135 136 132 130 137 138 137 M 125 133 98 101 101 100 102 101 99 100 99 103 106 105 106 110 110 F 125 112 129 139 139 139 142 133 145 102 135 146 M 125 151 98 103 103 102 103 104 102 103 104 105 108 105 106 108 F 125 166 122 123 131 133 132 124 144 110 120 137 M 125 183 102 107 107 108 107 106 106 111 110 109 109 111 108 113 111 F 125 117 139 137 139 138 130 142 140 144 146 148 M 125 161 100 99 99 97 100 101 100 101 103 102 102 102 103 90 103 F 125 156 120 134 129 136 122 136 130 135 140 140 
B-1 

M 125 131 112 109 110 107 110 112 109 113 115 115 113 113 115 116 115 F 125 158 120 123 130 136 133 138 109 140 140 138 M 125 118 107 106 111 114 114 109 102 105 109 110 110 112 110 113 115 F 125 159 119 129 128 129 136 140 132 135 122 126 M 125 157 109 111 114 112 113 111 111 112 113 112 112 113 112 112 120 F 125 135 126 127 131 133 120 133 136 125 122 119 M 125 152 116 118 113 113 111 114 112 117 116 116 117 115 117 118 119 F 125 181 124 136 136 137 135 143 137 145 145 141 M 125 134 108 110 110 109 111 112 113 116 113 112 113 115 116 118 119 F 125 111 125 138 143 139 149 146 155 152 146 146 M 125 119 105 107 107 106 107 106 108 107 108 109 110 110 110 113 117 F 125 113 128 136 125 138 139 140 145 145 146 149 M 250 158 105 107 110 112 112 109 142 112 112 112 117 116 117 118 118 F 250 161 136 136 141 140 137 147 142 141 147 148 M 250 141 106 107 110 109 110 111 110 112 114 113 113 114 116 118 117 F 250 152 125 132 133 131 112 139 136 138 137 132 M 250 138 105 107 110 108 105 103 105 110 113 113 116 115 115 118 117 F 250 107 133 122 130 140 137 138 137 141 138 138 M 250 111 105 109 104 103 105 107 109 112 110 116 115 118 117 119 121 F 250 115 137 143 136 142 145 139 150 150 151 150 M 250 105 108 110 107 107 108 109 108 110 114 115 113 112 116 115 114 F 250 114 123 136 137 135 125 136 139 149 145 139 M 250 112 102 103 102 103 105 104 106 108 110 109 110 111 111 111 114 F 250 159 136 138 135 138 139 139 128 140 139 139 M 250 135 100 103 100 101 103 101 103 102 103 103 101 103 103 106 105 F 250 193 125 128 131 131 130 132 131 134 140 139 M 250 118 110 111 113 115 116 116 111 114 114 114 117 116 115 120 120 F 250 171 120 131 137 139 140 137 140 122 142 144 M 250 140 106 106 107 109 105 107 107 112 115 116 119 115 115 116 119 F 250 154 117 121 127 133 124 128 139 131 139 135 M 250 120 97 99 101 100 103 103 102 104 104 104 107 108 107 111 109 F 250 119 132 130 140 140 132 143 M 250 155 114 118 118 114 118 116 114 105 117 118 118 118 119 119 119 F 250 156 149 147 143 149 144 150 149 154 153 151 M 250 109 94 107 99 100 102 102 105 105 107 106 105 106 107 110 109 F 250 134 119 134 132 137 122 138 134 131 143 142 M 250 116 105 106 112 112 115 114 114 115 118 117 118 118 119 121 121 F 250 110 143 144 148 139 149 149 153 M 250 131 109 110 106 105 105 106 105 107 108 109 109 109 111 110 111 F 250 102 140 142 142 150 146 150 144 150 148 156 M 250 132 99 104 115 105 104 105 105 104 105 106 105 103 106 109 107 F 250 153 126 128 128 124 130 129 126 135 134 139 M 250 106 110 114 114 118 118 117 120 120 121 120 119 121 122 129 129 F 250 133 125 122 130 133 124 128 136 137 133 137 M 250 108 111 113 117 115 117 116 115 117 115 118 117 120 118 122 121 F 250 151 139 135 141 141 140 134 142 128 143 145 M 250 174 101 106 103 103 102 104 106 103 105 106 106 107 109 112 110 F 250 113 130 120 125 128 123 133 131 134 137 137 M 250 177 107 109 111 108 110 108 110 111 111 111 110 112 113 115 F 250 122 113 123 140 135 128 139 136 145 144 146 M 250 103 102 104 103 104 105 108 106 107 108 107 107 108 109 110 110 F 250 136 143 139 145 136 145 141 149 148 150 147 
F1 feed consumption 

 8/18 8/25 8/31 9/4 9/8 9/14 9/18 9/21 

DOSE SEX ID Initial Final Final Initial Final Initial Final Initial Final Initial Final Initial Final Initial Final Initial 

0 F 217 402 306 434 229 434 348 467 343 438 305 443 335 437 366 

0 F 211 387 292 434 296 449 344 451 329 440 292 444 365 433 381 

0 F 234 389 293 435 301 456 346 449 334 451 281 442 337 458 376 

0 F 237 375 309 428 268 445 344 452 332 439 267 442 309 451 381 

0 F 235 395 341 445 332 450 343 454 347 445 299 448 337 431 353 

0 F 245 300 347 425 321 452 367 457 357 448 304 450 331 425 355 

0 F 219 399 348 433 358 450 362 460 354 453 311 453 361 436 367 

0 F 242 400 331 419 300 446 354 465 368 458 298 437 310 434 341 

0 F 226 399 299 425 274 443 357 457 344 448 293 448 329 433 355 

0 F 209 369 308 438 298 443 351 445 329 440 250 467 341 434 351 

0 F 215 367 299 435 361 444 359 456 346 450 296 469 333 447 347 

0 F 299 381 367 424 341 433 379 451 365 448 326 435 344 437 368 

0 F 203 400 303 431 285 451 332 464 328 455 281 448 300 437 341 

0 F 228 391 295 434 295 437 345 444 362 460 349 431 298 421 352 

0 F 225 408 343 439 294 438 336 449 332 442 265 444 298 429 336 

0 F 241 399 302 447 299 451 356 464 358 450 343 432 346 446 370 

0 F 212 395 348 445 317 451 360 457 358 455 317 446 336 441 373 

0 F 202 399 328 430 309 435 348 453 339 447 293 458 321 444 356 

0 F 207 403 350 454 360 441 362 458 353 447 293 453 327 438 336 

0 F 210 390 329 447 343 433 388 455 

0 M 221 408 351 445 359 438 350 453 347 436 356 438 331 

0 M 218 386 334 423 329 448 348 454 327 445 343 442 314 

0 M 236 435 351 430 329 440 341 447 335 447 365 440 332 

0 M 224 421 331 421 325 453 352 453 337 447 340 439 297 

0 M 216 410 342 426 326 445 339 450 322 441 338 436 307 

0 M 227 398 332 425 353 452 356 456 319 442 338 454 326 

0 M 280 433 344 435 357 452 357 458 331 437 336 434 313 

0 M 240 431 346 437 365 424 348 456 358 442 351 455 344 

0 M 222 434 337 430 336 444 341 449 324 442 352 443 321 

0 M 229 421 354 439 349 443 346 455 337 440 354 452 337 

0 M 243 435 344 430 353 437 348 450 336 442 372 431 343 

0 M 220 425 329 432 311 435 314 440 294 438 320 443 283 

0 M 213 425 351 446 358 437 343 457 335 440 342 437 311 

0 M 279 400 343 451 373 432 345 447 331 443 354 442 309 

0 M 214 405 330 435 309 429 305 459 300 443 315 448 290 

0 M 232 420 314 438 360 443 353 440 327 448 354 447 312 

0 M 244 420 342 439 365 438 343 444 320 447 353 439 331 

0 M 206 417 318 440 336 409 302 446 306 427 318 434 321 

0 M 246 438 335 444 357 438 338 448 322 441 338 448 320 

0 M 205 410 357 441 356 447 357 443 327 444 361 450 328 

30 F 253 429 341 446 358 427 388 441 370 424 291 442 342 444 361 

30 F 226 411 313 434 254 453 337 447 337 429 275 428 322 439 352 

30 F 212 435 376 451 344 438 355 449 345 442 285 439 338 447 379 

30 F 242 419 340 460 354 437 361 437 344 436 290 450 334 458 369 

30 F 202 423 323 452 289 442 336 437 304 440 256 442 319 440 338 
B-1 

30 F 230 395 324 446 313 446 330 453 331 439 278 447 325 440 350 

30 F 232 412 341 443 348 455 350 450 349 454 300 443 317 451 373 

30 F 220 401 331 450 319 432 345 440 342 441 305 462 353 450 376 

30 F 245 418 330 460 343 438 366 456 360 455 302 446 348 461 392 

30 F 251 400 325 443 314 440 350 439 344 452 289 439 337 441 360 

30 F 250 396 311 443 307 434 320 439 292 446 265 439 308 438 342 

30 F 247 410 327 447 315 452 334 447 326 444 291 454 439 447 357 

30 F 211 398 323 427 317 436 349 444 350 447 304 420 321 431 360 

30 F 231 411 325 449 321 432 352 448 344 450 298 442 310 444 356 

30 F 235 405 361 438 327 434 335 448 340 445 299 450 328 446 359 

30 F 225 419 329 444 320 453 356 447 349 448 300 439 318 450 368 

30 F 206 403 348 441 321 442 353 446 344 449 322 441 340 439 374 

30 F 249 410 333 445 318 433 351 448 336 447 301 437 324 455 374 

30 F 221 407 312 449 342 442 359 450 346 457 315 461 340 446 366 

30 M 257 417 353 421 328 431 338 435 279 444 340 422 286 

30 M 255 415 361 437 359 434 351 445 344 450 370 441 314 

30 M 246 413 332 440 327 424 326 434 307 450 337 445 301 

30 M 237 402 353 449 358 431 339 440 331 445 364 438 321 

30 M 325 427 357 444 353 443 347 448 332 462 363 439 310 

30 M 282 425 348 436 336 439 331 441 323 449 353 412 300 

30 M 223 415 333 440 340 433 332 454 317 448 342 439 312 

30 M 244 413 358 444 365 440 355 437 330 446 366 449 345 

30 M 215 426 349 441 355 438 351 438 331 451 370 434 332 

30 M 205 423 355 435 338 439 344 446 307 447 357 429 292 

30 M 224 425 338 433 349 434 350 445 335 440 351 416 323 

30 M 233 431 348 423 334 436 329 438 310 446 340 442 305 

30 M 219 409 356 442 354 439 348 439 322 435 338 442 306 

30 M 248 421 341 437 346 437 342 438 300 456 350 427 328 

30 M 216 432 352 431 344 438 348 445 327 453 367 433 334 

30 M 217 421 342 438 357 438 341 440 315 459 376 458 344 

30 M 240 424 357 422 346 421 338 444 347 443 362 437 323 

30 M 272 415 368 446 358 452 359 445 342 443 371 437 339 

30 M 204 433 361 445 341 451 356 450 347 456 359 460 355 

30 M 209 431 318 439 323 424 295 457 305 453 334 444 316 

60 F 162 424 395 443 354 443 352 445 345 440 299 448 336 431 364 

60 F 166 417 326 428 303 446 367 439 346 451 316 448 348 443 385 

60 F 182 426 321 444 307 440 352 449 345 457 318 451 360 454 429 

60 F 112 417 332 436 336 445 363 445 360 454 327 442 342 449 385 

60 F 119 407 270 445 255 457 323 453 319 452 268 451 317 435 340 

60 F 186 426 320 461 312 451 348 439 351 453 322 453 346 457 381 

60 F 165 412 312 441 307 456 347 444 321 458 322 453 327 445 353 

60 F 175 424 322 449 309 452 352 448 341 452 305 447 324 440 362 

60 F 102 418 311 434 293 436 339 435 317 451 283 429 278 442 355 

60 F 185 418 321 443 314 452 334 440 322 442 276 445 333 437 347 

60 F 184 425 308 448 309 434 346 453 330 439 277 442 305 438 341 

60 F 181 418 329 437 332 440 359 455 359 461 331 438 302 426 344 

60 F 183 416 353 452 346 443 355 454 358 447 302 440 299 434 354 

60 F 174 416 317 449 320 433 346 453 334 452 319 444 344 444 362 
B-1 

60 F 189 430 322 450 326 432 356 437 342 449 296 462 338 436 352 

60 F 170 411 323 443 315 442 358 437 329 445 286 446 318 435 351 

60 F 110 429 317 438 297 433 350 437 335 441 287 443 336 445 365 

60 F 115 428 311 452 306 451 351 450 363 453 301 441 310 448 361 

60 F 179 423 303 448 320 444 356 450 335 446 333 438 327 445 367 

60 F 172 422 309 449 297 442 359 447 342 445 281 437 277 424 346 

60 M 176 424 376 424 334 439 340 441 305 438 324 429 295 

60 M 167 423 361 449 352 448 342 452 305 453 336 448 317 

60 M 116 405 348 430 349 456 378 454 355 451 383 447 347 

60 M 171 409 318 461 334 443 331 436 312 458 356 440 293 

60 M 164 421 353 444 340 454 317 444 299 464 335 438 285 

60 M 151 412 328 436 317 429 308 436 291 456 346 429 315 

60 M 114 420 351 429 345 442 357 446 345 454 381 423 328 

60 M 187 401 342 435 336 438 337 443 355 456 355 438 319 

60 M 190 413 343 432 350 433 355 445 322 458 353 438 310 

60 M 178 416 358 435 355 423 355 438 330 442 345 435 316 

60 M 177 431 352 442 348 460 368 452 333 444 358 454 338 

60 M 180 411 355 443 364 454 368 452 340 450 362 443 337 

60 M 161 413 336 440 350 457 327 444 294 453 315 433 281 

60 M 109 406 353 431 334 451 338 435 313 444 342 436 295 

60 M 173 413 358 434 337 444 344 444 320 440 337 428 302 

60 M 120 423 337 433 325 439 326 440 284 451 311 432 334 

60 M 169 419 355 437 329 433 341 445 336 453 366 433 322 

60 M 171 412 318 461 334 443 319 457 312 458 356 440 293 

60 M 117 423 349 439 349 450 352 456 352 451 355 424 279 

60 M 136 416 350 439 342 441 324 447 319 449 354 430 330 

125 F 101 416 302 428 289 423 338 448 346 442 294 454 323 445 334 

125 F 103 421 344 427 290 429 355 437 345 445 312 462 346 453 364 

125 F 110 409 315 436 307 423 353 438 339 444 308 451 346 448 372 

125 F 165 403 328 449 331 448 350 447 343 451 323 436 318 459 381 

125 F 107 418 338 450 303 432 327 441 314 448 271 449 316 448 349 

125 F 167 400 306 439 304 446 336 445 332 439 289 436 319 456 368 

125 F 191 440 331 456 324 436 339 438 332 452 298 444 332 457 365 

125 F 106 431 355 446 369 441 391 448 371 444 320 431 318 450 370 

125 F 160 439 307 448 340 440 327 442 330 447 284 448 324 459 373 

125 F 163 429 217 444 295 423 343 441 325 450 296 420 303 454 360 

125 F 112 407 333 447 302 422 346 445 237 429 277 447 330 452 366 

125 F 166 415 374 448 338 441 339 457 340 450 310 435 316 446 369 

125 F 117 428 319 450 318 439 342 441 335 440 296 441 330 452 371 

125 F 156 432 345 452 330 443 350 435 329 442 323 445 327 461 370 

125 F 158 428 345 445 358 439 371 448 358 450 318 435 339 457 393 

125 F 159 427 337 448 351 448 387 436 359 444 317 430 316 448 372 

125 F 135 415 365 445 326 437 353 437 337 447 326 435 337 453 379 

125 F 181 431 311 452 309 438 341 446 335 453 313 423 302 441 366 

125 F 111 433 321 448 313 452 348 449 327 443 277 443 303 448 354 

125 F 113 425 342 456 323 429 355 448 341 449 289 443 323 460 370 

125 M 114 418 342 454 345 441 341 440 327 452 331 438 292 

125 M 109 410 331 458 367 445 354 420 278 446 347 437 306 
B-1 

125 M 184 410 348 438 349 431 346 402 313 442 354 427 321 

125 M 193 401 309 428 311 441 340 413 291 446 330 437 284 

125 M 182 419 357 437 344 438 347 410 293 459 349 446 322 

125 M 155 405 353 441 354 431 349 421 321 451 367 435 330 

125 M 170 424 368 451 368 425 367 432 326 455 361 453 352 

125 M 185 411 324 443 330 410 317 410 294 446 322 428 313 

125 M 189 432 362 444 352 437 352 409 311 453 377 445 341 

125 M 188 434 371 450 350 438 344 414 308 449 357 431 321 

125 M 133 427 364 439 354 443 366 413 311 461 361 438 331 

125 M 151 415 344 440 329 429 335 411 297 438 326 435 307 

125 M 183 412 347 436 322 448 383 440 307 442 327 427 282 

125 M 161 407 344 439 349 435 351 434 316 437 324 434 283 

125 M 131 403 327 442 358 431 338 435 311 441 325 424 290 

125 M 118 413 324 436 356 432 345 444 322 436 336 435 326 

125 M 157 410 347 444 332 443 321 413 308 452 339 437 285 

125 M 152 405 330 438 329 439 345 424 316 446 350 434 280 

125 M 134 423 329 424 313 432 320 424 284 437 319 432 290 

125 M 119 406 338 433 356 429 342 427 317 433 350 447 354 

250 F 161 413 321 433 299 434 351 439 348 453 322 457 345 438 369 

250 F 152 418 319 428 295 450 327 445 329 448 315 450 342 448 369 

250 F 107 406 329 444 298 426 341 442 325 448 278 441 309 454 339 

250 F 115 408 338 448 306 423 344 437 337 446 288 445 337 453 378 

250 F 114 431 335 448 330 440 355 454 347 446 326 449 334 465 392 

250 F 159 401 278 453 298 432 332 440 322 449 275 447 329 457 362 

250 F 156 418 281 450 243 459 305 430 276 450 238 445 253 443 324 

250 F 134 427 334 454 321 441 339 448 330 446 297 453 328 452 359 

250 F 110 425 299 438 247 446 316 443 304 450 258 429 344 447 346 

250 F 102 418 310 428 291 435 339 439 298 446 262 428 282 450 337 

250 F 153 405 310 445 288 437 329 439 301 455 273 432 289 446 340 

250 F 133 413 317 451 332 435 343 441 327 451 285 431 315 461 363 

250 F 151 422 300 440 271 443 307 432 300 452 249 430 283 438 331 

250 F 113 413 302 432 295 425 353 437 334 445 288 434 322 452 383 

250 F 122 423 335 437 333 440 347 444 330 444 306 429 318 461 376 

250 F 136 431 312 439 281 446 322 438 306 453 286 446 295 463 348 

250 F 193 414 291 448 318 447 278 443 318 458 371 

250 F 171 428 330 437 353 442 297 444 330 460 376 

250 F 154 433 336 441 365 450 329 444 361 466 398 

250 F 119 326 329 438 304 432 339 448 330 453 316 443 327 450 358 

250 M 158 422 352 450 361 436 346 441 316 447 359 459 335 

250 M 141 416 344 442 358 438 351 439 336 452 363 415 305 

250 M 138 426 345 430 334 446 351 436 324 452 360 440 351 

250 M 111 422 362 443 364 439 361 449 341 450 358 438 323 

250 M 105 420 339 438 331 430 331 431 305 451 339 441 303 

250 M 112 422 334 450 336 451 351 449 319 444 341 450 312 

250 M 135 424 356 453 366 451 368 458 346 454 363 433 325 

250 M 118 411 330 424 308 431 313 437 298 445 335 428 299 

250 M 140 430 345 449 352 453 344 451 293 453 320 437 321 

250 M 120 416 365 444 368 440 341 429 312 461 350 432 333 
B-1 

250 M 155 420 304 442 335 422 330 445 354 458 364 450 336 

250 M 109 411 345 442 365 434 352 442 334 447 354 426 304 

250 M 116 427 348 426 340 440 346 444 325 452 367 431 311 

250 M 131 424 331 427 345 439 351 446 341 461 373 440 331 

250 M 132 413 348 449 361 444 357 454 334 461 369 434 324 

250 M 106 405 352 437 343 437 342 435 306 453 342 436 314 

250 M 108 408 326 438 292 448 318 437 286 449 318 429 284 

250 M 174 409 352 435 316 438 330 420 304 442 335 423 290 

250 M 177 421 331 432 328 449 347 435 332 445 344 441 301 

250 M 103 412 357 436 348 449 365 448 336 450 363 447 322 
F1 feed consumption 

 9/28 10/2 10/4 10/9 

10/1

2 

10/1

6 

10/1

9 

10/2

3 

10/2

6 

10/3

0 11/2 11/4 11/9 

11/1

3 

11/1

6 

DOSE ID Final 

Initial 

Final 

Initial 

Final 

Initial 

Final 

Initial 

Final 

Initial 

Final 

Initial 

Final 

Initial 

Final 

Initial 

Final 

Initial 

Final 

Initial 

Final 

Initial 

Final 

Initial 

Final 

Initial 

Final 

Initial 

Final 

0 

221/217 451 271 452 323 428 254 420 261 409 282 450 321 430 272 446 310 445 255 455 329 436 300 419 297 431 246 438 270 445 

0 

218/211 451 294 449 328 427 253 442 267 424 292 433 274 435 268 438 282 432 261 461 323 441 309 440 277 438 281 446 278 435 

0 236 435 368 498 402 433 374 429 356 406 355 453 397 429 359 441 384 416 356 465 409 443 394 442 376 452 392 448 401 448 0 

224/237 463 235 454 323 440 247 439 257 400 251 449 303 443 257 429 288 429 259 463 322 435 283 451 317 435 242 438 235 433 

0 216 434 334 457 378 429 350 441 346 434 364 444 375 438 362 444 367 413 343 462 386 442 375 459 367 451 366 449 372 451 0 

227/245 450 279 461 331 437 252 450 260 412 272 440 314 437 248 458 307 429 255 453 319 439 357 447 369 451 379 434 365 432 

0 

280/215 427 259 459 321 422 244 454 291 411 270 449 303 448 270 434 284 440 264 449 330 435 295 447 261 437 259 430 263 440 

0 

240/242 463 280 450 342 439 287 451 258 431 293 456 310 451 284 436 303 443 273 453 314 427 269 439 284 445 283 428 280 435 

0 

222/226 452 279 459 319 422 247 453 274 413 238 423 262 451 279 425 266 441 244 450 317 444 296 451 281 456 391 433 302 434 

0 

229/209 465 267 466 313 426 362 427 257 422 256 446 267 446 242 434 279 447 235 462 313 430 262 438 280 448 250 421 254 434 

0 243 439 388 464 418 418 246 446 352 404 358 450 393 436 365 424 381 447 386 458 410 433 387 448 390 437 378 437 374 421 0 

220/234 457 262 456 292 433 260 436 281 436 279 439 306 444 278 432 287 437 255 460 339 438 295 453 268 445 254 428 250 428 

0 

279/228 440 257 461 304 457 264 456 257 410 255 439 276 447 281 430 284 441 254 454 321 443 296 440 305 442 242 424 244 427 

0 

214/225 459 273 462 321 450 264 456 267 421 246 435 274 440 250 430 253 447 248 442 290 447 290 448 308 446 260 434 262 431 

0 

232/241 441 285 452 316 436 235 451 284 427 291 452 308 456 287 453 307 446 285 444 319 430 270 432 292 447 273 435 274 437 

0 

244/212 457 288 461 353 453 289 453 293 428 308 450 331 450 295 460 338 440 271 453 346 449 320 447 313 441 279 436 280 430 

0 

206/202 460 297 465 371 448 287 441 257 425 283 439 294 438 255 437 376 427 260 439 311 442 292 430 288 437 249 427 244 427 

0 

246/207 257 285 467 353 453 269 444 270 438 289 441 299 443 264 451 328 448 300 445 327 444 307 435 315 445 259 430 261 445 

0 205 452 365 464 389 402 342 437 323 428 371 444 384 428 359 441 372 436 358 460 385 454 379 440 358 439 356 447 357 432 0 235 422 318 462 373 428 328 438 340 388 319 442 367 437 335 441 347 445 322 465 370 434 348 447 336 444 307 450 345 448 0 299 431 352 460 388 388 319 441 345 386 313 446 366 437 338 466 378 426 347 436 376 436 365 454 359 449 344 448 348 450 0 219 436 325 455 383 441 344 439 306 386 305 436 351 430 330 439 349 401 396 456 365 440 348 438 341 460 348 454 306 432 0 203 454 323 458 347 421 276 446 323 389 309 456 347 429 305 444 336 416 283 461 354 446 336 446 313 440 273 433 252 428 

30 257 437 337 449 355 429 340 448 342 395 329 439 363 428 361 434 334 440 369 443 366 552 348 435 349 414 340 430 30 

255/212 449 284 454 336 430 260 434 319 382 279 438 331 454 337 426 368 435 311 444 359 418 321 422 280 433 292 415 297 430 

30 246 447 354 462 368 442 347 465 346 393 339 436 371 452 355 433 362 429 333 431 361 412 357 431 350 433 358 424 349 433 30 

237/242 462 288 451 312 429 262 442 294 402 292 436 308 445 297 434 318 438 266 440 318 426 303 441 281 439 389 416 392 431 

30 305 457 360 460 395 423 351 452 363 403 343 439 386 414 345 445 387 433 346 440 383 419 365 428 360 430 369 432 371 440 30 282 439 358 459 390 429 351 434 342 404 345 431 370 408 327 453 406 450 356 441 394 421 357 426 357 431 365 429 362 435 30 

223/232 463 296 457 312 435 250 448 257 411 269 435 294 444 253 420 286 441 254 444 308 426 297 438 267 434 374 452 370 433 

30 

244/220 450 288 441 316 439 276 443 281 401 289 442 386 425 301 436 304 439 270 443 312 429 300 421 278 441 291 417 292 414 

30 

215/245 452 286 452 310 449 269 443 280 423 288 447 315 441 276 442 310 430 271 428 299 423 291 451 301 435 279 419 280 420 
B-1 

30 

205/251 453 308 457 302 429 254 437 274 415 275 436 265 449 260 432 319 437 239 434 287 428 287 449 273 442 269 421 272 421 

30 224 343 459 371 426 337 437 386 425 260 427 377 450 356 440 392 426 383 432 339 436 367 427 367 425 30 

224/227 460 398 430 363 398 356 440 

30 

233/250 258 279 458 279 444 245 433 253 405 252 438 293 449 266 432 264 441 264 431 257 424 243 445 272 430 275 414 270 439 

30 

219/247 454 278 448 306 423 263 438 255 408 280 441 313 443 276 451 303 443 259 414 289 430 291 440 267 431 262 426 261 419 

30 

248/211 458 305 454 315 433 264 443 280 428 288 441 311 448 290 440 307 424 253 438 306 430 295 441 300 440 264 422 271 436 

30 216 437 369 460 401 428 366 429 355 440 280 435 379 441 367 422 362 441 356 429 373 426 369 421 345 428 30 

217/235 454 315 456 327 436 274 452 299 421 279 431 284 448 299 431 298 433 259 439 311 412 277 451 317 450 355 439 350 412 

30 240 458 378 458 365 440 368 452 362 432 345 426 379 417 348 435 374 439 351 440 386 429 377 447 363 433 351 427 360 422 30 

272/230 455 302 451 294 435 241 442 281 410 291 433 270 439 246 427 280 439 275 467 361 444 348 460 396 458 280 435 279 428 

30 204 265 368 466 385 459 385 465 373 458 341 433 356 438 337 428 360 445 335 439 375 431 374 439 353 420 335 421 330 433 30 

209/221 454 275 462 321 439 254 452 283 433 284 430 293 437 274 421 290 438 286 441 306 430 296 458 315 447 243 435 235 440 

30 253 456 353 454 367 430 325 455 339 413 331 437 341 443 331 415 324 329 303 432 345 436 339 415 293 449 320 460 322 427 30 226 429 261 446 351 439 303 448 322 427 308 438 329 433 302 435 265 434 294 445 330 417 304 411 247 428 254 450 250 430 30 202 450 314 457 350 432 275 445 329 403 267 440 334 438 246 418 328 441 243 452 351 424 323 423 259 440 298 454 300 433 30 231 454 438 454 338 436 319 463 340 409 316 442 344 424 304 421 321 418 278 437 351 419 334 419 302 441 294 456 296 441 30 225 453 340 460 345 432 325 421 314 408 327 436 351 430 322 418 325 428 316 434 348 425 343 432 324 424 316 448 317 440 30 206 453 357 452 376 435 335 451 345 395 269 434 366 434 342 416 271 446 336 441 345 421 332 427 326 429 337 440 335 438 30 249 455 354 446 383 444 357 449 353 405 351 447 367 444 362 430 385 429 390 450 404 436 364 437 335 454 327 442 330 429 30 227 457 281 442 296 427 341 454 281 428 343 426 321 437 354 414 297 412 267 426 278 423 375 435 60 

176/162 436 274 448 340 428 376 444 348 411 335 424 351 446 340 447 353 437 313 453 368 445 372 453 331 453 318 436 315 421 

60 167 464 376 440 375 452 355 452 357 420 357 440 374 447 345 433 368 441 338 448 378 432 362 451 349 426 322 433 330 429 60 

116/182 437 322 464 345 436 292 461 326 411 302 433 337 442 303 438 361 429 304 443 362 418 339 455 326 437 311 428 310 428 

60 

111/112 447 189 450 318 429 258 451 302 401 260 435 313 432 249 425 298 424 263 442 314 433 297 449 274 422 271 435 275 420 

60 

164/170 460 285 462 320 445 246 447 267 419 269 433 280 440 257 432 303 428 271 444 294 421 265 445 284 422 255 427 256 422 

60 

151/186 453 300 452 301 443 266 436 272 410 266 442 301 438 274 428 302 439 262 459 335 430 301 458 287 452 269 431 263 439 

60 

114/465 456 293 447 310 439 273 448 277 428 315 437 323 450 275 436 308 438 273 450 325 437 320 459 305 455 296 435 288 429 

60 

187/175 450 285 446 296 441 253 442 306 423 286 442 293 439 277 438 322 410 258 452 310 432 304 455 298 447 276 436 272 435 

60 173 437 360 460 392 448 382 450 382 401 361 439 385 443 350 434 382 435 339 451 399 436 386 441 365 436 359 433 362 440 60 

120/185 447 249 459 289 443 243 449 250 415 282 439 298 435 284 446 314 419 272 440 316 429 306 441 311 449 237 424 242 429 

60 169 441 365 462 386 433 357 427 342 420 361 444 374 432 343 416 343 440 328 436 357 431 363 436 354 421 341 431 345 427 60 

171/181 436 253 461 319 451 268 447 263 418 264 439 273 434 265 425 270 433 241 446 323 423 329 439 287 423 249 433 253 417 

60 

117/183 455 274 456 319 451 255 430 250 424 274 432 281 443 258 421 289 433 245 439 308 424 333 445 292 426 237 424 237 430 

60 

136/174 446 280 465 322 452 276 425 269 415 278 435 298 440 288 429 304 432 260 448 319 426 306 449 280 429 265 425 260 434 
B-1 

60 190 441 354 459 373 436 333 430 330 424 325 437 357 432 337 430 345 426 302 437 337 440 336 451 358 418 339 427 302 429 60 178 442 353 460 389 446 367 440 366 424 361 447 388 432 354 447 379 442 367 442 392 431 376 441 366 427 353 434 352 430 60 

177/110 459 317 467 357 455 314 443 

300

7 438 285 438 299 427 263 434 299 423 244 410 300 423 325 450 289 427 259 428 245 441 

60 

180/115 443 269 445 305 434 251 431 260 412 292 436 296 440 258 434 281 435 257 430 298 421 289 442 269 430 236 421 240 442 

60 161 442 330 467 373 447 347 436 328 441 356 434 359 435 346 432 355 445 344 443 367 426 359 451 361 420 327 435 371 430 60 

109/172 449 257 451 295 425 236 428 242 421 283 438 284 446 258 417 276 437 257 422 298 425 312 450 293 434 269 427 272 425 

60 166 449 357 452 389 437 349 432 344 421 353 445 367 435 328 452 377 437 338 437 366 421 359 453 359 432 333 441 330 420 60 119 442 336 444 341 435 283 438 322 425 323 440 356 434 324 455 360 435 311 449 353 425 333 456 320 447 311 435 315 432 60 102 449 328 452 353 419 304 453 326 430 341 436 343 432 317 444 347 430 283 434 347 421 331 454 320 444 299 435 300 419 60 184 449 342 462 354 433 319 448 309 429 334 433 344 426 315 438 349 443 312 442 336 420 317 443 308 439 289 434 288 425 60 189 465 350 462 370 441 332 447 324 453 347 442 353 430 324 451 360 443 311 446 350 424 332 458 338 440 295 423 290 402 60 179 456 360 464 376 445 355 431 346 422 343 437 344 425 301 423 338 434 303 446 351 436 337 447 325 454 311 436 302 420 

125 183 448 343 459 382 415 344 438 341 402 360 445 360 425 339 428 347 437 327 449 353 422 341 437 333 434 343 412 291 436 125 

161/103 451 259 450 315 437 246 422 266 415 291 426 293 441 267 422 291 428 261 437 288 435 287 439 264 424 240 417 340 420 

125 

131/110 437 262 456 317 440 260 430 262 422 277 434 270 434 244 418 268 427 236 423 276 432 278 443 265 443 253 422 249 433 

125 

118/165 447 319 450 354 428 288 427 350 430 360 450 370 446 342 444 376 442 346 437 375 439 368 437 355 419 343 410 249 428 

125 

157/107 459 286 468 334 442 259 447 267 423 253 439 276 437 235 436 294 424 240 450 287 434 279 444 251 422 328 433 341 427 

125 152 465 348 458 358 442 340 427 325 417 331 433 346 439 328 418 335 432 319 448 362 433 353 439 345 433 329 430 325 431 125 

134/191 450 314 446 376 438 259 419 277 407 276 452 325 439 280 422 270 431 261 445 312 432 297 448 292 453 248 408 331 435 

125 

119/106 458 290 450 331 442 266 423 273 413 293 446 315 438 271 420 273 426 262 440 300 442 303 450 266 440 272 416 245 436 

125 114 451 342 458 388 430 352 450 363 443 371 444 371 444 330 427 364 426 340 437 386 437 373 439 357 426 330 426 268 433 125 

109/156 464 307 461 314 426 272 448 275 408 261 449 316 435 271 420 286 430 271 443 310 429 288 442 275 429 268 429 333 427 

125 184 460 372 465 336 433 347 428 353 414 358 455 388 419 325 419 360 424 334 445 380 432 369 439 353 449 346 424 271 426 125 

193/159 457 288 454 323 434 239 435 293 423 340 457 341 459 330 437 337 441 257 449 308 433 273 432 274 422 240 414 350 422 

125 

182/135 462 295 457 328 436 254 419 278 421 321 444 331 443 305 436 305 441 254 440 305 417 254 425 271 421 235 422 242 439 

125 

155/181 462 305 456 302 422 269 440 276 413 303 437 310 439 256 434 310 432 269 443 312 440 311 431 284 436 279 420 236 427 

125 

170/111 453 292 446 313 441 287 425 278 424 286 456 308 449 272 425 293 418 252 451 306 433 297 433 282 423 263 413 277 435 

125 

185/113 457 280 446 292 442 267 434 259 422 269 449 274 445 236 445 287 425 243 441 300 430 259 429 258 427 255 412 260 421 

125 

189/160 455 296 447 309 449 287 422 268 408 279 451 320 444 254 413 292 435 256 442 313 426 265 417 274 422 280 424 261 435 

125 

188/158 446 285 461 317 448 246 423 243 417 269 445 299 451 254 423 252 432 253 446 306 444 308 433 278 434 276 421 275 434 

125 

133/112 462 336 461 338 444 276 427 290 419 391 453 312 443 265 413 251 431 251 446 324 448 308 442 236 434 262 415 242 437 

125 

151/166 459 289 449 300 431 277 420 282 423 363 450 383 430 336 427 338 427 343 446 380 433 368 444 360 433 417 436 429 436 

125 117 448 337 450 352 439 319 444 330 435 312 432 340 432 315 426 328 417 318 442 347 420 334 434 310 431 310 436 330 441 125 167 455 353 455 360 439 328 418 289 413 317 424 280 440 313 446 336 419 297 444 353 437 346 438 312 421 329 424 316 434 
B-1 

125 163 431 320 445 347 444 326 435 318 407 333 442 346 432 318 443 342 441 304 444 351 425 332 448 318 437 315 426 328 435 125 101 460 325 446 345 438 318 437 304 413 323 445 335 427 308 418 284 433 332 442 352 428 340 438 321 444 285 427 322 423 250 

135/156 444 237 452 285 435 230 427 261 395 242 437 264 441 242 446 262 429 262 447 261 435 246 441 270 440 256 414 260 413 

250 118 458 357 453 368 446 349 444 354 386 331 438 364 442 332 446 374 438 342 447 373 440 372 436 356 437 345 425 242 420 250 

140/110 459 334 452 338 429 334 420 337 402 326 435 344 432 334 433 348 433 344 449 380 440 374 441 354 423 333 429 342 422 

250 120 453 373 442 379 442 361 427 345 405 340 428 365 430 336 439 352 431 339 436 368 428 362 438 345 437 340 410 341 414 250 155 460 391 443 393 445 375 447 342 399 338 425 374 435 357 423 381 425 359 448 302 441 396 445 378 454 388 431 392 424 250 

109/133 456 291 450 388 433 230 434 239 392 267 436 305 443 275 435 298 440 265 440 309 439 296 447 287 430 264 401 270 429 

250 

116/151 461 277 448 319 440 249 426 260 393 263 445 296 445 289 439 294 423 280 433 255 438 291 448 251 469 234 415 241 426 

250 

131/113 457 292 454 317 436 261 429 269 393 261 435 291 432 253 424 297 425 276 425 287 427 292 447 257 411 286 429 269 416 

250 

132/122 462 279 450 297 441 255 437 273 394 267 440 283 443 261 445 280 439 259 446 293 450 285 440 266 425 256 417 265 427 

250 

106/136 453 251 446 294 431 230 425 256 415 282 445 299 436 296 434 305 438 273 448 303 442 303 449 283 431 253 421 252 431 

250 

108/102 452 255 452 279 432 234 429 262 400 246 443 242 434 243 435 290 435 245 443 263 437 258 413 268 434 251 410 254 427 

250 

174/171 448 298 442 289 438 266 420 267 394 261 435 290 443 276 434 302 438 258 444 293 446 302 426 294 431 252 407 250 435 

250 

177/154 452 280 448 320 433 253 426 263 399 228 447 313 444 283 435 314 425 269 443 294 437 290 420 291 425 256 422 251 441 

250 103 459 358 447 393 441 373 413 336 400 350 435 371 452 382 434 375 440 366 449 399 438 389 442 371 410 343 419 340 428 250 

158/161 458 307 460 399 439 278 442 279 407 284 435 296 440 275 423 280 431 261 426 292 439 287 421 273 428 273 413 268 434 

250 141 460 376 455 390 434 360 428 361 431 367 437 382 441 369 440 385 430 368 444 389 448 388 438 358 442 353 431 349 427 250 138 456 390 458 398 434 364 410 339 406 351 438 384 428 357 442 390 438 367 448 397 447 396 431 337 430 357 433 352 425 250 

111/115 450 298 445 305 433 257 425 287 407 306 444 303 446 288 430 305 436 264 453 332 425 305 445 263 445 271 420 270 424 

250 

105/115 450 257 445 300 434 234 416 267 427 259 448 262 446 260 427 295 427 253 426 266 423 251 435 259 415 264 418 265 433 

250 112 459 380 461 401 443 370 433 335 438 381 446 389 436 371 424 374 440 342 441 386 430 377 438 368 449 353 437 348 431 250 107 450 331 452 338 429 299 453 328 381 264 432 323 438 270 422 327 429 248 436 329 421 287 429 278 434 335 435 432 432 250 152 447 328 457 351 419 311 438 323 388 335 434 355 432 335 443 337 427 306 450 347 422 344 444 329 442 317 429 309 417 250 159 437 328 451 350 434 297 430 319 399 297 426 328 426 305 431 335 425 276 442 345 434 336 437 307 443 284 414 282 415 250 134 465 352 456 366 443 333 427 312 387 275 425 330 432 307 441 319 421 304 447 355 433 336 444 333 442 287 432 285 421 250 153 452 301 444 337 433 286 439 302 401 308 401 285 429 300 444 335 429 301 449 347 432 323 454 300 440 274 428 307 421 250 193 449 315 456 327 437 297 441 317 410 330 419 319 434 314 442 337 431 269 437 348 436 332 433 273 441 308 422 275 412 

F1 egg production

DOSE ID

8/

7

8/

8

8/

9

8/1

0

8/1

1

8/1

2

8/1

3

week 6

8/1

4

8/1

5

8/1

6

8/1

7

8/1

8

8/1

9

8/2

0

week 7

8/2

1

8/2

2

8/2

3

8/2

4

8/2

5

8/2

6

8/2

7

week 8

8/2

8

8/2

9

8/3

0

8/3

1

9/

1

9/

2

9/

3

week 9

9/

4

0

21

7 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

21

1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

23

4 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 
B-1 

0

23

7 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

23

5 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

24

5 1 1 1 1 1 1 1 1 

0

21

9 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

24

2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

22

6 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

20

9 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

21

5 1 1 1 1 1 1 1 1 1 

0

29

9 

0

20

3 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 

0

22

8 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

22

5 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

24

1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1

0

21

2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

20

2 1 1 1 1 1 1 1 1 1 1 1 

0

20

7 1 1 1 1 1 1 1 1 1 

0

21

0 

30

25

3 

30

22

6 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

21

2 1 1 1 1 1 1 1 

30

24

2 1 1 

30

20

2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

23

0 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

23

2 1 1 

30

22

0 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

24

5 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

25

1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

22

7 1 1 1 1 1 1 1 1 1 2 1 1 

30

25

0 1 1 1 1 1 1 1 1 1 1 1 1 

30

24

7 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 

30

21

1 1 1 1 1 1 1 1 1 1 1 1 

30

23

1 1 1 1 1 1 1 1 

30

23

5 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

22

5 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 
B-1 

30

20

6 1 1 1 1 

30

24

9 1 1 1 1 1 1 1 1 1 1 1 1 

30

22

1 1 1 1 1 1 1 1 1 1 1 1 

60

16

2 1 1 1 1 1 1 1 1 1 1 1 

60

16

6 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

18

2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

11

2 1 1 1 1 

60

11

9 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 

60

18

6 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

16

5 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

17

5 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 

60

10

2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

18

5 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

18

4 1 1 1 1 1 1 1 1 1 1 1 

60

18

1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

18

3 1 1 1 

60

17

4 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

18

9 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

17

0 1 1 1 1 1 1 1 1 1 1 

60

11

0 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 

60

11

5 1 1 1 1 1 1 1 1 1 1 1 1 

60

17

9 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

17

2 1 1 1 1 1 1 1 1 1 1 1 1 1 

125

10

1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

125

10

3 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

125

11

0 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

125

16

5 1 1 1 1 1 1 1 1 1 1 

125

10

7 1 1 1 1 1 1 1 1 1 1 1 1 

125

16

7 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 

125

19

1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

125

10

6 1 

125

16

0 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

125

16

3 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

125

11

2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 
B-1 

125

16

6 1 1 1 1 1 1 1 1 1 1 1 1 

125

11

7 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 

125

15

6 1 1 1 1 1 1 1 1 1 1 

125

15

8 1 1 1 1 

125

15

9 1 

125

13

5 1 1 1 1 1 1 1 1 1 1 1 1 1 

125

18

1 1 1 1 1 1 

125

11

1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

125

11

3 1 1 1 1 1 1 1 1 1 1 1 1 2 1 

250

16

1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

250

15

2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

250

10

7 1 1 1 1 1 1 1 1 1 

250

11

5 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

250

11

4 1 1 1 1 1 1 1 1 1 

250

15

9 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

250

19

3 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

250

17

1 1 1 1 1 1 1 1 1 

250

15

4 1 1 1 1 1 1 1 1 1 

250

11

9 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

250

15

6 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

250

13

4 1 1 1 1 1 1 1 1 1 1 1 

250

11

0 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

250

10

2 1 1 1 1 1 1 1 1 1 1 1 

250

15

3 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 

250

13

3 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

250

15

1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

250

11

3 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

250

12

2 1 1 1 1 1 

250

13

6 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 

F1 egg production

DOSE ID

9/

11 

9/

12 

9/

13 

9/

14 

9/

15 

9/

16 

9/

17 

week 11 

9/

18 

9/

19 

9/

20 

9/

21 

9/

22 

9/

23 

9/

24 

week 12 

9/

25 

9/

26 

9/

27 

9/

28 

9/

29 

9/

30 

10

/1 

week 13 

10

/2 

10

/3 

10

/4 

10

/5 

10

/6 

10

/7 

10

/8 

week 14 

10

/9 

##

0

217 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

211 1 1 1 1 1 1 1 1 1 2 1 1 1 
B-1 

0

234 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

237 1 1 1 1 1 1 1 1 2 1 1 1 1 2 1 1 1 1 1 1 1 1 

0

235 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

245 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

219 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

242 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

226 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 

0

209 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

215 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

299 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

203 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

228 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

225 1 1 1 1 1 1 1 1 2 1 1 1 1 2 1 1 1 1 1 

0

241 1 1 1 1 1 1 1 1 1 1 

0

212 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

202 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

207 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 

0

210

30

253 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

226 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

212 1 1 1 1 1 1 1 1 1 1 1 2 2 1 1 1 1 1 

30

242 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

202 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

230 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

232 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

220 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

245 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

251 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

227 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 

30

250 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 
B-1 

30

247 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

211 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

231 2 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 

30

235 2 1 1 1 1 2 1 1 1 1 1 1 1 1 1 

30

225 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

206 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

249 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

221 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

162 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

166 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 

60

182 1 1 1 1 1 1 

60

112 1 1 1 1 1 1 1 1 1 1 1 1 

60

119 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

186 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

165 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

175 1 1 1 1 1 1 1 1 1 1 1 2 

60

102 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

185 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

184 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

181 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

183 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

174 1 1 1 1 2 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 

60

189 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 

60

170 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

110 1 1 1 1 1 1 1 1 2 1 1 2 1 1 1 1 

60

115 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

179 1 1 1 1 2 1 1 2 1 1 1 1 1 

60

172 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

12

5

101 1 1 1 2 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

12

5

103 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 
B-1 

12

5

110 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

12

5

165 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 

12

5

107 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 

12

5

167 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

12

5

191 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 2 

12

5

106 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

12

5

160 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

12

5

163 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

12

5

112 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

12

5

166 1 1 1 1 1 1 1 1 1 

12

5

117 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

12

5

156 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 

12

5

158 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 

12

5

159 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

12

5

135 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

12

5

181 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

12

5

111 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

12

5

113 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

25

0

161 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 

25

0

152 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

25

0

107 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 

25

0

115 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 2 1 1 

25

0

114 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

25

0

159 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

25

0

193 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

25

0

171 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

25

0

154 1 1 1 1 1 1 1 1 1 1 1 1 1 

25

0

119 1 1 1 2 1 1 

25

0

156 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

25

0

134 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 
B-1 

25

0

110 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1

25

0

102 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

25

0

153 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

25

0

133 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

25

0

151 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 

25

0

113 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

25

0

122 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

25

0

136 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

F1 egg production

DOSE ID

#### 

#### 

#### 

10/

19 

10/

20 

10/

21 

10/

22 

week 16 

10/

23 

10/

24 

10/

25 

10/

26 

10/

27 

10/

28 

10/

29 

week 17 

10/

30 

10/

31 

11

/1 

11

/2 

11

/3 

11

/4 

11

/5 

week 18 

11

/6 

11

/7 

11

/8 

11

/9 

#### 

#### 

#### 

week 19 

#### 

0

217 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

211 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

234 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

237 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

235 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

245 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

219 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

242 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

226 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

209 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

215 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 

0

299 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

203 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

228 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

225 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

241 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

212 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

202 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

0

207 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 
B-1 

0

210

30

253 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

226 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

212 1 1 1 1 

30

242 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

202 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

230 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 

30

232 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

220 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

245 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

251 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

227 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 

30

250 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

247 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

211 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

231 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

235 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 

30

225 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30

206 1 1 1 1 1 1 1 1 1 1 

30

249 1 1 1 1 1 1 1 1 1 1 1 1 

30

221 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

162 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

166 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

182 1 1 1 1 2 1 

60

112 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

119 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

186 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

165 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

175 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

102 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 
B-1 

60

185 1 1 1 1 1 1 1 1 1 1 1 1 

60

184 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

181 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

183 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

174 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

189 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 

60

170 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

110 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

115 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

179 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

60

172 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 

12

5

101 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

12

5

103 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

12

5

110 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

12

5

165

12

5

107 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 

12

5

167 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

12

5

191 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

12

5

106 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

12

5

160 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

12

5

163 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

12

5

112 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

12

5

166 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

12

5

117 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

12

5

156 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

12

5

158 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

12

5

159 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

12

5

135 1 1 2 1 1 1 1 1 1 1 1 1 1 1 

12

5

181 2 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 2 1 1 1 1 1 

12

5

111 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 
B-1 

12

5

113 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 

25

0

161 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

25

0

152 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

25

0

107 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

25

0

115 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

25

0

114 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

25

0

159 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

25

0

193 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

25

0

171 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

25

0

154 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

25

0

119

25

0

156 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

25

0

134 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

25

0

110

25

0

102 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

25

0

153 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 

25

0

133 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

25

0

151 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 

25

0

113 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

25

0

122 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

25

0

136 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 
F1 egg production

DOSE ID #### #### #### 11/23 11/24 11/25 11/26 

week 

21 11/27 11/28 11/29 11/30 12/1 12/2 12/3 

week 

22 

0 217 1 1 1 2 1 

0 211 1 1 1 1 2 1 1 

0 234 1 1 1 

0 237 1 1 1 1 1 1 1 

0 235 1 1 1 1 

0 245 

0 219 1 1 1 1 2 

0 242 1 1 1 1 1 1 1 1 1 

0 226 1 1 1 1 1 1 1 1 1 1 2 

0 209 1 1 1 1 2 1 1 

0 215 1 1 1 1 1 1 1 2 1 

0 299 1 1 1 1 1 1 

0 203 1 1 1 1 1 1 

0 228 1 1 2 1 

0 225 1 1 2 1 1 

0 241 1 1 1 

0 212 1 1 1 1 1 1 

0 202 1 1 

0 207 1 1 1 1 1 1 1 

0 210 

30 253 1 1 1 1 1 1 1 1 

30 226 1 1 1 1 1 1 1 1 

30 212 1 1 1 1 1 1 1 

30 242 1 1 1 1 1 1 1 1 1 

30 202 1 1 1 1 1 1 1 1 1 1 1 1 

30 230 1 1 1 1 1 1 1 1 1 1 

30 232 1 1 1 1 1 1 2 1 1 

30 220 1 1 1 1 1 1 1 1 1 1 

30 245 1 1 2 1 1 1 1 1 1 

30 251 1 1 1 1 1 1 1 

30 227 1 1 1 1 1 1 1 1 

30 250 1 1 1 1 1 1 1 1 1 1 1 1 1 

30 247 1 1 1 1 1 1 1 1 1 

30 211 1 1 1 1 1 1 1 1 1 

30 231 1 1 1 1 1 1 1 1 1 1 

30 235 1 1 1 1 1 1 1 1 1 

30 225 1 1 1 1 1 1 2 1 1 1 

30 206 1 1 1 

30 249 1 1 1 1 1 1 1 1 1 1 

30 221 1 1 1 1 1 1 1 1 1 1 1 1 

60 162 1 1 1 1 1 

60 166 1 1 1 1 1 1 1 1 1 1 1 

60 182 

60 112 1 2 1 1 1 1 

60 119 

60 186 1 1 1 1 1 1 1 1 1 1 1 1 
B-1 

60 165 1 1 1 1 1 1 1 1 1 1 1 1 

60 175 1 1 1 1 1 1 1 1 

60 102 1 1 1 2 1 1 1 1 1 1 

60 185 1 2 1 1 1 

60 184 1 1 1 1 1 1 1 1 1 1 1 1 

60 181 1 

60 183 1 1 

60 174 1 2 1 

60 189 1 1 2 1 1 1 1 1 1 

60 170 1 1 1 1 1 1 1 1 1 1 1 

60 110 1 1 1 1 1 1 1 

60 115 1 1 1 

60 179 1 1 1 1 1 1 1 

60 172 2 1 1 1 1 

125 101 1 1 2 1 1 

125 103 1 1 1 1 1 1 2 1 

125 110 1 1 1 1 1 1 1 1 2 1 1 

125 165 1 

125 107 1 1 2 1 

125 167 1 1 1 1 1 1 1 1 1 1 1 1 

125 191 1 1 1 1 1 2 1 

125 106 1 1 1 1 1 1 1 1 1 1 1 1 

125 160 1 1 1 1 1 1 1 1 1 1 1 

125 163 1 1 1 1 1 1 1 2 1 1 1 1 

125 112 1 1 1 1 1 1 2 1 1 1 

125 166 1 1 1 2 1 1 1 1 1 1 

125 117 1 1 1 2 1 1 1 2 1 1 

125 156 1 1 1 1 1 1 1 1 1 1 1 

125 158 1 1 1 1 1 2 1 

125 159 1 1 1 1 1 1 1 1 1 1 1 

125 135 

125 181

125 111 1 1 1 1 1 1 1 

125 113 1 1 1 1 1 1 1 1 1 1 1 1 

250 161 1 1 1 1 1 1 1 1 1 1 

250 152 1 1 1 1 1 2 1 2 1 1 

250 107 1 1 1 1 

250 115 1 1 1 1 1 2 1 

250 114 1 1 1 1 1 1 1 1 1 1 1 

250 159 1 1 1 1 1 1 1 1 

250 193 1 1 1 1 1 1 1 1 1 

250 171 1 1 1 1 1 1 1 1 2 1 1 1 

250 154 1 1 1 

250 119 

250 156 1 1 1 1 1 1 1 

250 134 1 1 1 1 1 1 1 1 2 1 

250 110 

250 102 1 1 1 1 1 1 1 1 1 1 1 1 
B-1 

250 153 1 1 1 1 1 1 1 

250 133 1 1 1 1 1 1 1 1 1 1 

250 151 1 1 1 1 1 1 

250 113 1 1 1 1 1 1 1 1 

250 122 1 1 1 1 1 1 1 1 1 1 1 1 

250 136 1 1 1 1 1 1 1 1 1 1 1 1 

F2 cloaca measurements and foam 

 12/23 12/30 1/6 

DOSE ID CG area foam CG area foam CG area foam DOSE ID first day of foam 

0 193 25 N 25 Y 25 Y 0 193 1/19 

0 195 238 Y 210 Y 210 Y 0 195 12/23 

0 196 252 Y 255 Y 289 Y 0 196 12/23 

0 198 25 N 143 Y 169 Y 0 198 12/28 

0 184 270 Y 272 Y 240 Y 0 184 12/23 

0 192 120 Y 182 Y 210 Y 0 192 12/23 

0 177 25 N 25 N 169 N 0 177 

0 180 25 N 25 N 182 Y 0 180 1/5 

0 304 25 N 100 N 225 Y 0 304 12/31 

0 308 156 Y 240 Y 272 Y 0 308 12/23 

0 322 156 Y 196 Y 210 Y 0 322 12/23 

0 321 25 N 25 N 25 Y 0 321 1/6 

0 30 182 Y 450 Y 323 Y 0 301 12/23 

0 182 25 N 25 N 225 Y 0 182 1/3 

0 185 156 Y 156 Y 238 Y 0 185 12/23 

0 173 144 Y 156 Y 224 Y 0 173 12/23 

0 187 25 N 25 N 195 Y 0 187 1/3 

0 303 156 Y 182 Y 225 Y 0 303 12/23 

0 310 25 N 169 Y 225 Y 0 310 12/27 

0 329 25 N 169 Y 224 Y 0 329 12/29 

30 94 25 N 25 N 210 Y 30 94 1/6 

30 93 25 N 210 Y 255 Y 30 93 12/28 

30 90 25 N 25 N 210 Y 30 90 1/6 

30 80 156 Y 255 Y 272 Y 30 80 12/23 

30 78 196 Y 238 Y 255 Y 30 78 12/23 

30 76 25 N 182 Y 195 Y 30 76 12/28 

30 407 132 N 225 Y 255 Y 30 407 12/24 

30 402 256 Y 255 Y 255 Y 30 402 12/23 

30 417 25 N 25 N 196 Y 30 417 1/5 

30 406 169 Y 224 Y 255 Y 30 406 12/23 

30 92 25 N 240 N 304 N 30 92 12/28 

30 88 25 N 25 Y 25 Y 30 88 

30 95 25 N 25 N 25 N 30 95 

30 91 210 Y 270 Y 288 Y 30 91 12/23 

30 77 156 Y 182 Y 240 Y 30 77 12/23 

30 83 182 Y 210 Y 288 Y 30 83 12/23 

30 404 25 N 169 Y 270 Y 30 404 

30 100 132 Y 196 Y 240 Y 30 100 12/23 
B-1 

30 99 169 Y 210 Y 240 Y 30 99 12/23 

30 97 25 N 25 N 88 Y 30 97 1/4 

60 90 182 Y 195 Y 210 Y 60 90 12/23 

60 88 25 N 210 Y 210 Y 60 88 

60 205 25 Y 195 Y 255 Y 60 205 12/23 

60 208 169 Y 255 Y 285 Y 60 208 12/23 

60 81 144 Y 195 Y 210 Y 60 81 12/23 

60 84 168 Y 210 Y 238 Y 60 84 12/23 

60 80 182 Y 210 Y 240 Y 60 80 12/23 

60 86 144 Y 195 Y 182 Y 60 86 12/23 

60 96 25 N 25 Y 195 Y 60 96 1/3 

60 79 25 N 182 Y 196 Y 60 79 12/28 

60 203 25 N 182 Y 210 Y 60 203 12/28 

60 95 25 N 25 N 210 Y 60 95 1/5 

60 99 25 N 25 Y 182 Y 60 99 1/3 

60 98 25 N 25 N 182 Y 60 98 1/4 

60 201 25 N 25 N 30 N 60 201 

60 204 25 N 196 Y 240 Y 60 204 12/28 

60 209 156 Y 196 Y 225 Y 60 209 12/23 

60 210 25 N 132 N 156 N 60 210 12/28 

60 219 25 N 182 Y 272 Y 60 219 12/27 

60 218 90 N 196 Y 272 Y 60 218 12/24 

125 75 169 Y 210 Y 240 Y 125 75 12/23 

125 77 168 Y 195 Y 240 Y 125 77 12/23 

125 95 144 Y 210 Y 255 Y 125 95 12/23 

125 233 25 N 25 N 182 Y 125 233 1/5 

125 100 144 Y 225 Y 288 Y 125 100 12/23 

125 81 25 N 169 Y 225 Y 125 81 12/27 

125 98 121 Y 169 Y 210 Y 125 98 12/23 

125 97 25 N 182 Y 225 Y 125 97 12/28 

125 208 25 N 169 Y 224 Y 125 208 12/27 

125 76 156 Y 210 Y 238 Y 125 76 12/23 

125 235 25 N 210 Y 288 Y 125 235 12/27 

125 93 182 Y 169 Y 210 Y 125 93 12/23 

125 82 156 Y 196 Y 272 Y 125 82 12/23 

125 94 182 Y 169 Y 240 Y 125 94 12/23 

125 90 156 Y 182 Y 224 Y 125 90 12/23 

125 99 25 N 25 N 210 Y 125 99 1/3 

125 78 25 N 25 N 240 Y 125 78 1/5 

125 85 144 Y 182 Y 270 Y 125 85 12/23 

125 204 25 N 25 N 210 Y 125 204 1/4 

125 79 25 N 25 N 25 N 125 79 

250 84 182 Y 195 Y 270 Y 250 84 12/23 

250 83 156 Y 196 Y 240 Y 250 83 12/23 

250 87 25 N 182 Y 224 Y 250 87 12/28 

250 91 100 Y 196 Y 210 Y 250 91 12/23 

250 77 25 N 210 Y 270 Y 250 77 12/28 

250 76 25 N 182 Y 225 Y 250 76 12/28 
B-1 

250 98 156 Y 182 Y 210 Y 250 98 12/23 

250 100 25 N 25 N 25 N 250 100 

250 95 25 N 169 Y 195 Y 250 95 12/27 

250 99 25 N 25 N 25 N 250 99 

250 231 25 N 25 N 25 N 250 231 

250 189 144 Y 210 Y 238 Y 250 189 12/23 

250 230 25 N 25 Y 210 Y 250 230 1/4 

250 90 25 N 182 Y 272 Y 250 90 12/28 

250 75 25 N 210 Y 255 Y 250 75 12/28 

250 81 143 Y 182 Y 240 Y 250 81 12/23 

250 85 25 N 25 Y 210 Y 250 85 1/3 

250 80 144 Y 132 Y 225 Y 250 80 12/23 

250 181 169 Y 255 Y 255 Y 250 181 12/23 

250 190 25 N 169 Y 210 Y 250 190 12/28 

F2 egg strength

DATE DOSE ID Thickness 1 Thickness 2 Strength Yolk DATE Thickness 1 Thickness 2 Strength DATE Thickness 1 Thickness 2 Strength

 0 183 

 0 199 

1/26 0 200 0.43 0.6 1.414 1/28 0.3 0.37 1.364 

1/27 0 194 0.36 0.41 1.193 

1/25 0 179 0.5 0.34 0.729 1/27 1.04 0.86 0.495 1/28 0.58 0.42 0.93 

1/25 0 191 0.4 0.35 0.761 1/27 0.46 0.3 0.691 1/28 0.33 0.35 0.345 

1/28 0 188 0.37 0.34 0.463 1/26 0.37 0.4 1.232 

1/25 0 189 0.39 0.43 0.939 

1/27 0 313 0.35 0.41 0.598 

1/25 0 174 0.54 0.49 0.696 N 1/26 0.45 0.37 0.382 1/27 0.48 0.5 1.071 

1/25 0 314 0.45 0.49 1.124 1/26 0.43 0.38 1.02 

1/25 0 309 0.31 0.36 0.785 1/26 0.68 0.7 0.757 

1/25 0 186 0.55 0.38 0.931 1/26 0.42 0.33 1.371 

1/25 0 197 0.49 0.62 1.155 1/26 0.54 0.54 1.439 

1/25 0 178 0.62 0.62 1.017 1/26 0.76 0.84 0.409 

 0 181 

1/26 0 175 0.78 0.44 0.851 1/27 0.44 0.45 0.699 1/28 0.59 0.61 0.764 

 0 190 

1/25 0 323 0.43 0.37 0.953 1/26 0.52 0.48 1.551 

1/25 0 305 0.39 0.47 0.7 

1/25 30 81 0.37 0.38 1.055 1/26 0.57 0.54 1.004 

1/25 30 96 0.51 0.36 1.172 1/27 0.5 0.42 1.981 1/28 0.38 0.34 1.503 

1/26 30 89 0.35 0.38 1.534 1/27 0.65 0.47 1.003 

1/25 30 85 0.41 0.43 0.911 1/26 0.51 0.38 0.778 

1/27 30 403 0.66 0.78 0.738 

1/25 30 405 1.06 0.71 1.249 1/26 0.57 0.64 1.252 

1/25 30 408 0.62 0.45 1.215 1/26 0.53 0.41 1.157 

1/26 30 427 0.38 0.41 1.118 1/27 0.37 0.44 1.185 1/28 0.42 0.39 0.393 

1/26 30 409 0.41 0.47 1.002 1/28 0.35 0.32 0.72 

 30 432 

 30 84 1/26 0.72 0.53 1.105 
B-1 

1/25 30 86 0.35 0.3 1.257 1/28 0.35 0.28 1.024 

1/25 30 82 0.49 0.45 1.02 1/26 0.42 0.68 0.996 

1/25 30 87 0.43 0.45 1.027 1/28 0.39 0.33 1.106 

1/25 30 401 0.44 0.56 0.943 N 1/26 0.3 0.25 1.024 1/27 0.89 0.9 0.977 

1/25 30 74 0.6 0.48 1.528 1/26 0.72 0.52 

1/25 30 73 0.65 0.41 0.792 1/26 0.34 0.45 

1/27 30 86 0.52 0.45 1.065 

1/25 30 75 0.41 0.42 1.096 1/26 0.43 0.4 1.074 

 30 79 

1/25 60 75 0.91 0.84 0.987 1/26 0.57 0.75 0.764 

1/25 60 91 0.45 0.46 1.344 1/26 0.33 0.35 1.11 

1/25 60 89 0.43 0.47 1.707 1/27 0.4 0.35 0.34 1/28 0.32 0.34 1 

1/25 60 73 0.41 0.38 1.47 N 1/28 0.45 0.39 1.447 

1/28 60 85 0.39 0.34 1.149 

1/25 60 100 0.29 0.28 1.214 1/26 0.39 0.41 1.299 

1/25 60 94 0.42 0.41 0.834 1/26 0.35 0.42 1.175 

1/25 60 76 0.3 0.41 0.35 1/26 0.87 0.81 1.452 

 60 78 

1/25 60 93 0.6 0.45 1.358 1/26 0.81 1.05 1.091 

1/25 60 82 0.56 0.35 0.974 1/26 0.83 0.97 0.911 

1/25 60 74 0.65 0.63 1.781 1/26 0.41 0.42 1.228 

1/25 60 92 0.53 0.56 0.292 1/26 0.87 0.57 0.871 

 60 87 

1/25 60 202 0.44 0.56 0.887 1/27 0.42 0.36 1.157 1/28 0.38 0.31 0.822 

1/25 60 77 0.71 0.89 1.3 1/26 0.42 0.38 1.231 

1/25 60 222 0.32 0.36 1.562 1/26 0.61 0.85 1.493 

1/25 60 97 0.74 0.5 1.158 

1/26 60 215 0.6 0.48 0.333 1/27 0.3 0.32 0.777 1/28 0.32 0.35 0.425 

1/25 60 216 0.37 0.26 0.347 

1/25 125 74 0.52 0.41 0.935 1/27 0.47 0.36 0.898 1/28 0.39 0.35 1.12 

1/25 125 80 0.42 0.34 0.979 1/26 0.62 0.65 1.369 

1/26 125 83 0.67 0.6 0.902 1/27 0.62 0.6 1.041 

1/25 125 73 0.36 0.32 0.45 1/28 0.38 0.33 0.967 

1/25 125 206 0.61 0.4 0.924 1/27 0.29 0.32 0.808 

1/25 125 205 0.37 0.33 0.921 1/26 0.64 0.73 0.789 

1/25 125 234 0.32 0.33 1.152 1/26 0.46 0.53 1.353 

1/25 125 201 0.42 0.37 0.495 1/27 0.45 0.47 0.557 1/28 0.28 0.32 1.392 

1/27 125 239 0.52 0.46 0.559 

1/25 125 237 0.45 0.59 0.821 1/26 0.36 0.41 1.557 

1/26 125 88 0.72 0.78 0.476 1/28 0.27 0.37 0.464 1/28 0.4 0.42 0.75 

 125 87 1/26 0.39 0.42 1.602 

1/25 125 89 0.4 0.39 1.181 1/26 0.47 0.67 1.315 

1/25 125 84 0.53 0.43 1.287 1/26 0.6 0.48 1.227 

1/25 125 207 0.64 0.48 1.31 1/26 0.69 0.57 1.296 

1/25 125 202 0.4 0.45 1.325 1/26 0.61 0.67 1.107 

1/25 125 259 0.52 0.39 1.004 1/26 0.3 0.32 0.829 

1/25 125 209 0.88 0.65 1.042 1/26 0.52 0.36 1.318 

1/25 125 258 0.48 0.35 1.153 1/26 0.62 0.61 0.819 
B-1 

1/25 125 260 0.34 0.39 1.013 1/26 0.37 0.42 1.101 

1/25 250 89 0.49 0.33 1.182 1/26 0.65 0.62 1.322 

1/25 250 82 0.77 0.8 0.909 1/26 0.35 0.46 1.414 

1/25 250 79 0.37 0.38 1.233 1/26 0.41 0.43 1.555 

 250 92 

1/25 250 94 0.41 0.45 1.33 1/26 0.61 0.53 1.582 

1/25 250 228 0.46 0.53 1.258 1/26 0.47 0.37 0.835 

 250 188 

1/25 250 184 0.4 0.59 1.342 1/26 0.43 0.39 1.081 

1/25 250 97 0.62 0.72 0.332 1/26 0.53 0.64 1.181 

1/25 250 74 0.48 0.52 1.623 1/26 0.32 0.39 1.316 

1/25 250 78 0.63 0.72 1.046 1/26 0.47 0.42 1.229 

1/25 250 88 0.48 0.44 0.996 1/26 0.38 0.4 0.89 

1/25 250 96 0.59 0.42 0.357 1/26 0.34 0.37 1.397 

1/25 250 93 0.29 0.38 1.119 1/26 0.48 0.43 1.305 

1/26 250 225 0.41 0.55 0.86 1/27 0.56 0.58 0.683 

1/25 250 221 0.46 0.49 1.358 1/26 0.38 0.46 1.294 

F2 sperm measurements 

ID Dose Volume Weight # sperm/gram % Motile % Progressive 

182 0 0.5 0.019 178.9 42 4 

182 0 0.5 0.019 151.3 27 1 

303 0 0.5 0.019 31 61 6 

303 0 0.5 0.019 36.1 48 10 

303 0 0.5 0.019 27.5 44 0 

173 0 0.5 0.02 313.7 42 8 

173 0 0.5 0.02 398.6 66 11 

173 0 0.5 0.02 346.4 47 11 

329 0 0.5 0.015 8200.5 98 37 

329 0 0.5 0.015 7125.3 97 35 

329 0 0.5 0.015 4501.2 94 26 

198 0 0.5 0.016 5125.3 90 15 

198 0 0.5 0.016 7209.6 92 21 

198 0 0.5 0.016 8593.4 93 19 

195 0 0.5 0.011 9244.2 94 23 

195 0 0.5 0.011 6063.4 94 20 

195 0 0.5 0.011 5144 96 19 

192 0 0.5 0.011 12151.7 97 20 

192 0 0.5 0.011 8573.3 94 21 

192 0 0.5 0.011 7802.9 95 14 

308 0 0.5 0.008 9328.1 92 20 

308 0 0.5 0.008 9157.2 88 18 

308 0 0.5 0.008 8371.3 84 17 

321 0 0.5 0.006 6195.9 85 13 

321 0 0.5 0.006 6514.8 82 15 

321 0 0.5 0.006 4647 55 7 
B-1 

180 0 0.5 0.008 8200.5 83 21 

180 0 0.5 0.008 9430.6 91 20 

180 0 0.5 0.008 8918 86 14 

93 30 0.5 0.018 319.5 30 0 

93 30 0.5 0.018 283.2 38 5 

76 30 0.5 0.02 35.9 73 0 

76 30 0.5 0.02 29.4 72 6 

76 30 0.5 0.02 39.2 88 4 

80 30 0.5 0.019 39.6 74 9 

80 30 0.5 0.019 48.2 89 21 

80 30 0.5 0.019 34.4 60 5 

402 30 0.5 0.008 3006.9 67 8 

402 30 0.5 0.08 4100.3 84 11 

402 30 0.5 0.008 3656.1 82 7 

88 30 0.5 0.008 7927.2 95 14 

88 30 0.5 0.008 7141.3 94 12 

88 30 0.5 0.008 9191.4 96 9 

91 30 0.5 0.008 10489.8 89 12 

91 30 0.5 0.008 7483 85 13 

91 30 0.5 0.008 9054.7 91 12 

406 30 0.5 0.009 1974.2 58 5 

406 30 0.5 0.009 4221.7 59 9 

406 30 0.5 0.009 5710 74 13 

97 30 0.5 0.009 1549 57 10 

97 30 0.5 0.009 1640.1 74 20 

97 30 0.5 0.009 2460.2 81 10 

83 30 0.5 0.008 9840.6 93 29 

83 30 0.5 0.008 12027.4 95 32 

83 30 0.5 0.008 9943.1 92 27 

100 30 0.5 0.008 10353.1 95 19 

100 30 0.5 0.008 13940.9 94 18 

100 30 0.5 0.008 13325.8 95 18 

79 60 0.5 0.014 420.1 64 9 

79 60 0.5 0.014 373.4 60 20 

79 60 0.5 0.014 560.2 75 10 

79 60 0.5 0.014 2050.1 86 25 

79 60 0.5 0.014 8630.1 89 20 

84 60 0.5 0.014 9489.2 91 15 

84 60 0.5 0.014 6658 81 10 

84 60 0.5 0.014 6130.9 86 13 

86 60 0.4 0.013 11459.7 92 21 

86 60 0.5 0.013 10429.4 93 17 

86 60 0.5 0.013 10744.8 91 20 

95 60 0.5 0.006 9020.6 90 12 

95 60 0.5 0.006 9248.3 93 23 

95 60 0.5 0.006 5831.5 77 16 

98 60 0.5 0.008 7483 85 19 

98 60 0.5 0.008 6047.9 91 18 
B-1 

98 60 0.5 0.008 5740.4 93 15 

204 60 0.5 0.009 7167.8 90 16 

204 60 0.5 0.009 5254.4 82 10 

204 60 0.5 0.009 6712.3 85 14 

88 60 0.5 0.007 12457 88 19 

88 60 0.5 0.007 13042.7 88 20 

88 60 0.5 0.007 9567.3 90 13 

218 60 0.5 0.008 4271.1 82 12 

218 60 0.5 0.008 3861.1 90 17 

218 60 0.5 0.008 3826.9 76 9 

208 60 0.5 0.007 7458.6 93 19 

208 60 0.5 0.007 8512.9 93 22 

208 60 0.5 0.007 6091.8 90 10 

210 60 0.5 0.007 8708.2 84 10 

210 60 0.5 0.007 5701.3 81 10 

210 60 0.5 0.007 7107.1 85 14 

81 125 0.5 0.02 3362.2 91 18 

81 125 0.5 0.02 3854.2 90 15 

81 125 0.5 0.02 4018.2 90 22 

77 125 0.5 0.015 4391.8 87 14 

77 125 0.5 0.015 4045.6 79 11 

77 125 0.5 0.015 5703.9 79 9 

233 125 0.5 0.019 4675.7 92 15 

233 125 0.5 0.019 4013.9 90 10 

233 125 0.5 0.019 3913.2 86 15 

76 125 0.5 0.005 5303 79 10 

76 125 0.5 0.005 5849.7 87 9 

76 125 0.5 0.005 5084.3 80 9 

97 125 0.5 0.008 6492.1 89 17 

97 125 0.5 0.008 6389.6 84 7 

97 125 0.5 0.008 7312.1 80 14 

93 125 0.5 0.008 8610.5 92 14 

93 125 0.5 0.008 6526.2 86 11 

93 125 0.5 0.008 6082 89 12 

94 125 0.5 0.005 4100.3 80 15 

94 125 0.5 0.005 3006.9 67 13 

94 125 0.5 0.005 4647 66 7 

79 125 0.5 0.007 3241.2 55 5 

79 125 0.5 0.007 6794.7 86 9 

79 125 0.5 0.007 4295.5 62 5 

85 125 0.5 0.007 5467 62 11 

85 125 0.5 0.007 5232.7 81 14 

85 125 0.5 0.007 8786.3 80 8 

99 125 0.5 0.006 11754.1 90 19 

99 125 0.5 0.006 9795 89 19 

99 125 0.4 0.006 7157.7 75 8 

81 250 0.5 0.013 9146.7 93 20 

81 250 0.5 0.013 10744.8 95 18 
B-1 

81 250 0.5 0.013 10892 94 16 

90 250 0.5 0.009 880.8 17 0 

90 250 0.5 0.009 850.4 54 4 

90 250 0.5 0.009 789.7 58 0 

80 250 0.5 0.013 10303.2 90 17 

80 250 0.5 0.013 10807.8 93 14 

80 250 0.5 0.013 10050.9 93 13 

91 250 0.5 0.01 6587.7 89 17 

91 250 0.5 0.01 7517.1 89 18 

91 250 0.5 0.01 6806.4 94 12 

190 250 0.5 0.011 2982 61 7 

190 250 0.5 0.011 4249.4 85 12 

190 250 0.5 0.011 4547.6 79 15 

76 250 0.5 0.011 7305.9 93 31 

76 250 0.5 0.011 8051.4 96 33 

76 250 0.5 0.011 8076.3 90 26 

83 250 0.5 0.008 4066.1 73 12 

83 250 0.5 0.008 5262 87 13 

83 250 0.5 0.008 4339.4 84 7 

100 250 0.5 0.007 7653.8 81 18 

100 250 0.5 0.007 6248 68 13 

100 250 0.5 0.007 4490.8 59 11 

189 250 0.5 0.01 1503.4 11 0 

189 250 0.5 0.01 1312.1 8 0 

189 250 0.5 0.01 1530.8 5 0 

99 250 0.5 0.005 273.4 0 0 

99 250 0.5 0.005 710.7 46 0 

99 250 0.5 0.005 546.7 0 0 

F2 growth measurements 

11/2 

DOSE

baby

id BW

tarsus

1

tarsus

2

avg

tarsus

culmen

1

culmen

2  

avg

culmen

0 394 8 14.9 14.4 14.65 4.9 5.3 5.1 

0 392 9 17.8 17.4 17.6 4 4.1 4.05 

0 401 7 15.9 16.3 16.1 4.1 4.5 4.3 

0 391 8 16.4 16.5 16.45 4.2 4.5 4.35 

0 396 19 16.1 16.2 16.15 4.2 4.6 4.4 

0 388 9 14.6 14.3 14.45 4.2 4.4 4.3 

0 403 6 13.5 13.8 13.65 3 3 3 

0 395 7 17.2 17.5 17.35 3.4 3.1 3.25 

0 398 8 15.4 15.5 15.45 3.7 3.9 3.8 

0 404 8 16.1 16.6 16.35 2.7 3 2.85 

0 426 7 13.4 13.9 13.65 4.5 4.1 4.3 

0 432 6 13.2 13.4 13.3 3.2 3.6 3.4 

0 424 6 16.2 16.2 16.2 3 3.2 3.1 

0 431 6 15.8 16.1 15.95 3.5 3.6 3.55 

0 440 6 16 16.1 16.05 3.5 3.6 3.55 
B-1 

0 421 7 16 16.3 16.15 4 4.1 4.05 

0 439 6 16.1 16.5 16.3 3.8 4 3.9 

0 437 6 15.7 15.6 15.65 3.8 3.9 3.85 

0 436 6 15.2 15.3 15.25 3.5 3.8 3.65 

0 420 7 15.5 15.7 15.6 3 3.2 3.1 

30 446 7 13.5 12.6 13.05 3.1 2.8 2.95 

30 450 7 12.9 13 12.95 4.2 3.8 4 

30 456 8 14.5 15 14.75 3.9 3.9 3.9 

30 454 8 16.3 15.6 15.95 4.1 3.9 4 

30 455 9 17 17.3 17.15 3.7 3.7 3.7 

30 451 6 15.6 15.9 15.75 3.8 4 3.9 

30 457 7 12.3 12.5 12.4 3.2 3.7 3.45 

30 460 9 18 18.3 18.15 3.1 3.4 3.25 

30 445 7 15.5 15.9 15.7 3.9 4.2 4.05 

30 453 9 18.2 18.4 18.3 3.3 3.6 3.45 

30 393 7 16.6 17 16.8 3.3 3.5 3.4 

30 487 8 17.1 17.4 17.25 3.8 4.1 3.95 

30 486 6 16.2 15.9 16.05 4 4.2 4.1 

30 492 6 14.7 14.9 14.8 3.8 4.1 3.95 

30 484 6 16.4 16.6 16.5 3.8 3.9 3.85 

30 489 7 17.1 17.2 17.15 3.6 3.7 3.65 

30 493 7 14 14.2 14.1 3.5 3.8 3.65 

30 491 7 17.4 17.1 17.25 3.8 4.1 3.95 

30 495 7 16.8 17.1 16.95 3.8 4 3.9 

30 490 6 14.8 15 14.9 3.4 3.6 3.5 

60 373 8 15.3 15.6 15.45 4 3.6 3.8 

60 375 9 17 17.6 17.3 4.6 4.3 4.45 

60 363 10 16.5 16.3 16.4 3.8 3.5 3.65 

60 359 9 16.7 16.9 16.8 4.3 4.4 4.35 

60 365 10 15.9 15.7 15.8 4.2 4.1 4.15 

60 370 10 17.4 17.6 17.5 3.7 4.1 3.9 

60 372 9 13 13.1 13.05 4.2 4.2 4.2 

60 376 6 15.8 16 15.9 3 3.4 3.2 

60 377 6 15.7 15.9 15.8 3.7 3.8 3.75 

60 364 7 17.4 17.5 17.45 3.7 3.8 3.75 

60 399 7 16 16.4 16.2 3.6 3.9 3.75 

60 321 6 16.4 16.6 16.5 4.2 4.4 4.3 

60 400 8 16.5 15.7 16.1 3.8 3.9 3.85 

60 397 8 16.1 16.3 16.2 3.4 3.6 3.5 

60 396 6 16.7 16.8 16.75 3.3 3.4 3.35 

60 395 7 15.5 15.7 15.6 4.1 4.3 4.2 

60 394 6 15.7 15.8 15.75 3.6 3.6 3.6 

60 398 7 14.9 15.2 15.05 3.7 3.9 3.8 

125 387 10 17.6 17 17.3 4.9 4.4 4.65 

125 382 7 15.1 15.7 15.4 3.2 3.7 3.45 

125 391 9 13.2 13.9 13.55 3.3 3.7 3.5 

125 392 9 15.7 15.2 15.45 3.5 3.7 3.6 

125 386 9 16.6 17 16.8 3.8 4 3.9 
B-1 

125 389 7 16.4 16.5 16.45 3.4 3.8 3.6 

125 396 8 14.9 15.2 15.05 3.2 3.8 3.5 

125 398 7 15.7 16.1 15.9 3.8 3.8 3.8 

125 395 8 15.3 15.6 15.45 4.2 4 4.1 

125 388 10 15.4 15.1 15.25 4.1 3.8 3.95 

125 356 7 18.3 18 18.15 3.6 3.8 3.7 

125 354 7 15.8 16 15.9 3.5 3.7 3.6 

125 353 8 14.7 15 14.85 3.6 3.7 3.65 

125 356 8 16.4 16.5 16.45 4.1 4.5 4.3 

125 357 6 15.8 16 15.9 3.7 3.4 3.55 

125 359 6 15.7 15.9 15.8 3.1 3.3 3.2 

125 361 6 15.3 15.4 15.35 3.9 3.4 3.65 

125 362 7 16 16.2 16.1 3.4 3.6 3.5 

125 363 7 14.8 15 14.9 3.8 3.4 3.6 

125 352 7 16 15.8 15.9 4 4.2 4.1 

250 415 11 16.3 16.6 16.45 4.7 4.9 4.8 

250 420 10 16.8 17.2 17 4.5 4.8 4.65 

250 422 7 16.7 16.9 16.8 3.8 4.1 3.95 

250 412 11 17 16.8 16.9 4 4.3 4.15 

250 427 7 13.3 13.6 13.45 3.6 3.8 3.7 

250 419 8 15.2 15.4 15.3 3.8 4.1 3.95 

250 425 6 13.4 13.7 13.55 3.5 3.6 3.55 

250 414 10 15.5 15.8 15.65 3.2 3.6 3.4 

250 418 8 18 18.4 18.2 4.1 4.2 4.15 

250 416 7 17.5 17.7 17.6 4 4.2 4.1 

250 445 8 15.8 16.1 15.95 4.1 4.3 4.2 

250 453 7 14.5 14.7 14.6 3.6 3.8 3.7 

250 455 6 15.8 16.2 16 3.8 3.9 3.85 

250 448 7 16.3 16.4 16.35 3.6 3.9 3.75 

250 450 7 14 14.1 14.05 4.1 4.4 4.25 

250 444 6 17.1 17.3 17.2 4.1 4.5 4.3 

250 460 6 16.3 16.4 16.35 3.6 3.8 3.7 

250 458 7 16.4 16.5 16.45 4 4.1 4.05 

250 452 6 15.6 15.9 15.75 3.6 3.7 3.65 

250 469 6 16 16.4 16.2 3.2 3.4 3.3 
F2 growth measurements 

11/6 11/19 

DOSE

babyid BW

tarsus1

tarsus

2

avgtarsus

culmen1

culmen

2

avgculmen DOSE

babyid BW

tarsus1

tarsus2

avgtarsus

culmen1

culmen2

avgculmen

0 388 19 6.2 6.4 6.3 19.9 20 19.95 0 397 30 5.7 5.9 5.8 30.9 21 25.95 0 395 19 3.3 3.5 3.4 190.6 19.8 105.2 0 394 22 5.1 5 5.05 25.5 25.2 25.35 0 389 16 5 5.3 5.15 18.8 19.2 19 0 401 23 5.8 5.9 5.85 22.7 23 22.85 0 396 18 5.1 5.2 5.15 19.3 19.4 19.35 0 396 25 6.3 6.6 6.45 19.2 19.4 19.3 0 402 13 4.5 4.7 4.6 18.1 18.5 18.3 0 403 25 5.1 4.8 4.95 19.4 19.5 19.45 0 404 19 4.8 4.9 4.85 17.7 17.9 17.8 0 403 22 5.2 5.6 5.4 16.8 17 16.9 0 394 17 5.6 5.6 5.6 19.9 20.1 20 0 405 22 6 6.1 6.05 18.4 18.6 18.5 0 427 8 3.1 3.4 3.25 12.3 12.4 12.35 0 404 27 5.6 5.7 5.65 20.8 20.9 20.85 0 401 15 4.3 4.1 4.2 18.4 18.6 18.5 0 393 26 5.4 5.5 5.45 19.9 20.3 20.1 0 392 18 4 4.1 4.05 17.8 18 17.9 0 392 26 4.9 5.2 5.05 19.4 19.7 19.55 

30 496 15 4.7 4.5 4.6 21.8 21.6 21.7 30 455 82 5.4 5.7 5.55 25.5 25.3 25.4 30 453 19 4.4 4.5 4.45 21.2 20.9 21.05 30 444 29 5.7 5.8 5.75 24.7 25 24.85 30 456 12 4.3 4.3 4.3 17.4 17.6 17.5 30 447 27 5.8 6.1 5.95 25.1 25.3 25.2 30 457 14 5.1 5.2 5.15 18.4 18.7 18.55 30 453 26 5.2 5.3 5.25 23.8 23.9 23.85 30 444 20 4.8 5 4.9 20.1 20.2 20.15 30 449 27 5.8 5.8 5.8 22 22.2 22.1 30 448 21 5.2 5.3 5.25 21.1 21.2 21.15 30 454 25 6.1 6.2 6.15 17.9 18.2 18.05 30 455 20 4.4 4.7 4.55 20.5 20.7 20.6 30 458 28 5.3 5.4 5.35 22.1 22.2 22.15 30 451 14 4.4 4.3 4.35 16.4 16.8 16.6 30 446 23 4.7 4.8 4.75 17.1 17.3 17.2 30 445 15 4.7 4.9 4.8 18.1 18.3 18.2 30 457 23 5.5 5.6 5.55 17.8 17.9 17.85 30 458 19 4.1 4.2 4.15 18.4 18.5 18.45 30 451 21 5.7 6 5.85 16.8 17 16.9 60 369 19 5.2 5.4 5.3 20 20.2 20.1 60 370 25 6.1 6.3 6.2 22.2 22.3 22.25 60 370 17 4.8 4.9 4.85 19.6 19.9 19.75 60 365 29 6.1 6.5 6.3 26.3 26.7 26.5 60 376 18 4.6 4.7 4.65 19 19.5 19.25 60 366 28 4.2 4.5 4.35 22.9 23.4 23.15 60 378 14 4.7 4.5 4.6 18 18.5 18.25 60 367 23 5.6 5.6 5.6 20.8 20.9 20.85 60 367 17 5.3 5.5 5.4 18.5 18.4 18.45 60 362 29 5.2 5.5 5.35 23.5 23.7 23.6 60 381 15 4.6 4.6 4.6 20 20.1 20.05 60 375 29 5.3 5.6 5.45 20.5 20.7 20.6 60 366 19 5 5.3 5.15 19.6 19.7 19.65 60 377 24 5.1 5.3 5.2 17.6 17.9 17.75 60 362 19 5.6 5.7 5.65 20 19.7 19.85 60 361 24 5.8 5.9 5.85 20.2 20 20.1 60 373 16 4.1 4.4 4.25 20 20 20 60 379 30 6 6.2 6.1 19.1 18.8 18.95 60 369 18 5 4.8 4.9 15.7 16 15.85 60 380 25 6.7 6.8 6.75 19.4 19.6 19.5 

125 381 20 4.8 5 4.9 21.8 23 22.4 125 395 23 6.2 6.4 6.3 23.2 23.4 23.3 
B-1 

125 380 19 5.1 5.2 5.15 22.2 22.5 22.35 125 384 27 6.6 6.4 6.5 21.1 21.3 21.2 125 396 19 4.7 4.7 4.7 21.4 21.7 21.55 125 396 27 5.8 5.9 5.85 19.6 20 19.8 125 398 17 4.4 4.6 4.5 19.7 19.9 19.8 125 366 23 6.4 6.8 6.6 18.2 18.4 18.3 125 382 19 3.8 4 3.9 17.5 17.7 17.6 125 383 26 5.1 5.2 5.15 18.9 19.2 19.05 125 375 16 6.1 6.3 6.2 21.1 21.2 21.15 125 381 26 6.4 6.6 6.5 18.1 18.4 18.25 125 394 13 4.6 4.8 4.7 18.6 18.7 18.65 125 399 23 5.5 5.4 5.45 17.8 17.7 17.75 125 399 16 4.8 5 4.9 19.7 19.9 19.8 125 390 24 5.3 5.4 5.35 18 18.1 18.05 125 345 15 5.2 5.1 5.15 17.2 17.3 17.25 125 397 24 5.3 5.6 5.45 16.9 17.2 17.05 125 400 17 4.7 4.8 4.75 17.7 17.9 17.8 125 345 23 6.1 6.2 6.15 17.6 17.7 17.65 250 414 4.7 4.8 4.8 20.1 20.2 20.15 250 418 24 5.4 5.5 5.45 22.8 23.1 22.95 250 420 21 4.7 5 4.85 21.1 21.5 21.3 250 422 26 5.2 5.3 5.25 22.2 22.5 22.35 250 426 17 5.2 5 5.1 19 19.3 19.15 250 415 30 5.7 5.4 5.55 22 22.4 22.2 250 413 21 4.7 4.6 4.65 22.2 22.4 22.3 250 412 30 4.4 4.9 4.65 22.4 21.9 22.15 250 417 13 4.8 5 4.9 18.5 18.7 18.6 250 427 23 5.2 5.3 5.25 18.2 18.4 18.3 250 418 15 4.8 5 4.9 16.1 16.4 16.25 250 425 19 5.3 5.6 5.45 17.5 17.8 17.65 250 427 15 4.7 4.9 4.8 15.9 16 15.95 250 426 24 6.3 6.2 6.25 18.3 18.4 18.35 250 412 22 4.8 5 4.9 22 22.2 22.1 250 413 31 6.4 6.5 6.45 19.8 20.1 19.95 250 422 17 4.4 4.7 4.55 16.5 16.9 16.7 250 428 11 4.8 5.1 4.95 16.8 17 16.9 250 429 17 4.8 5 4.9 16.6 16.7 16.65 250 413 21 5.6 5.9 5.75 20 20.3 20.15 

F2 growth measurements 

11/23 11/27 11/30 

DOSE baby id ID BW tarsus 1 tarsus 2 avg tarsus culmen 1 culmen 2 avg culmen DOSE ID BW tarsus 1 tarsus 2 avg tarsus culmen 1 culmen 2 avg culmen DOSE ID BW tarsus 1 tarsus 2 avg tarsus 

0 393 191 38.3 6.8 6.7 6.75 22.5 22.9 22.7 0 192 52 7.1 7.3 7.2 29.2 30.3 29.75 0 183 56 7.8 8 7.9 0 392 192 38 4.8 5.1 4.95 24.5 24.7 24.6 0 195 48 7 7.3 7.15 27.5 27.6 27.55 0 200 57 7.9 8 7.95 0 402 193 30.3 6.9 7.2 7.05 23 22.7 22.85 0 191 49 6.5 6.9 6.7 27.4 27.5 27.45 0 181 65 7.2 7.5 7.35 0 394 194 35.8 6.1 6 6.05 23.3 23.7 23.5 0 185 49 7.3 7.4 7.35 28.9 29 28.95 0 197 58 6.9 7.1 7 0 356 195 36.2 6.6 6.8 6.7 21.7 21.8 21.75 0 193 42 6.1 6.5 6.3 27.8 27.7 27.75 0 196 52 6.8 6.9 6.85 0 397 196 29.3 7.6 7.8 7.7 21.2 21.3 21.25 0 196 40 6.1 6.5 6.3 24.6 24.9 24.75 0 198 57 8 8.1 8.05 0 1 197 33.6 6.1 6.4 6.25 25.5 24.9 25.2 0 194 49 7.1 7.3 7.2 27.1 27.3 27.2 0 182 47 6.2 6.3 6.25 0 389 198 34.3 5.4 5.3 5.35 22.4 22.7 22.55 0 184 48 5.9 6 5.95 27.5 27.6 27.55 0 199 62 8 8.1 8.05 0 388 199 36.8 6.2 6.4 6.3 22 22.1 22.05 0 198 46 6.6 6.3 6.45 27.2 27.5 27.35 0 184 62 7.1 7.3 7.2 0 351 200 33.5 5.7 5.7 5.7 23.5 23.9 23.7 0 103 42 6.2 6.5 6.35 22.1 22.5 22.3 0 193 54 6.3 6.4 6.35 

30 455 81 40.2 7.1 7.3 7.2 24.9 25 24.95 0 186 51 6.6 6.8 6.7 28.1 28.5 28.3 30 68 55 6.1 6.2 6.15 
B-1 

30 449 82 38.2 6.1 6 6.05 25.5 25.9 25.7 30 18 51 5.8 6 5.9 29.9 30 29.95 30 95 52 7.9 8 7.95 30 444 83 40.5 6.6 6.7 6.65 25.2 25 25.1 30 93 46 6.2 6.5 6.35 26 26.1 26.05 30 91 66 7.3 7.4 7.35 30 452 84 33.4 7.1 7.5 7.3 19.9 19.7 19.8 30 92 52 7.3 7.4 7.35 24.6 24.8 24.7 30 93 60 7.1 7.3 7.2 30 456 85 31.7 6.3 6.5 6.4 20 20.1 20.05 30 91 52 7.3 7.3 7.3 28.2 28.4 28.3 30 92 64 8 8.4 8.2 30 446 86 34 5.9 6.2 6.05 21.1 21.3 21.2 30 83 52 7 7.2 7.1 25.6 25.9 25.75 30 87 71 7.9 8.1 8 30 448 87 42.3 7.5 7.6 7.55 24.7 25 24.85 30 96 49 6.2 6.2 6.2 28.8 28.9 28.85 30 84 58 7.5 7.5 7.5 30 451 89 31.6 5.9 5.8 5.85 22.8 23 22.9 30 94 41 6.2 6 6.1 26.2 26.4 26.3 30 98 58 6.7 6.5 6.6 30 459 90 35.7 5.8 6 5.9 24.4 24.7 24.55 30 95 40 6.3 6.8 6.55 26.1 26.5 26.3 30 90 62 7.9 8.1 8 30 454 99 34.5 6.4 6.6 6.5 24.4 24.5 24.45 30 86 46 6.2 6.3 6.25 25.8 26.2 26 30 85 57 7 7.3 7.15 60 365 73 38.7 6.8 7.1 6.95 23.5 23.2 23.35 30 82 50 7 7.4 7.2 27.8 27.5 27.65 60 80 65 7.7 8 7.85 60 370 81 34.2 7.1 7.3 7.2 24.5 24.6 24.55 30 88 46 6.2 6.3 6.25 26.1 26.5 26.3 60 96 44 7.1 7.5 7.3 60 369 82 39.8 7 7.1 7.05 24.5 24.6 24.55 60 98 48 7.3 7.2 7.25 31.1 31.3 31.2 60 91 68 7.5 7.6 7.55 60 380 84 36.6 7.2 7.3 7.25 22.5 22.5 22.5 60 89 51 6.6 6 6.3 29.3 29.5 29.4 60 96 59 8.3 8.1 8.2 60 376 85 33.3 5.7 5.9 5.8 24.6 24.6 24.6 60 82 51 6.8 6.5 6.65 28.8 29 28.9 60 78 57 6.3 6.6 6.45 60 372 86 37.5 6 6.3 6.15 21.3 21.6 21.45 60 88 54 6.5 6.9 6.7 29.6 30 29.8 60 79 60 7.4 7.7 7.55 60 374 87 37.3 5.2 5.3 5.25 24.1 24.3 24.2 60 87 48 7.1 7.3 7.2 25.7 26.1 25.9 60 76 64 7.8 8 7.9 60 320 88 41.6 7.1 7.4 7.25 26.2 26.5 26.35 60 86 50 7 7.4 7.2 25.7 25.8 25.75 60 77 50 7.7 7.8 7.75 60 366 89 39.7 6.4 6.3 6.35 25.7 25.9 25.8 60 73 50 6.6 6.8 6.7 30 29.8 29.9 60 75 69 8.8 8.9 8.85 60 196 90 35.7 6.3 6.3 6.3 23.8 23.9 23.85 60 90 46 6.8 7.1 6.95 29.5 29.4 29.45 60 95 60 7.8 7.9 7.85 

125 392 73 37.2 6.3 6.4 6.35 21.9 22.1 22 60 74 40 6.3 6 6.15 25.4 25.8 25.6 125 94 62 7.2 7.3 7.25 125 396 74 40 6.5 6.9 6.7 23.3 23.6 23.45 60 81 44 6.7 7 6.85 27.5 26.9 27.2 125 81 58 7.9 7.9 7.9 125 382 75 40.9 6.8 7.1 6.95 22.2 22.4 22.3 125 90 46 7.4 7 7.2 27.7 28 27.85 125 96 52 7.5 7.8 7.65 125 398 76 36.4 6.3 6.5 6.4 24.2 24.3 24.25 125 91 49 6.9 6.9 6.9 28.8 28.5 28.65 125 82 66 7.3 7.5 7.4 125 345 77 33.6 6.1 6.5 6.3 21.7 21.9 21.8 125 75 51 6 6.3 6.15 28.4 28.2 28.3 125 99 62 7.5 7.6 7.55 125 393 78 35 6.1 6.3 6.2 23.5 23.5 23.5 125 74 52 7.1 7.1 7.1 25.8 26.1 25.95 125 95 63 6.9 7 6.95 125 391 79 36.2 7.8 4.3 6.05 23.6 23.8 23.7 125 76 47 6.1 6.4 6.25 27.4 27.5 27.45 125 100 59 7 7.3 7.15 125 375 80 35.2 7 7.2 7.1 22.2 22.6 22.4 125 77 45 7.6 7.4 7.5 27.6 27.6 27.6 125 98 57 7.3 7.3 7.3 125 387 90 36 6.4 6.6 6.5 25.5 25.8 25.65 125 83 50 6.8 6.9 6.85 26.6 26.8 26.7 125 97 65 6.8 6.9 6.85 125 383 91 37.7 2 6.3 4.15 22.2 22.5 22.35 125 80 46 6 6.1 6.05 24.2 24.5 24.35 125 90 59 7 7.1 7.05 250 415 81 42.2 6.1 6.3 6.2 22.2 22.5 22.35 125 92 41 7.1 7.2 7.15 26.3 25.9 26.1 250 87 65 7.9 8 7.95 250 419 82 40.2 7.1 7 7.05 23.2 23.3 23.25 125 93 50 8.1 8.3 8.2 28.2 28.4 28.3 250 88 74 7.8 8 7.9 250 427 83 33.7 7.1 7.3 7.2 24.1 24.3 24.2 250 30 35 7.1 7.4 7.25 22.1 22 22.05 250 89 61 7.9 8 7.95 250 414 84 36.8 6.9 7 6.95 21.2 21.5 21.35 250 78 45 6.9 6.7 6.8 28.2 28.6 28.4 250 90 48 5.6 5.6 5.6 250 125 85 30.1 6.3 6.7 6.5 21.2 21.4 21.3 250 89 49 7.2 7.3 7.25 24.2 24.4 24.3 250 74 49 7.1 7.3 7.2 
B-1 

250 413 85 46 6.7 6.9 6.8 25.5 25.2 25.35 250 73 35 5.2 5.4 5.3 23.2 23.6 23.4 250 76 60 6.9 7.1 7 250 429 87 37.5 6 6.2 6.1 25.5 25.2 25.35 250 84 48 7 7.1 7.05 26.3 26.5 26.4 250 75 66 7.8 8 7.9 250 422 89 36.8 6.8 6.9 6.85 21.9 21.8 21.85 250 83 44 6.5 6.7 6.6 24.5 25.3 24.9 250 73 45 6.7 6.8 6.75 250 428 90 23.8 5.4 5.9 5.65 17 17.6 17.3 250 81 54 6.4 6.7 6.55 27.7 28 27.85 250 78 59 7.9 8 7.95 250 426 91 32.8 6.4 6 6.2 21.9 21.4 21.65 250 85 41 5.9 6.2 6.05 25 25.1 25.05 250 91 57 7.8 7.9 7.85 

 250 87 49 6.4 6 6.2 26.6 26.3 26.45  250 82 53 7 7.1 7.05 27.1 26.8 26.95  250 88 60 6.5 6.5 6.5 30.6 30.8 30.7 

F2 growth measurements 

12/4 12/7 12/10 

DOSE ID BW tarsus 1 tarsus 2 avg tarsus culmen 1 culmen 2 avg culmen SEX DOSE ID BW tarsus 1 tarsus 2 avg tarsus culmen 1 culmen 2 avg culmen SEX DOSE ID BW tarsus 1 tarsus 2 avg tarsu

0 193 73 9 9.2 9.1 30.3 30.5 30.4 F 0 181 90 8.7 9 8.85 33.7 33.8 33.75 F 0 200 82 8.4 8.4 8.4 0 198 69 6.6 6.7 6.65 32.8 33 32.9 F 0 183 77 8.1 8.5 8.3 31.3 30.9 31.1 F 0 181 91 7.9 8 7.95 0 181 80 8.2 8.3 8.25 33.1 33 33.05 F 0 197 80 7.4 7.7 7.55 28.4 28.6 28.5 F 0 199 83 7.5 8 7.75 0 183 69 6.8 6.9 6.85 30.6 30.8 30.7 F 0 199 79 6.7 6.9 6.8 32 32.4 32.2 F 0 183 81 8.1 8.3 8.2 0 184 76 7.4 7.4 7.4 30.1 30.5 30.3 F 0 200 75 8.4 8.7 8.55 29.4 29.8 29.6 F 0 197 84 6 6.3 6.15 0 196 61 6.8 6.8 6.8 29.6 29.8 29.7 F 30 68 80 8.4 8.7 8.55 33.6 32.9 33.25 F 30 86 83 8 8 8 0 200 68 7.1 7.2 7.15 30.7 21 25.85 F 30 84 84 8.2 8.6 8.4 32.8 32.9 32.85 F 30 85 87 6.9 7.1 7 0 197 71 7 7.2 7.1 29.9 29.7 29.8 F 30 85 83 8 8.2 8.1 31.7 32 31.85 F 30 68 81 8.5 8.7 8.6 0 182 60 8.1 8.3 8.2 28.9 29 28.95 F 30 86 80 7.5 7.7 7.6 30.6 30.4 30.5 F 30 87 101 7.6 7.7 7.65 0 192 76 6.8 7 6.9 31.6 31.8 31.7 F 30 87 97 7.7 7.8 7.75 33.6 34 33.8 F 60 91 87 8 8.2 8.1 

30 86 72 7.1 7.3 7.2 32.3 32.4 32.35 F 60 75 89 9 9.4 9.2 34.6 34.9 34.75 F 60 92 84 7.9 8.1 8 30 89 68 7.7 7.5 7.6 31.2 31.1 31.15 F 60 77 73 8.2 8.4 8.3 33.8 33.9 33.85 F 60 75 94 8.3 8 8.15 30 91 79 8.4 8.5 8.45 32.7 33 32.85 F 60 78 82 8.1 8.4 8.25 33.7 33.9 33.8 F 60 76 88 8 8.1 8.05 30 84 73 8 8.4 8.2 31.6 31.5 31.55 F 60 92 83 7.6 7.8 7.7 27.3 27.5 27.4 F 60 93 76 7.1 7.2 7.15 30 95 63 7.1 7.4 7.25 30.6 31 30.8 F 60 93 74 7.7 7.9 7.8 29 29.4 29.2 F 60 94 82 8.6 8.7 8.65 30 94 63 7.8 8 7.9 30.5 30.7 30.6 F 60 94 79 8.5 8.8 8.65 33.1 33.2 33.15 F 125 74 86 7.7 7.5 7.6 30 93 71 8.1 8.3 8.2 30 30.8 30.4 F 125 74 85 7.4 7.5 7.45 34.9 34.5 34.7 F 250 78 91 7.4 7.5 7.45 30 87 86 7.2 7.5 7.35 32.5 32.4 32.45 F 250 78 82 7.8 7.9 7.85 32.2 32.6 32.4 F 250 74 74 7.6 7.8 7.7 30 85 71 7.2 7.4 7.3 28.7 28.9 28.8 F 250 88 95 7.3 7.5 7.4 33.3 33.3 33.3 F 250 88 97 8.5 8.9 8.7 30 90 77 8.1 8.3 8.2 33.8 33.9 33.85 F 250 89 83 7.4 7.5 7.45 32.7 33 32.85 M 0 196 73 7.4 7.6 7.5 60 78 69 7.1 7.2 7.15 32 32.3 32.15 M 0 182 69 7.7 7.5 7.6 32.5 32.7 32.6 M 0 184 87 8.5 8.6 8.55 60 77 62 7.2 7.2 7.2 32.6 32.3 32.45 M 0 196 71 8.2 8.3 8.25 27.3 27.4 27.35 M 0 192 86 7.7 7.8 7.75 
B-1 

60 76 77 7.5 7.8 7.65 31.8 32.2 32 M 0 198 77 7.9 8.2 8.05 30 30.1 30.05 M 0 182 72 7.5 7.6 7.55 60 92 75 7.1 7.1 7.1 32.2 31.5 31.85 M 30 91 86 8.4 8.5 8.45 31.7 32 31.85 M 0 198 79 7.6 7.8 7.7 60 96 57 7.5 7.8 7.65 29.7 30 29.85 M 30 92 70 7.9 8.1 8 29.8 29.7 29.75 M 30 94 74 7.3 7.5 7.4 60 94 70 8.6 8.6 8.6 31.8 32.2 32 M 30 93 82 7.4 7.5 7.45 32.2 32.3 32.25 M 30 90 89 7.8 8 7.9 60 75 83 7.8 7.9 7.85 32.7 32.9 32.8 M 30 95 71 7.1 7.2 7.15 29.2 29.2 29.2 M 30 92 78 7.5 7.4 7.45 60 79 73 7.4 7.4 7.4 30.9 31.1 31 M 60 79 80 8.1 8.5 8.3 32.8 32.9 32.85 M 30 95 74 7.3 7.4 7.35 60 95 74 7 7.2 7.1 30.7 30.8 30.75 M 60 90 85 6.9 7 6.95 31.2 31.2 31.2 M 30 93 84 7.1 7.2 7.15 60 91 80 7.1 7.2 7.15 32.5 32.6 32.55 M 60 95 82 8.6 8.9 8.75 30.7 30.6 30.65 M 30 91 89 8.1 7.9 8 

125 95 76 6.9 6.7 6.8 31.2 31.2 31.2 M 60 96 65 7.3 7.5 7.4 32.2 31.9 32.05 M 60 96 69 8.2 8.4 8.3 125 96 63 7.1 7.2 7.15 31.4 31.6 31.5 M 125 81 81 7.8 8 7.9 32.1 32.2 32.15 M 60 80 87 6.8 6.9 6.85 125 99 73 7.8 7.9 7.85 32.3 32 32.15 M 125 82 90 7.8 7.8 7.8 33.6 33.7 33.65 M 60 95 83 7.4 7.9 7.65 125 81 72 7.5 7.6 7.55 31.7 32 31.85 M 125 94 84 8.2 8.5 8.35 28.9 29.2 29.05 M 60 79 82 8.2 8.3 8.25 125 98 68 7.2 7.4 7.3 31.1 31.5 31.3 M 125 95 84 7.3 7.4 7.35 33.7 33.9 33.8 M 125 81 86 8.3 8.5 8.4 125 97 77 7.7 7.8 7.75 32.5 32.3 32.4 M 125 96 72 8.4 8.4 8.4 31.5 31.8 31.65 M 125 96 75 8 8.3 8.15 125 82 80 7.1 7.5 7.3 32.6 32.9 32.75 M 125 97 88 8.1 8.1 8.1 34.4 34.3 34.35 M 125 97 88 7.9 8 7.95 125 100 72 7.1 7.3 7.2 32.7 32.9 32.8 M 125 98 80 8.4 8.7 8.55 34.6 34.7 34.65 M 125 100 85 7.1 7.2 7.15 125 94 74 7.1 7.2 7.15 29 29.2 29.1 M 125 99 80 7.2 7.5 7.35 32.6 32.7 32.65 M 125 82 94 7.4 7.6 7.5 125 74 76 7.8 7.8 7.8 32.8 33 32.9 M 125 100 81 7.5 7.5 7.5 30.8 30.9 30.85 M 125 98 82 7.6 7.6 7.6 250 16 71 6.8 6.8 6.8 30.3 30.6 30.45 M 250 73 64 7.5 7.7 7.6 33.5 33.6 33.55 M 125 99 81 7.1 7.2 7.15 250 89 77 6.6 6.8 6.7 32.8 32.8 32.8 M 250 75 91 8.4 8.7 8.55 33.2 33.5 33.35 M 125 95 84 8.2 8.3 8.25 250 73 57 6.9 7 6.95 29.3 29.5 29.4 M 250 76 78 6.8 7.2 7 33 33.1 33.05 M 125 94 85 8.2 8.5 8.35 250 74 63 7.5 7.6 7.55 29.5 29.9 29.7 M 250 77 96 7.7 8 7.85 30.1 30.4 30.25 M 250 75 92 7.9 8 7.95 250 88 87 8.3 8.5 8.4 33.3 33.4 33.35 M 250 81 88 8.2 8.5 8.35 31.5 32 31.75 M 250 91 81 7.8 7.9 7.85 250 87 76 7.3 7.4 7.35 33.2 32.9 33.05 M 250 90 73 8 8.2 8.1 32.6 32.7 32.65 M 250 81 90 7.7 7.7 7.7 250 77 86 8 8.3 8.15 31.7 31.6 31.65 250 16 78 8.5 8.6 8.55 31.2 31 31.1 M 250 77 99 7.5 7.5 7.5 250 76 79 6.7 7 6.85 32.1 32.2 32.15 30 94 73 8 8.4 8.2 32.4 32.5 32.45 M 250 87 89 7 7.1 7.05 250 90 62 6.6 6.9 6.75 30.6 30.7 30.65 0 192 84 7.4 7.5 7.45 33.7 33.8 33.75 M 250 73 67 7.3 7.4 7.35 250 75 79 7 7.4 7.2 30.1 30.5 30.3 0 184 85 7 7.2 7.1 30.9 31 30.95 M 250 76 80 7.7 7.8 7.75 250 81 79 6.4 6.5 6.45 32 32.3 32.15 250 78 72 7.2 7.5 7.35 32.3 32.4 32.35 

F2 bird weights 

 12/22 12/28 1/5 1/12 1/19 1/26 2/2 2/9 2/16 2/22 3/2 3/9 3/16 3/22 12/22 12/28 1/5 1/12 1/19 1/26 2/2 2
B-1 

SEX DOSE ID BW BW BW BW BW BW BW BW BW BW BW BW BW BW SEX DOSE ID BW BW BW BW BW BW BW BF 0 183 92 103 113 134 127 120 133 132 139 142 137 136 138 140 M 0 193 91 86 97 101 103 105 106 1F 0 199 97 91 121 131 125 113 127 139 139 141 130 131 143 141 M 0 195 89 96 100 102 101 105 105 1F 0 200 97 107 123 130 129 118 127 132 128 133 120 122 129 135 M 0 196 94 99 103 104 104 105 101 1F 0 194 96 110 127 136 131 135 135 129 140 110 134 134 139 137 M 0 198 94 89 93 98 91 99 102 F 0 179 115 122 109 131 130 136 134 142 140 147 145 144 144 136 M 0 184 100 104 106 108 111 111 111 1F 0 191 92 92 100 121 126 123 127 126 126 135 133 137 131 132 M 0 192 95 100 99 105 103 103 105 1F 0 188 96 113 116 124 133 129 131 133 133 137 134 133 135 137 M 0 177 91 102 110 115 116 119 120 1F 0 189 123 138 139 136 128 137 147 145 147 147 144 143 140 144 M 0 180 92 104 113 114 109 115 116 1F 0 313 102 109 115 121 130 148 145 133 144 139 138 137 133 133 M 0 304 95 99 102 105 108 106 109 1F 0 174 106 115 122 137 133 136 142 140 141 146 142 143 140 137 M 0 308 95 98 103 107 107 107 112 1F 0 314 96 106 127 138 140 145 147 142 143 141 132 132 147 148 M 0 322 95 99 104 95 98 102 106 1F 0 309 95 107 121 125 127 130 130 126 126 129 128 129 131 135 M 0 321 91 102 112 102 108 110 113 1F 0 186 101 94 133 145 144 144 146 147 147 153 146 144 152 156 M 0 301 100 102 107 109 109 109 114 1F 0 197 92 105 133 134 141 134 135 137 139 142 135 134 137 133 M 0 182 91 97 103 103 104 106 110 1F 0 178 104 101 140 141 143 141 134 142 147 144 135 135 147 138 M 0 185 104 102 107 109 108 108 108 1F 0 181 101 105 127 144 141 130 143 144 144 150 145 146 139 149 M 0 173 93 91 97 97 98 98 100 F 0 175 107 111 134 144 137 146 146 145 149 143 144 145 139 140 M 0 187 91 88 94 97 96 95 104 F 0 190 81 94 112 128 141 118 142 138 146 145 147 146 152 151 M 0 303 93 98 102 100 102 106 105 1F 0 323 106 129 144 142 138 142 138 144 149 152 152 152 153 153 M 0 310 90 96 100 103 104 106 107 1F 0 305 119 123 129 147 125 121 147 141 139 155 144 143 153 160 M 0 329 102 109 115 113 114 107 115 1F 30 81 126 131 129 135 135 132 130 140 140 141 141 140 138 133 M 30 94 96 97 103 108 105 107 110 1F 30 96 102 115 127 114 132 131 128 132 128 135 1282 129 134 134 M 30 93 103 104 106 108 106 109 108 1F 30 89 101 105 122 138 142 140 138 140 141 146 134 133 145 140 M 30 90 110 119 118 118 118 121 123 1F 30 85 110 114 132 132 138 139 138 127 139 143 138 139 139 143 M 30 80 110 114 118 122 123 129 130 1F 30 403 91 106 112 119 131 136 134 140 139 141 144 143 138 143 M 30 78 97 100 105 108 110 112 113 1F 30 405 97 119 121 131 128 127 134 137 143 144 140 140 135 142 M 30 76 96 95 101 103 105 109 109 1F 30 408 118 129 133 137 138 138 137 139 141 143 135 137 141 138 M 30 407 100 102 103 106 108 110 110 1F 30 427 108 133 134 140 143 142 143 146 141 144 140 141 141 144 M 30 402 100 102 105 107 106 106 112 1F 30 409 101 106 111 119 130 152 144 145 140 149 145 145 148 151 M 30 417 101 105 106 109 97 100 106 F 30 432 106 119 127 139 146 133 149 151 150 154 152 151 152 150 M 30 406 93 98 97 102 103 105 107 1F 30 84 105 122 122 132 123 135 132 132 136 137 127 128 131 136 M 30 92 100 110 115 118 123 120 123 1F 30 86 101 107 111 122 128 127 128 125 132 132 131 132 129 133 M 30 88 103 104 105 106 108 105 109 1F 30 82 107 116 124 134 131 132 140 139 133 141 140 141 143 140 M 30 95 93 95 100 102 118 105 105 1
B-1 

F 30 87 117 127 149 145 151 140 150 156 159 163 167 166 162 170 M 30 91 105 107 112 111 107 108 111 1F 30 401 104 111 122 126 128 148 148 150 149 154 150 149 140 139 M 30 77 102 105 115 116 122 119 121 1F 30 74 106 117 125 123 126 138 140 141 143 145 144 145 142 146 M 30 83 104 104 106 106 105 105 106 1F 30 73 107 112 118 125 135 137 138 138 141 133 143 143 142 143 M 30 404 106 120 120 123 108 126 126 1F 30 86 88 100 118 130 126 133 131 128 135 135 135 135 125 136 M 30 100 89 97 102 103 103 107 110 1F 30 75 119 124 132 138 138 136 141 139 139 146 144 143 144 147 M 30 99 98 102 104 107 103 108 109 1F 30 79 120 106 114 130 128 115 128 125 125 137 118 119 135 133 M 30 97 102 104 105 105 111 103 104 1F 60 75 114 128 149 149 150 151 147 159 151 150 157 155 159 159 M 60 90 98 100 103 106 104 108 107 1F 60 91 101 110 128 136 129 143 136 140 140 144 144 143 139 145 M 60 88 100 102 108 109 110 109 112 1F 60 89 113 120 126 135 140 149 144 150 143 147 144 145 142 147 M 60 205 85 88 90 94 95 97 98 1F 60 73 102 106 121 134 141 134 140 146 139 137 144 144 139 142 M 60 208 103 104 107 110 110 115 119 1F 60 85 101 108 126 144 127 135 148 140 147 148 135 137 143 129 M 60 81 94 99 100 99 101 101 103 1F 60 100 121 129 137 138 135 145 142 139 148 148 150 146 143 133 M 60 84 94 99 101 105 105 107 108 1F 60 94 97 120 124 123 129 127 132 124 123 135 116 117 131 118 M 60 80 105 107 111 114 113 109 108 1F 60 76 105 112 125 132 142 139 134 139 138 135 140 140 140 138 M 60 86 104 105 109 113 114 116 117 1F 60 78 98 104 124 134 109 108 138 139 143 142 134 133 133 142 M 60 96 93 97 104 103 104 106 105 1F 60 93 93 110 122 124 123 123 118 125 120 128 125 125 128 112 M 60 79 104 108 117 112 106 106 112 1F 60 82 95 103 116 122 127 126 122 131 127 123 120 121 130 128 M 60 203 98 103 109 109 113 108 118 1F 60 74 96 104 119 129 126 125 129 129 130 133 122 122 126 134 M 60 95 102 107 113 115 114 117 115 1F 60 92 110 117 127 132 125 135 137 135 143 139 140 140 142 131 M 60 99 112 112 112 112 115 115 121 1F 60 87 94 97 124 133 135 113 136 140 140 131 124 125 141 145 M 60 98 94 103 109 111 112 120 112 1F 60 202 89 93 115 136 122 138 144 139 140 145 136 135 145 146 M 60 201 85 92 99 101 102 109 101 1F 60 77 95 83 96 118 135 135 137 137 137 140 128 129 136 136 M 60 204 86 95 100 103 105 106 104 1F 60 222 97 96 112 127 137 144 143 143 142 152 150 151 150 151 M 60 209 104 104 101 107 110 113 111 1F 60 97 123 118 135 115 133 122 137 136 136 142 136 137 123 133 M 60 210 95 103 111 115 116 115 119 1F 60 215 98 104 133 131 131 135 136 136 139 134 140 140 137 140 M 60 219 95 106 104 107 108 107 108 1F 60 216 99 102 130 137 136 122 137 139 143 146 144 144 144 135 M 60 218 101 103 111 114 117 116 117 1F 125 88 93 109 119 133 137 140 146 150 149 145 141 139 142 152 M 125 75 101 103 107 106 106 112 112 1F 125 87 100 102 121 123 132 125 136 136 134 137 139 141 128 130 M 125 77 100 103 104 105 106 107 109 1F 125 89 99 115 128 133 124 133 135 137 123 123 131 130 136 138 M 125 95 107 107 110 116 114 115 121 1F 125 84 92 103 119 108 134 134 129 111 135 139 136 135 138 140 M 125 233 88 101 109 110 110 109 111 1F 125 207 105 119 136 122 145 148 149 152 153 153 150 150 149 156 M 125 100 101 102 102 107 110 110 109 1F 125 202 105 118 120 135 141 145 146 149 152 150 150 151 150 151 M 125 81 104 110 114 118 119 124 124 1F 125 259 104 113 123 134 133 137 135 131 132 142 126 127 140 140 M 125 98 105 106 107 111 113 113 114 1
B-1 

F 125 209 104 123 125 131 141 143 144 145 144 146 139 138 147 147 M 125 97 99 109 115 116 117 120 122 1F 125 258 103 106 116 124 140 142 133 144 144 132 129 129 145 149 M 125 208 96 99 102 106 118 107 110 1F 125 260 98 115 147 152 155 154 147 157 153 148 155 157 155 155 M 125 76 107 113 114 111 114 115 117 1F 125 74 103 106 109 117 134 137 136 134 126 141 135 134 137 143 M 125 235 103 112 114 114 114 115 115 1F 125 80 100 94 119 130 134 125 121 126 120 137 128 129 122 141 M 125 93 106 105 107 110 111 109 114 1F 125 83 115 118 132 135 145 159 154 149 146 155 151 152 157 157 M 125 82 112 110 109 108 114 113 116 1F 125 73 111 120 135 127 123 140 139 131 136 147 129 128 145 147 M 125 94 108 109 114 116 118 116 119 1F 125 206 105 116 134 128 130 132 139 141 139 143 140 141 135 142 M 125 90 100 102 107 106 109 111 112 1F 125 205 118 130 136 142 159 160 165 162 162 162 164 162 161 161 M 125 99 102 109 112 115 116 119 120 1F 125 234 106 113 121 136 134 129 137 142 141 145 143 141 141 136 M 125 78 101 109 113 117 116 115 116 1F 125 201 111 117 132 126 124 126 133 135 136 116 131 132 135 134 M 125 85 103 108 111 112 112 115 115 1F 125 239 102 111 115 122 127 140 133 141 144 147 145 146 140 146 M 125 204 109 113 110 112 113 112 113 1F 125 237 95 104 117 134 130 137 136 128 138 139 130 130 140 135 M 125 79 104 108 112 117 117 114 113 1F 250 89 118 136 140 140 145 138 148 142 150 148 150 147 146 149 M 250 84 97 104 109 113 115 114 116 1F 250 82 119 112 121 124 129 130 135 139 136 141 138 137 140 140 M 250 83 106 101 103 103 108 109 113 1F 250 79 107 113 139 130 126 137 134 145 141 148 144 143 148 139 M 250 87 104 113 117 118 121 121 122 1F 250 92 102 104 116 113 120 119 132 139 140 132 109 109 114 120 M 250 91 101 106 106 105 108 110 114 1F 250 94 102 107 116 125 138 135 141 140 134 140 141 142 142 136 M 250 77 121 125 101 130 130 133 135 1F 250 228 101 107 129 132 134 132 136 122 136 137 138 137 139 128 M 250 76 94 94 94 97 98 99 100 1F 250 188 95 98 102 105 105 111 121 117 128 134 138 140 137 139 M 250 98 100 98 98 93 98 102 103 1F 250 184 105 126 128 123 132 129 125 128 133 139 134 135 138 133 M 250 100 99 103 104 111 110 120 119 1F 250 97 112 114 134 139 134 138 113 140 142 141 146 145 144 145 M 250 95 113 98 104 106 110 109 110 1F 250 74 99 116 141 132 132 146 141 152 140 154 148 149 148 154 M 250 99 100 103 102 103 103 105 108 1F 250 78 118 123 136 140 139 145 140 147 145 139 140 142 149 148 M 250 231 94 98 105 104 105 106 108 1F 250 88 122 133 141 149 149 142 147 141 150 147 148 147 139 151 M 250 189 95 99 103 108 109 110 112 1F 250 96 118 125 132 144 133 147 147 142 151 151 152 151 152 151 M 250 230 95 103 106 106 108 107 107 1F 250 93 117 124 135 134 140 140 145 146 142 153 142 144 148 146 M 250 90 99 101 107 109 108 111 112 1F 250 225 110 116 121 125 137 143 148 145 143 150 145 146 140 144 M 250 75 103 107 103 105 107 107 111 1F 250 221 109 118 128 140 147 143 149 150 145 156 153 153 150 157 M 250 81 106 109 111 112 112 111 114 1F 250 223 102 107 109 113 118 120 124 128 131 136 136 135 138 138 M 250 85 106 112 110 117 119 120 123 1F 250 183 105 107 107 104 104 105 107 108 110 111 110 111 111 112 M 250 80 92 93 91 93 93 94 95 F 250 182 100 114 123 128 129 129 129 130 132 132 132 133 133 133 M 250 181 99 104 107 111 110 114 112 1F 250 187 100 114 117 121 140 138 155 152 156 151 155 155 147 148 M 250 190 96 100 101 100 100 101 104 1
B-1 

F2 feed consumption 

 12/21 12/28 1/4 1/11 1/18 1/25 1/29 2/1 2/5 2/8 2/12 2/15 DOSE SEX ID Initial Initial Final Initial Final Initial Final Initial Final Initial Final Initial Final Initial Final Initial Final Initial Final Initial Final Initial 

0 M 193 389 324 326 322 433 306 425 320 426 314 428 313 426 305 431 362 429 0 M 195 386 319 431 324 432 317 426 325 427 329 419 312 426 326 444 378 427 0 M 196 386 308 429 319 437 330 433 322 422 327 421 323 430 325 442 365 412 0 M 198 388 325 435 325 431 305 421 320 429 302 412 322 445 355 439 306 432 359 428 0 M 194 428 324 434 311 411 334 414 313 433 317 402 283 430 305 442 363 437 0 M 192 438 322 432 331 433 310 419 334 418 331 432 322 421 319 423 354 429 0 M 177 425 321 432 310 430 303 418 308 412 313 413 286 428 324 441 355 424 0 M 180 428 320 435 291 435 293 424 297 428 306 418 279 431 303 439 338 418 0 M 304 427 309 435 318 427 310 421 317 416 338 421 313 426 282 436 363 424 0 M 308 421 315 427 324 434 311 419 326 426 319 424 319 423 322 437 366 416 0 M 322 410 316 431 322 430 339 406 328 404 310 425 321 425 297 425 369 424 0 M 321 404 311 426 285 432 318 421 290 411 290 428 298 438 312 430 350 445 0 M 301 436 312 424 308 440 317 438 307 425 314 407 295 438 310 433 352 443 0 M 182 422 282 437 305 445 325 439 308 420 318 407 301 439 376 426 298 424 348 452 0 M 185 413 317 434 306 441 305 439 307 404 313 433 311 439 320 438 364 409 0 M 173 431 317 435 282 436 272 436 284 425 303 423 285 435 284 434 352 419 0 M 187 411 330 427 308 434 291 434 309 433 321 435 311 435 301 441 361 458 0 M 303 412 317 424 316 443 331 441 311 418 330 421 320 427 323 440 368 433 0 M 310 434 325 434 309 440 315 426 312 436 326 433 310 430 318 424 352 435 0 M 329 400 320 431 303 440 312 436 306 430 331 426 331 432 307 441 340 420 0 F 183 406 316 431 309 433 301 427 346 421 298 418 275 439 326 455 369 415 315 452 305 0 F 199 424 318 433 300 428 276 406 335 419 300 434 278 436 341 435 353 428 353 431 281 0 F 200 409 335 425 275 432 251 425 276 427 268 404 307 431 331 438 316 449 356 421 326 435 307 0 F 194 405 294 426 336 424 268 433 332 442 231 412 242 439 343 449 333 422 336 421 307 448 243 0 F 179 408 308 434 295 426 309 429 299 326 264 421 310 437 337 429 310 445 328 433 323 424 311 0 F 191 409 332 434 325 426 315 432 330 426 265 411 315 429 352 447 335 440 343 432 343 419 314 0 F 188 395 319 436 381 425 318 428 387 427 305 423 272 413 326 431 279 450 337 418 301 440 271 0 F 189 493 330 423 344 432 325 433 342 430 344 430 298 421 335 437 355 430 332 430 294 0 F 313 427 301 425 339 432 286 432 335 431 273 405 302 418 329 429 313 443 342 428 320 419 303 0 F 174 413 295 430 304 431 301 430 350 442 337 450 296 429 311 432 285 437 319 417 285 443 296 0 F 314 433 332 428 286 425 296 433 291 423 310 419 298 424 334 439 316 451 318 419 323 417 297 
B-1 

0 F 309 348 316 436 316 421 295 424 320 434 313 424 278 436 316 441 341 427 320 426 293 0 F 186 410 363 435 257 423 241 429 351 446 293 408 311 430 252 437 356 420 298 442 303 0 F 197 432 331 442 280 426 271 433 338 430 332 424 295 421 330 438 312 437 339 425 306 438 290 0 F 178 421 312 431 404 419 281 426 356 441 339 415 285 438 337 438 322 429 342 431 305 442 284 0 F 181 440 315 433 331 399 269 437 327 445 292 429 285 433 327 445 330 435 327 430 331 431 281 0 F 175 428 319 434 287 413 268 432 281 428 263 402 308 433 343 434 313 437 338 419 336 439 302 0 F 190 408 346 430 307 421 293 437 331 432 302 423 280 434 337 431 296 436 357 432 321 410 281 0 F 323 405 320 432 344 422 282 418 357 428 302 413 323 432 369 440 337 445 350 429 305 422 323 0 F 305 398 332 427 304 423 268 414 399 434 294 429 277 437 317 436 365 419 301 415 305 

30 M 94 415 323 422 301 427 313 433 295 431 321 422 316 424 315 432 373 431 30 M 93 423 320 433 316 432 325 438 312 436 304 427 301 415 327 443 354 442 30 M 90 401 325 422 305 423 318 438 300 435 300 422 307 436 326 435 352 436 30 M 80 411 315 431 315 421 295 420 302 422 302 418 289 423 291 436 346 436 30 M 78 404 314 432 316 435 331 423 303 423 320 411 303 434 333 435 359 437 30 M 76 403 305 428 316 420 299 429 303 413 301 421 313 427 313 437 367 429 30 M 407 402 317 432 325 429 297 427 319 405 278 407 271 429 331 431 344 452 30 M 402 393 308 434 314 429 301 420 310 406 321 415 292 435 310 432 353 437 30 M 417 382 319 425 327 443 325 420 322 417 343 404 316 425 337 437 366 429 30 M 406 395 315 437 314 428 305 430 309 422 304 429 309 432 308 438 359 429 30 M 92 446 311 421 302 432 276 437 295 423 299 420 299 426 305 431 357 415 30 M 88 422 289 430 326 433 346 433 319 425 332 435 305 437 336 445 361 416 30 M 95 433 316 428 306 442 311 433 302 423 320 419 315 439 311 434 344 411 30 M 91 429 317 428 322 429 315 433 317 417 335 422 320 426 330 435 372 440 30 M 77 430 330 430 292 439 301 424 290 429 300 423 317 433 314 437 356 438 30 M 83 425 316 432 321 430 310 420 317 433 344 432 299 423 332 438 357 434 30 M 404 421 321 421 306 437 296 427 299 450 307 418 302 425 292 430 345 413 30 M 100 431 325 432 315 434 325 436 298 413 313 408 305 433 325 431 366 431 30 M 99 441 324 431 312 437 317 425 310 410 326 421 324 435 328 435 374 433 30 M 97 412 332 433 324 427 330 442 318 428 337 418 317 428 314 439 361 437 30 F 81 438 305 432 294 426 242 420 327 413 307 421 297 420 309 432 312 436 320 426 305 436 296 30 F 96 424 335 434 345 428 310 436 338 420 325 434 304 426 326 445 367 408 312 437 292 30 F 89 424 309 432 338 406 306 429 330 430 283 421 293 420 320 433 327 441 339 428 323 428 294 30 F 85 415 315 423 314 418 297 423 362 442 310 423 299 432 323 442 331 417 320 433 335 435 298 30 F 403 424 316 433 266 431 312 423 266 424 285 409 305 421 313 436 326 433 299 444 303 
B-1 

30 F 405 418 327 432 313 427 297 416 310 437 284 414 296 418 323 430 323 427 338 421 329 444 298 30 F 408 420 317 428 329 426 308 434 322 426 313 424 301 423 324 429 314 420 317 431 305 431 305 30 F 427 415 330 428 303 426 304 420 301 448 290 429 294 428 334 424 365 427 339 441 308 30 F 409 411 328 430 301 421 301 425 298 435 281 410 304 420 343 439 347 433 357 422 319 449 312 30 F 432 423 317 428 293 420 318 432 291 437 322 431 318 430 351 445 345 443 352 435 342 447 315 30 F 84 428 318 436 276 428 293 422 275 436 300 423 301 431 345 435 350 325 343 429 314 30 F 86 410 334 425 315 424 292 427 310 429 290 422 292 416 328 438 356 333 349 413 310 30 F 82 419 299 424 312 417 321 423 311 425 259 420 314 423 356 443 342 444 366 329 328 421 313 30 F 87 491 308 425 251 403 272 415 259 428 284 426 287 430 330 448 320 434 319 429 309 441 288 30 F 401 411 331 434 297 419 283 416 287 437 293 423 309 414 328 443 327 421 337 332 291 450 310 30 F 74 420 302 432 308 431 288 416 301 438 273 422 321 425 346 449 333 427 327 422 304 443 322 30 F 73 411 296 422 314 421 315 417 313 427 319 416 310 421 344 432 339 431 319 424 305 433 312 30 F 98 413 331 431 313 414 299 427 310 422 270 424 310 424 353 446 346 429 340 429 293 444 310 30 F 75 407 333 429 330 431 295 419 339 418 321 430 313 428 336 438 316 442 322 425 319 428 312 30 F 79 413 317 435 363 419 283 418 361 433 305 416 319 423 344 442 345 433 365 409 338 429 320 60 M 90 435 340 425 274 424 265 410 276 422 303 422 307 431 325 431 339 428 60 M 88 423 325 434 331 425 305 427 326 434 308 415 305 430 309 427 365 416 60 M 205 419 332 423 309 434 300 426 301 440 319 426 302 425 318 436 347 446 60 M 208 417 327 432 319 422 301 426 312 432 300 412 303 417 309 427 341 433 60 M 81 408 341 435 323 432 326 468 318 427 304 421 287 424 282 435 356 415 60 M 84 422 340 424 304 435 300 433 297 422 320 430 291 434 329 441 347 434 60 M 80 424 332 423 308 431 305 421 308 405 317 410 287 415 306 432 363 415 60 M 86 413 340 438 313 429 298 423 310 423 314 426 285 431 281 428 327 428 60 M 96 421 319 435 310 448 310 429 305 432 320 428 312 424 302 438 350 415 60 M 79 423 322 439 302 437 316 418 198 419 311 427 301 428 319 431 356 433 60 M 203 414 340 426 301 438 307 423 302 432 301 420 268 435 308 431 342 414 60 M 95 408 335 429 307 454 310 426 301 421 336 426 320 425 326 434 366 439 60 M 99 443 325 431 319 430 316 423 311 427 302 419 290 420 308 431 358 414 60 M 98 413 320 434 320 435 324 419 315 426 341 423 308 439 319 423 353 432 60 M 201 407 321 425 299 434 324 417 286 425 322 418 317 423 318 436 367 428 60 M 204 412 336 432 310 423 306 423 299 423 323 423 320 445 309 442 368 420 60 M 209 417 342 425 315 440 312 426 309 440 301 418 298 438 282 429 355 417 60 M 210 423 331 433 296 419 286 419 297 442 304 425 298 424 304 436 352 446 60 M 219 430 324 430 317 427 317 430 315 429 333 426 330 440 328 432 368 412 
B-1 

60 M 218 422 318 425 287 433 279 424 347 430 269 430 275 435 257 436 281 425 322 418 60 F 75 420 303 426 286 417 275 424 341 432 299 413 267 432 313 425 328 435 322 412 263 443 268 60 F 91 415 337 428 286 420 308 420 290 434 283 410 296 418 341 434 318 434 344 406 308 446 295 60 F 89 422 341 432 301 428 306 416 287 432 311 425 306 437 305 436 344 427 334 415 272 60 F 73 410 360 426 304 412 255 413 353 430 328 430 267 424 324 438 336 429 324 423 306 442 365 60 F 85 401 349 433 284 426 293 425 279 440 307 419 277 430 336 429 360 425 339 414 255 60 F 100 387 299 424 300 424 299 422 294 436 267 429 251 431 327 437 317 430 334 423 313 423 250 60 F 94 399 340 434 358 422 327 430 349 439 258 427 305 424 342 432 321 420 322 404 308 420 304 60 F 76 388 341 432 340 420 282 410 334 435 287 417 290 429 279 427 267 438 328 411 309 424 292 60 F 78 381 336 430 352 419 248 403 349 429 352 432 292 450 340 437 337 408 305 437 310 60 F 93 438 338 427 366 434 302 427 361 434 287 434 328 428 352 444 345 424 354 431 335 428 326 60 F 82 426 341 432 304 422 308 421 361 435 267 429 297 443 364 445 348 440 339 413 346 418 299 60 F 74 428 340 433 305 437 299 425 299 428 288 431 313 436 357 440 325 421 353 405 308 445 310 60 F 92 417 302 431 359 422 281 413 349 425 285 434 277 442 349 443 321 432 324 422 328 407 275 60 F 87 401 339 436 305 417 292 424 300 423 314 431 295 440 332 429 326 429 325 449 283 60 F 202 431 342 434 295 404 302 423 352 424 279 419 298 431 348 437 331 426 336 425 316 448 296 60 F 77 407 359 436 339 418 281 410 364 435 349 419 282 443 344 421 358 423 329 430 295 60 F 222 410 360 437 317 415 299 421 311 425 338 430 339 436 309 433 323 405 319 418 292 60 F 97 399 298 433 359 426 313 422 342 438 267 412 299 425 330 430 317 423 341 410 313 403 296 60 F 215 394 340 427 321 419 281 427 440 258 429 311 427 344 447 335 435 343 429 313 450 310 60 F 216 400 340 432 367 415 300 413 317 421 283 418 311 429 338 447 324 440 323 410 308 444 302 

125 M 75 415 322 436 304 429 312 423 300 419 306 424 319 428 324 442 371 421 125 M 77 394 309 425 308 438 296 428 302 421 291 418 295 423 286 427 329 418 125 M 95 393 320 438 315 424 291 425 313 425 318 418 309 428 340 412 340 426 125 M 233 393 301 424 306 428 308 422 299 427 328 427 313 427 333 429 358 428 125 M 100 392 321 436 319 437 303 424 313 412 298 416 306 419 302 435 356 436 125 M 81 399 332 435 308 437 315 431 301 414 289 415 282 445 334 430 351 438 125 M 98 405 316 439 267 438 278 412 270 425 295 411 282 439 319 427 342 412 125 M 97 402 317 435 307 436 297 432 302 428 303 418 291 436 310 437 361 423 125 M 208 386 312 438 329 429 300 418 321 426 324 425 319 427 309 435 371 431 125 M 76 395 297 434 286 451 310 425 283 421 302 417 321 448 355 444 296 438 327 444 125 M 235 422 285 430 273 444 299 426 243 430 299 418 294 432 296 443 358 430 125 M 93 431 320 428 319 428 304 421 315 422 279 421 299 426 310 426 343 411 125 M 82 427 337 432 326 430 335 430 325 437 307 425 298 427 309 411 358 418 
B-1 

125 M 94 429 317 436 299 437 322 427 297 435 317 427 311 426 321 430 360 437 125 M 90 424 321 425 310 441 310 415 305 430 315 428 308 440 332 434 356 412 125 M 99 419 332 432 303 426 306 423 304 440 295 421 300 443 305 431 335 414 125 M 78 432 327 435 317 437 321 430 308 438 311 412 315 430 331 428 369 416 125 M 85 444 318 437 304 424 301 430 298 436 289 418 290 428 338 438 336 429 125 M 204 415 325 430 338 442 320 430 331 437 307 414 300 419 289 334 349 430 125 M 79 431 336 434 322 430 273 413 315 431 331 424 337 438 320 437 361 425 125 F 88 425 312 435 306 436 296 413 304 433 316 426 282 428 323 424 342 410 303 446 295 125 F 87 423 325 426 296 425 311 421 366 437 303 431 273 421 318 426 343 417 320 432 293 125 F 89 417 327 437 312 420 326 425 299 446 299 423 320 433 352 437 346 428 356 424 332 403 321 125 F 84 422 327 424 307 413 281 414 302 445 307 436 289 428 331 436 408 430 341 440 355 125 F 207 419 338 435 277 422 284 414 365 441 312 426 285 430 319 442 283 441 333 438 307 414 283 125 F 202 421 312 436 330 417 262 420 351 436 307 426 313 434 362 335 335 440 341 417 304 447 310 125 F 259 412 316 439 310 407 306 413 301 434 288 415 314 419 352 442 355 442 361 429 330 454 312 125 F 209 414 317 432 355 405 266 420 349 443 281 422 305 418 320 438 339 430 350 426 329 414 300 125 F 258 423 322 435 295 413 309 415 287 440 291 422 314 425 347 432 364 440 347 445 298 125 F 260 429 315 438 397 421 279 403 350 432 331 427 303 431 328 435 319 448 343 406 309 429 301 125 F 74 408 336 434 344 430 284 430 338 430 269 426 292 436 340 444 362 419 345 400 289 125 F 80 415 338 427 299 411 280 410 348 432 264 427 279 432 319 437 341 412 330 423 291 125 F 83 402 316 432 286 412 289 416 288 434 253 430 271 430 329 447 352 422 320 405 295 125 F 73 409 310 430 298 429 312 424 295 432 271 418 285 433 342 445 325 423 363 432 320 405 280 125 F 206 415 332 434 300 431 281 414 357 432 336 431 295 434 338 437 340 440 330 421 296 125 F 205 414 324 434 297 427 313 426 287 432 299 433 316 424 341 440 337 420 332 425 310 446 314 125 F 234 412 320 433 300 418 316 428 301 426 289 421 310 443 345 420 407 419 347 415 311 125 F 201 409 318 428 320 422 310 432 320 422 293 420 317 423 355 442 349 433 345 428 331 450 313 125 F 239 414 334 433 368 424 310 419 361 423 308 418 287 429 330 429 357 439 325 407 315 125 F 327 434 309 430 335 426 288 405 359 426 273 433 320 418 342 449 336 438 343 405 308 447 310 250 M 84 390 321 435 319 430 312 414 309 432 297 421 307 425 315 428 342 432 250 M 83 385 310 438 314 436 316 426 350 434 290 422 302 423 288 436 357 426 250 M 87 386 322 431 289 424 286 425 287 425 290 428 290 435 301 441 344 433 250 M 91 386 305 425 304 430 355 430 306 432 289 438 303 445 300 434 358 428 250 M 77 403 318 434 278 428 285 426 282 429 286 421 286 441 284 431 332 420 250 M 76 437 328 435 322 445 317 442 320 440 300 423 308 428 316 434 359 427 250 M 98 411 317 436 340 441 370 442 338 442 314 421 316 437 343 429 388 416 
B-1 

250 M 100 420 315 439 341 446 336 443 338 445 295 420 306 417 296 423 361 418 250 M 95 428 300 438 300 432 290 425 302 432 300 412 301 434 313 430 364 423 250 M 99 426 297 438 320 445 345 436 318 442 303 424 303 422 314 430 349 421 250 M 231 427 285 432 311 433 321 421 313 432 300 422 313 432 314 430 360 412 250 M 189 424 324 423 327 432 306 415 325 423 316 421 304 439 312 437 353 435 250 M 230 439 322 427 325 441 328 408 324 441 320 418 310 434 297 443 366 419 250 M 90 436 333 429 307 439 317 413 306 437 315 417 320 445 342 433 360 435 250 M 75 429 328 428 319 432 335 409 324 429 307 426 304 421 315 432 357 424 250 M 81 391 319 432 305 428 307 410 299 438 293 417 294 417 285 435 347 432 250 M 85 413 328 433 324 427 258 402 346 427 294 418 259 426 319 436 295 428 311 426 250 M 80 407 335 424 335 436 331 414 334 442 310 420 318 424 306 422 364 425 250 M 181 435 324 435 311 442 310 415 309 436 303 413 329 429 287 428 374 426 250 M 190 423 333 436 321 427 323 411 320 427 313 422 316 442 314 423 364 426 250 F 89 411 317 435 310 415 263 413 340 432 289 416 232 431 310 440 272 434 320 416 256 418 229 250 F 82 410 317 435 324 420 275 414 353 429 327 422 322 428 336 440 319 428 340 422 319 443 320 250 F 79 428 334 436 317 426 288 433 310 431 283 481 310 424 342 437 335 423 342 425 332 412 308 250 F 92 430 318 425 290 421 329 435 285 425 328 423 322 440 262 437 369 437 359 446 310 250 F 94 421 332 424 290 403 264 424 283 455 318 420 309 426 335 439 324 426 335 421 300 442 309 250 F 228 405 335 432 321 422 301 421 316 427 285 416 318 421 347 440 336 445 364 415 337 428 315 250 F 188 396 298 428 384 433 315 428 379 421 311 418 321 443 354 442 384 444 360 449 310 250 F 184 407 320 430 355 419 296 424 349 433 297 411 311 427 347 437 358 427 349 429 254 427 308 250 F 97 405 331 431 355 426 299 431 348 425 284 425 300 423 344 440 340 435 340 421 318 451 300 250 F 74 432 330 432 328 427 319 435 321 422 264 423 308 426 323 436 324 422 322 431 308 450 304 250 F 78 427 305 429 285 422 301 422 321 423 307 415 261 435 298 426 303 444 321 430 309 444 262 250 F 88 402 335 432 313 429 265 419 316 413 297 420 269 428 285 433 276 445 341 420 318 418 265 250 F 96 425 340 432 381 433 277 421 349 421 297 418 289 437 321 425 272 423 348 427 287 445 284 250 F 93 420 359 428 315 418 289 426 342 425 276 420 283 430 327 430 301 441 383 440 301 453 281 250 F 225 433 339 430 308 412 304 415 301 432 274 409 319 426 367 447 336 443 365 417 302 444 300 250 F 221 431 341 424 295 428 293 424 291 423 332 420 289 429 329 441 336 420 331 436 398 250 F 223 388 342 431 317 426 306 420 309 421 299 423 277 431 358 429 365 433 349 437 282 250 F 183 410 343 434 317 416 320 425 310 422 301 424 296 425 352 424 340 422 364 446 283 250 F 182 431 341 435 338 404 303 430 339 429 285 409 340 435 319 437 341 424 340 424 263 250 F 187 393 339 425 329 411 327 439 322 433 341 418 290 431 337 433 322 420 318 411 264 
B-1 

F2 feed consumption 

 2/22 2/26 3/1 3/5 3/8 3/12 3/15 3/19 3/22 3/26 DOSE SEX ID Initial Final Initial Final Initial Final Initial Final Initial Final Initial Final Initial Final Initial Final Initial Final Initial Final 

0 M 193 326 451 324 438 322 437 325 444 0 M 195 317 445 319 438 340 430 338 446 0 M 196 336 440 330 440 325 433 330 455 0 M 198 306 438 316 435 303 463 307 459 0 M 194 316 438 315 440 310 453 312 438 0 M 192 325 436 321 439 353 446 349 433 0 M 177 316 444 315 435 326 432 331 437 0 M 180 302 449 305 440 313 440 317 442 0 M 304 317 443 314 448 348 442 342 432 0 M 308 341 436 342 450 337 453 335 444 0 M 322 313 435 320 437 340 430 336 448 0 M 321 304 447 303 440 310 455 314 454 0 M 301 328 419 330 445 311 436 309 450 0 M 182 314 447 314 436 302 453 305 432 0 M 185 321 439 323 434 323 427 318 451 0 M 173 308 450 310 440 309 445 307 443 0 M 187 350 426 349 445 317 454 314 442 0 M 303 321 441 323 442 350 444 351 429 0 M 310 347 428 345 444 336 430 338 432 0 M 329 315 441 312 435 303 442 305 438 0 F 183 351 455 339 430 331 403 343 425 328 440 331 454 312 442 327 452 325 451 333 434 0 F 199 344 456 346 430 342 422 342 444 340 444 340 440 310 440 345 450 337 442 341 428 0 F 200 364 457 342 429 340 414 340 433 339 451 342 439 320 444 340 446 339 440 327 440 0 F 194 373 460 360 429 344 419 352 442 342 453 333 446 322 453 331 448 340 439 341 425 0 F 179 332 460 316 430 326 393 318 447 329 448 334 437 321 451 332 443 325 452 329 430 0 F 191 340 453 331 435 332 400 330 444 332 459 339 435 323 453 342 438 330 454 308 429 0 F 188 322 448 297 448 326 438 303 441 321 447 279 461 311 442 282 434 317 440 289 458 0 F 189 351 451 360 423 340 417 356 446 332 446 351 464 307 452 352 452 329 460 337 433 0 F 313 335 435 300 444 309 450 305 422 312 440 327 436 314 460 324 445 309 452 339 431 0 F 174 315 443 294 441 302 441 294 429 305 438 297 430 332 440 299 446 301 455 310 447 0 F 314 341 442 327 408 317 399 329 435 316 452 345 438 329 442 350 439 317 443 317 447 
B-1 

0 F 309 354 447 340 427 332 427 333 444 339 460 355 470 321 448 354 442 342 441 321 467 0 F 186 325 441 296 441 312 438 292 437 308 455 279 460 339 455 283 449 305 449 289 457 0 F 197 375 460 354 432 329 424 351 441 321 641 322 467 310 457 323 450 317 451 330 439 0 F 178 339 443 319 427 318 409 322 430 322 453 317 427 339 438 320 451 321 438 349 457 0 F 181 339 443 310 439 324 441 313 448 319 440 329 431 342 440 327 448 323 444 319 431 0 F 175 347 441 329 435 325 414 320 450 320 438 327 439 306 441 330 432 322 445 313 449 0 F 190 335 437 299 433 317 438 304 425 322 439 320 432 316 452 318 441 319 446 304 432 0 F 323 330 442 301 427 304 416 300 437 309 444 311 453 321 455 312 443 307 438 305 427 0 F 305 339 447 323 416 313 413 320 425 310 452 350 428 312 449 348 432 308 445 328 443 

30 M 94 323 430 314 433 325 448 324 442 30 M 93 311 432 321 430 326 464 327 455 30 M 90 321 437 345 440 333 431 329 432 30 M 80 342 440 350 443 314 463 317 441 30 M 78 317 443 331 435 350 412 348 440 30 M 76 331 430 311 445 299 455 303 442 30 M 407 299 433 323 440 307 435 302 448 30 M 402 308 441 320 432 325 432 321 452 30 M 417 327 435 330 434 328 444 322 438 30 M 406 312 447 317 432 340 430 338 437 30 M 92 318 436 313 440 319 458 322 430 30 M 88 325 427 325 432 337 464 339 429 30 M 95 300 449 310 445 325 454 332 432 30 M 91 336 422 300 420 337 448 341 437 30 M 77 315 436 328 436 312 426 319 442 30 M 83 333 427 319 434 324 432 329 440 30 M 404 308 429 340 430 309 448 301 439 30 M 100 326 435 325 428 343 446 342 448 30 M 99 358 445 315 434 341 440 335 453 30 M 97 349 434 320 432 331 415 335 453 30 F 81 315 444 320 430 311 451 323 342 310 457 302 454 332 448 329 402 313 441 314 420 30 F 96 340 439 365 431 350 452 335 340 348 458 361 451 330 440 299 455 341 439 353 442 30 F 89 332 457 336 428 333 407 350 345 330 440 341 435 315 452 337 429 332 452 323 445 30 F 85 348 445 334 429 334 433 342 343 333 452 336 445 319 441 332 443 331 461 324 434 30 F 403 333 449 303 440 316 433 356 439 317 441 315 453 340 457 321 438 322 459 310 436 
B-1 

30 F 405 363 443 337 425 338 445 342 352 342 463 344 438 322 442 339 448 339 456 327 451 30 F 408 321 451 309 424 309 435 336 344 315 454 313 410 330 455 319 451 319 445 305 437 30 F 427 352 449 334 426 342 443 306 3400 338 451 346 447 336 442 338 449 341 454 323 446 30 F 409 350 440 322 427 330 430 309 338 334 441 328 427 332 463 329 443 331 443 315 458 30 F 432 348 456 337 456 351 456 332 316 350 408 354 458 348 452 342 449 352 450 324 427 30 F 84 358 456 359 420 349 321 328 328 351 442 354 443 350 461 355 455 357 453 351 437 30 F 86 367 443 339 420 348 319 321 330 352 465 342 446 351 455 340 427 351 463 335 419 30 F 82 362 438 337 445 363 417 335 334 347 450 344 448 349 458 341 450 344 452 340 435 30 F 87 335 441 304 449 326 426 342 342 362 452 310 420 361 443 316 444 359 451 299 435 30 F 401 353 443 308 451 337 436 322 330 321 461 337 448 318 459 332 443 323 434 346 439 30 F 74 358 449 334 434 341 416 336 325 342 443 331 423 336 449 336 448 345 442 334 454 30 F 73 358 437 329 440 352 404 334 303 349 450 351 423 341 444 355 446 352 456 325 418 30 F 98 342 447 317 427 312 446 302 335 317 443 315 461 329 442 317 425 321 443 306 449 30 F 75 366 454 337 426 339 447 337 312 340 457 358 451 338 356 352 427 343 442 336 452 30 F 79 357 450 346 421 342 424 310 329 342 456 341 431 340 450 338 451 339 450 347 440 60 M 90 313 448 325 432 309 439 311 457 60 M 88 343 445 323 433 342 456 344 455 60 M 205 319 441 340 445 321 454 317 438 60 M 208 322 438 353 442 277 447 282 440 60 M 81 333 443 334 432 322 427 329 402 60 M 84 294 440 295 430 333 429 331 438 60 M 80 323 444 320 436 318 458 315 455 60 M 86 283 435 318 434 283 446 291 456 60 M 96 308 453 326 431 302 438 299 455 60 M 79 314 448 314 431 323 431 319 454 60 M 203 295 432 307 438 278 434 277 448 60 M 95 326 441 283 440 354 463 354 430 60 M 99 313 442 335 442 322 461 326 429 60 M 98 318 458 353 436 320 424 321 457 60 M 201 320 448 342 442 324 456 319 461 60 M 204 335 429 343 440 333 441 330 439 60 M 209 342 423 342 438 335 429 329 441 60 M 210 306 426 302 435 309 460 311 447 60 M 219 353 423 348 432 339 404 342 455 
B-1 

60 M 218 296 427 301 448 297 433 299 459 60 F 75 334 447 318 414 305 455 425 428 302 450 305 448 322 462 300 448 300 456 305 451 60 F 91 347 435 330 412 322 423 427 434 321 452 329 457 320 443 328 455 317 455 328 434 60 F 89 342 452 325 414 323 434 432 440 325 448 316 461 323 442 319 462 320 450 319 451 60 F 73 323 444 296 427 302 438 430 445 306 440 298 442 310 443 298 449 309 459 297 441 60 F 85 356 453 346 443 356 450 327 445 352 447 346 429 345 444 342 451 355 443 344 436 60 F 100 343 439 318 418 321 384 435 442 317 446 322 441 320 445 318 444 322 461 344 429 60 F 94 339 451 317 420 311 415 438 442 312 445 317 435 315 443 318 460 309 459 319 445 60 F 76 325 435 300 436 331 419 440 446 327 466 312 463 325 436 332 457 322 443 314 433 60 F 78 347 437 323 415 319 407 437 436 322 448 338 443 320 435 347 442 321 440 332 434 60 F 93 371 439 339 415 339 402 437 439 333 456 353 428 328 439 352 438 339 445 348 433 60 F 82 385 448 353 419 355 412 433 444 342 447 343 440 336 438 337 443 338 443 385 432 60 F 74 350 447 329 403 322 398 421 441 324 452 337 435 326 440 339 444 342 441 332 407 60 F 92 364 440 332 416 322 426 424 448 322 453 344 442 325 443 360 441 324 441 335 424 60 F 87 374 443 336 418 322 419 432 450 324 462 369 410 319 446 332 436 321 443 334 437 60 F 202 353 442 327 424 321 438 430 451 319 446 335 464 321 455 351 435 323 445 325 455 60 F 77 363 452 366 427 342 408 446 450 341 446 354 418 337 457 352 459 324 438 333 417 60 F 222 340 435 308 455 318 421 437 440 322 452 312 470 324 456 306 437 351 448 310 417 60 F 97 334 425 308 436 353 437 429 432 355 451 330 435 348 459 329 442 342 455 344 410 60 F 215 360 452 340 428 337 402 434 437 333 448 334 434 329 448 331 445 337 456 331 457 60 F 216 338 449 327 434 317 410 433 435 320 445 325 441 323 443 329 442 317 450 333 423 

125 M 75 321 437 320 430 327 440 331 447 125 M 77 281 423 284 433 313 439 319 442 125 M 95 328 408 325 447 317 445 313 448 125 M 233 323 448 327 444 326 465 320 441 125 M 100 335 425 330 425 323 474 326 452 125 M 81 333 419 332 422 319 457 323 449 125 M 98 315 444 317 425 290 448 320 451 125 M 97 300 447 299 435 337 464 332 439 125 M 208 328 430 325 430 303 455 310 443 125 M 76 296 433 291 432 308 466 316 441 125 M 235 324 416 319 442 321 436 318 444 125 M 93 324 422 324 435 315 420 316 447 125 M 82 321 420 319 440 305 431 309 442 
B-1 

125 M 94 333 432 332 422 308 434 307 450 125 M 90 321 410 327 415 292 456 317 442 125 M 99 317 414 320 412 320 429 315 443 125 M 78 348 423 341 431 343 445 342 436 125 M 85 308 414 310 422 301 444 309 439 125 M 204 318 438 315 421 312 444 308 440 125 M 79 338 420 334 417 331 441 325 435 125 F 88 336 430 303 443 336 416 308 440 358 438 339 456 355 434 335 453 332 468 310 449 125 F 87 347 431 326 414 324 448 320 441 334 443 348 433 339 440 339 448 329 450 345 442 125 F 89 376 435 352 427 347 411 349 442 342 455 335 452 337 439 342 443 341 457 316 416 125 F 84 348 440 316 446 341 399 318 432 328 456 337 431 331 445 346 438 332 458 323 414 125 F 207 338 440 309 421 312 413 312 430 342 449 309 459 345 449 312 439 339 451 317 462 125 F 202 349 439 315 440 324 439 316 436 337 462 329 456 337 459 317 436 338 452 320 460 125 F 259 376 438 350 401 340 414 345 447 315 443 356 451 319 456 349 440 317 448 343 440 125 F 209 331 441 316 445 342 413 317 422 321 440 316 415 320 451 315 452 322 441 327 429 125 F 258 367 452 361 429 362 426 357 421 345 443 360 445 340 464 358 451 342 442 351 428 125 F 260 340 449 319 442 334 432 320 433 341 445 363 427 354 462 350 446 341 441 312 456 125 F 74 388 445 363 434 359 422 357 428 352 443 369 420 351 442 352 437 351 452 336 439 125 F 80 346 438 328 414 328 417 333 439 325 436 345 425 332 454 350 447 323 443 317 436 125 F 83 340 437 308 407 309 413 310 442 310 445 349 423 312 445 332 452 312 446 325 436 125 F 73 348 433 316 396 314 414 317 438 311 438 336 442 310 455 348 448 310 450 325 436 125 F 206 374 441 331 423 335 418 331 445 336 440 335 424 329 456 341 455 334 457 329 430 125 F 205 357 433 317 421 323 421 322 425 327 461 326 423 334 447 330 447 332 456 337 430 125 F 234 369 437 346 423 343 423 338 423 339 448 345 466 337 444 342 442 341 457 359 430 125 F 201 388 436 377 433 357 426 376 446 362 4555 353 441 361 443 349 440 359 458 441 125 F 239 348 432 320 430 342 410 329 447 341 456 327 452 359 448 336 443 337 452 342 438 125 F 327 353 431 320 415 323 497 317 432 320 450 340 434 357 452 332 451 318 441 318 439 250 M 84 302 456 303 439 317 460 321 455 250 M 83 312 419 315 442 346 453 342 451 250 M 87 316 448 320 448 307 442 311 449 250 M 91 308 438 294 438 341 439 338 451 250 M 77 293 426 301 450 311 439 315 446 250 M 76 337 415 330 448 328 441 314 447 250 M 98 349 438 352 435 348 436 334 455 
B-1 

250 M 100 325 458 326 432 356 444 342 453 250 M 95 328 423 329 438 329 458 351 437 250 M 99 314 415 313 440 324 434 337 452 250 M 231 328 418 329 442 320 429 315 445 250 M 189 326 421 324 437 332 443 319 443 250 M 230 337 422 340 428 329 467 322 449 250 M 90 342 428 341 423 332 433 337 451 250 M 75 324 423 320 423 315 443 316 450 250 M 81 300 435 301 436 300 443 299 442 250 M 85 292 421 303 420 257 448 267 446 250 M 80 333 438 340 428 321 454 322 444 250 M 181 316 428 319 437 337 463 333 450 250 M 190 327 438 325 452 343 431 339 445 250 F 89 302 445 292 431 301 414 288 436 305 456 302 455 312 456 305 439 307 455 302 448 250 F 82 349 424 313 432 319 429 310 425 320 442 330 463 313 454 329 433 319 451 339 417 250 F 79 352 437 334 429 340 405 337 440 337 434 343 427 335 450 336 438 335 450 348 396 250 F 92 383 451 396 456 415 433 387 425 410 463 381 456 392 461 377 442 411 443 354 427 250 F 94 337 423 300 412 302 395 301 430 305 451 306 448 316 449 309 450 309 442 321 423 250 F 228 352 423 308 415 318 393 302 431 315 451 368 429 317 455 352 438 311 452 336 403 250 F 188 371 450 345 437 358 447 335 429 352 453 335 451 349 454 335 442 349 461 328 421 250 F 184 363 434 329 434 337 436 332 427 340 450 345 438 344 461 339 440 342 447 329 439 250 F 97 349 442 328 423 325 406 329 425 327 443 338 445 329 447 333 449 329 459 322 453 250 F 74 337 438 313 440 330 420 317 436 332 454 331 440 333 445 332 455 330 445 327 429 250 F 78 336 445 317 426 323 408 320 433 327 445 316 464 325 453 308 452 331 443 285 442 250 F 88 341 436 320 426 322 426 322 424 323 456 318 447 321 452 321 439 327 444 306 462 250 F 96 321 428 290 415 304 427 301 422 306 459 275 431 310 449 279 437 325 454 302 433 250 F 93 330 420 325 422 319 404 327 423 322 447 320 422 324 446 322 433 310 452 322 431 250 F 225 341 446 327 417 322 399 334 426 322 462 318 440 325 451 311 432 320 451 340 426 250 F 221 368 441 338 431 353 410 376 436 349 450 358 439 352 455 357 439 350 450 360 431 250 F 223 372 444 375 421 366 405 365 428 360 452 362 438 362 457 359 451 360 463 380 422 250 F 183 384 447 370 459 395 416 369 436 400 443 384 435 370 449 356 452 404 439 359 428 250 F 182 355 442 342 433 337 420 335 440 341 444 331 439 340 462 335 442 342 443 333 425 250 F 187 328 433 316 415 319 403 317 430 322 451 317 401 324 441 321 443 320 455 326 438 
B-1 

F2 egg production 

DOSE ID 12/21 12/22 12/23 12/24 12/25 12/26 12/27 

week 6 12/28 12/29 12/30 12/31 1/1 1/2 1/3 

week 7 1/4 1/5 1/6 1/7 1/8 1/9 1/10 

week 8 1/11 1/1

0 183 1 1 0 199 1 1 1 0 200 1 1 1 1 1 1 1 2 1 0 194 1 1 1 1 1 1 1 1 1 0 179 1 1 1 1 1 1 1 1 1 0 191 1 0 188 1 0 189 1 1 1 1 1 1 1 1 1 1 0 313 0 174 1 1 1 1 1 1 1 1 0 314 1 1 1 1 0 309 1 1 1 1 1 1 0 186 1 1 1 1 0 197 1 1 1 1 0 178 1 1 1 1 0 181 1 0 175 1 1 1 0 190 0 323 1 1 2 1 1 1 1 1 1 1 1 0 305 2 1 1 1 1 1 1 

30 81 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 96 1 1 1 1 1 1 1 1 1 1 30 89 1 1 1 1 30 85 1 1 1 1 1 1 30 403 30 405 1 1 1 1 1 1 30 408 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 427 1 1 1 1 1 1 1 30 409 30 432 30 84 1 1 1 1 1 1 1 1 1 30 86 1 1 1 1 1 
B-1 

30 82 1 1 30 87 1 1 1 1 1 1 1 1 30 401 30 74 1 1 1 30 73 1 30 98 1 1 30 75 1 1 1 1 1 1 1 1 1 1 1 1 1 30 79 1 1 1 1 1 1 1 1 1 1 1 60 75 1 1 1 1 1 1 1 1 60 91 1 1 1 1 1 1 1 60 89 60 73 1 1 60 85 1 1 60 100 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 94 1 1 1 1 1 1 1 1 1 1 1 60 76 60 78 1 1 1 1 60 93 1 1 1 1 1 1 1 1 1 60 82 1 1 1 1 1 1 60 74 1 1 1 60 92 1 1 1 1 60 87 1 1 1 60 202 60 77 60 222 60 97 2 2 1 1 2 1 1 1 1 1 1 1 1 60 215 1 1 1 1 1 1 60 216 1 1 1 1 

125 88 125 87 1 1 2 1 1 125 89 1 1 1 1 125 84 1 1 1 2 1 125 207 1 1 1 1 1 1 1 1 125 202 2 1 1 1 
B-1 

125 259 1 1 125 209 1 1 1 1 1 1 1 1 125 258 125 260 1 1 1 1 1 1 125 74 125 80 1 1 1 125 83 1 1 125 73 1 1 1 1 1 1 1 125 206 1 1 1 1 1 1 1 1 1 125 205 125 234 125 201 1 1 1 1 1 1 1 1 2 1 1 1 125 239 125 237 1 1 1 250 89 1 1 1 1 1 1 1 1 1 1 1 250 82 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 79 1 1 1 1 250 92 250 94 250 228 1 1 1 1 1 250 188 250 184 1 1 1 1 1 1 1 1 1 250 97 1 1 250 74 1 1 1 1 1 1 1 1 250 78 1 1 1 1 1 1 1 1 250 88 1 1 1 1 1 1 1 1 1 250 96 1 250 93 1 1 1 1 2 1 1 1 1 1 1 1 250 225 250 221 1 250 223 250 183 250 182 250 187 
B-1 

F2 egg production 

DOSE ID 1/18 1/19 1/20 1/21 1/22 1/23 1/24 week 10 1/25 1/26 1/27 1/28 1/29 1/30 1/31 week 11 2/1 2/2 2/3 2/4 2/5 2/6 2/7 week 12 2/8 2/9 

0 183 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 199 1 1 1 1 1 1 1 1 1 1 0 200 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 194 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 0 179 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 191 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 188 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 189 1 1 1 1 1 1 1 1 1 1 1 1 1 0 313 1 1 1 1 1 1 1 1 1 1 1 0 174 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 314 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 309 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 186 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 197 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 178 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 181 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 175 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 190 1 1 1 1 1 1 1 1 1 1 1 1 0 323 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 305 1 1 1 1 1 1 1 1 1 1 1 1 1 

30 81 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 96 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 89 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 85 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 403 1 1 1 1 1 1 1 1 1 1 1 1 1 30 405 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 408 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 427 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 409 1 1 1 1 1 1 1 1 1 1 1 1 1 30 432 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 84 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 86 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 
B-1 

30 82 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 87 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 401 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 74 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 73 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 98 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 75 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 79 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 75 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 91 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 60 89 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 73 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 85 1 1 1 1 1 1 1 1 1 1 1 60 100 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 60 94 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 76 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 78 1 1 1 1 1 1 1 1 60 93 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 82 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 74 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 92 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 87 1 2 1 1 1 1 1 1 1 1 1 60 202 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 77 1 1 2 2 1 1 1 1 1 1 1 1 1 1 1 60 222 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 97 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 215 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 216 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

125 88 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 125 87 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 125 89 1 1 1 1 1 1 1 1 1 1 1 1 1 125 84 1 1 1 1 1 1 1 1 1 1 1 1 2 125 207 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 202 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 
B-1 

125 259 1 1 1 1 1 1 1 1 1 1 1 1 1 125 209 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 258 1 1 1 1 1 1 1 1 1 1 1 1 125 260 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 74 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 80 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 83 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 73 1 1 1 1 1 1 1 1 1 1 1 1 125 206 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 205 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 234 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 201 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 125 239 1 1 1 1 1 1 1 1 1 1 1 1 125 237 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 89 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 82 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 79 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 92 1 1 250 94 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 228 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 188 1 1 250 184 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 97 1 1 1 1 1 1 1 1 1 1 1 1 1 250 74 1 1 1 1 1 1 1 1 1 1 1 1 250 78 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 88 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 96 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 93 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 225 1 1 1 1 1 1 1 1 1 1 1 1 1 250 221 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 223 250 183 250 182 250 187 1 1 1 
B-1 

F2 egg production 

DOSE ID 2/15 2/16 2/17 2/18 2/19 2/20 2/21 week 14 2/22 2/23 2/24 2/25 2/26 2/27 2/28 week 15 3/1 3/2 3/3 3/4 3/5 3/6 3/7 week 16 3/8 3/9 

0 183 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 199 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 200 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 194 1 1 1 1 1 1 1 1 2 2 1 0 179 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 191 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 188 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 189 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 313 1 1 1 1 1 1 1 1 1 1 1 1 1 0 174 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 314 1 1 1 1 1 1 1 1 1 0 309 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 186 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 197 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 0 178 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 181 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 175 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 190 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 323 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 305 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

30 81 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 96 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 89 1 1 1 1 1 1 1 1 1 1 1 1 1 30 85 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 403 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 405 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 408 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 427 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 409 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 432 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 84 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 86 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 
B-1 

30 82 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 87 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 401 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 74 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 73 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 30 98 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 75 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 79 1 1 1 1 1 1 1 1 1 1 1 1 1 60 75 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 91 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 89 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 73 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 85 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 100 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 94 1 1 1 1 2 1 60 76 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 78 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 93 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 82 1 1 1 1 1 1 1 1 1 1 1 1 1 60 74 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 92 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 87 1 1 1 1 1 1 1 1 1 1 1 1 1 60 202 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 77 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 222 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 97 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 215 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 60 216 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

125 88 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 87 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 89 1 1 1 1 125 84 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 207 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 202 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 
B-1 

125 259 1 1 1 1 1 1 1 1 1 125 209 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 258 1 1 1 1 1 1 1 1 125 260 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 74 1 1 1 1 1 1 1 125 80 1 1 1 1 1 1 1 1 125 83 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 73 1 1 1 1 1 1 1 1 125 206 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 205 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 234 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 201 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 239 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 237 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 250 89 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 82 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 79 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 92 1 1 1 1 250 94 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 228 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 188 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 184 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 97 1 1 1 1 1 1 1 1 1 1 1 250 74 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 78 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 88 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 96 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 93 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 225 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 250 221 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 250 223 250 183 250 182 250 187 1 1 1 1 1 1 1 1 1 1 1 1 
B-1 

F2 egg production 

DOSE ID 3/15 3/16 3/17 3/18 3/19 3/20 3/21 

week 18 3/22 3/22 3/23 3/24 3/25 3/26 3/27 3/28 3/29 3/30 3/31 4/1 

week 19 

0 183 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 199 1 1 1 1 1 1 1 1 1 1 1 0 200 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 194 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 179 1 1 1 1 1 1 1 1 1 1 1 0 191 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 188 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 189 1 1 1 1 1 1 1 1 1 0 313 1 1 0 174 1 1 1 1 1 1 1 1 1 1 1 1 0 314 1 1 1 1 1 1 1 1 1 1 1 1 1 0 309 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 0 186 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 197 1 1 2 1 1 1 1 1 1 1 0 178 1 1 1 1 1 1 1 1 0 181 1 1 1 1 1 1 1 1 1 1 1 1 0 175 1 1 1 1 1 1 1 1 1 0 190 1 1 1 1 1 1 1 1 1 1 1 1 1 0 323 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 305 1 1 1 1 1 1 1 1 1 1 1 

30 81 1 1 1 1 1 2 1 1 30 96 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 89 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 85 1 1 1 1 1 1 1 1 1 1 1 1 1 30 403 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 405 1 1 1 1 1 1 1 1 1 1 1 30 408 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 427 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 409 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 432 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 84 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 86 1 1 1 1 1 1 1 1 1 1 1 1 1 
B-1 

30 82 1 1 1 1 1 1 1 1 1 1 1 30 87 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 401 1 1 1 1 30 74 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 73 1 1 1 1 1 1 1 1 1 1 30 98 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 30 75 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 30 79 1 1 1 1 1 1 1 1 1 1 1 60 75 1 1 1 1 1 1 1 1 1 1 1 1 60 91 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 89 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 60 73 1 1 1 1 1 1 1 1 1 1 60 85 1 1 1 1 1 1 1 1 1 1 60 100 1 1 1 1 1 1 1 1 1 1 60 94 1 1 1 60 76 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 78 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 93 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 60 82 1 1 1 1 1 1 1 1 1 60 74 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 92 1 1 1 1 1 1 1 1 1 1 1 1 60 87 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 202 1 1 1 1 1 2 1 1 1 1 1 1 60 77 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 222 1 1 1 1 1 1 1 1 1 1 1 1 1 60 97 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 215 1 1 1 1 1 1 1 1 1 1 1 1 1 1 60 216 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

125 88 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 87 1 1 1 1 1 1 1 1 1 125 89 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 84 1 1 1 1 1 1 1 1 1 1 1 125 207 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 125 202 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 
B-1 

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B-1