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Intra-Laboratory Analysis Report for Mysid: UNT Vinclozolin Data 

Endocrine Disruptor Screening Program 

Contract No: EP-W-11-063 

Task Order No: 2 

Statistical Support for Technical Directive #3 

Data Analysis Support for Invertebrate Tier 2 Test and Generation of 

Integrated Summary Report 

Intra-Laboratory Statistical Analysis for UNT Vinclozolin in Mysids 

U. S. Environmental Protection Agency 

May 17, 2013 

BATTELLE 

505 King Avenue 

Columbus, Ohio 43201 
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Table of Contents 

Page

1. Introduction ................................................................................................................ 1 

2. Study data ................................................................................................................... 2 

2.1 Data Endpoints ........................................................................................................... 2 

2.2 Data Issues and Data Handling .................................................................................. 3 

2.3 Analysis Considerations ............................................................................................. 6 

3. Statistical Methods ..................................................................................................... 8 

3.1 Comparisons of test chemical with the control .......................................................... 8 

3.1.1 Outlier Screens ........................................................................................................... 8 

3.1.2 Normality ................................................................................................................... 9 

3.1.3 Transformation of data ............................................................................................... 9 

3.1.4 Heterogeneous variances ............................................................................................ 9 

3.1.5 Monotonicity of response ........................................................................................... 9 

3.1.6 Comparisons of test chemical to controls ................................................................... 9 

3.2 Comparisons of the F0 and F1 generations .............................................................. 11 

3.2.1 Outlier screens .......................................................................................................... 11 

3.2.2 Normality ................................................................................................................. 12 

3.2.3 Transformation of data ............................................................................................. 12 

3.2.4 Heterogeneous variances .......................................................................................... 12 

3.2.5 Monotonicity Test .................................................................................................... 12

3.2.6 F0 generation versus F1 generation .......................................................................... 12 

4. results ....................................................................................................................... 13 

Tables

Table 0. Data information for UNT Vinclozolin ...................................................................... 4 

Table 1A. Summary of Reproduction Endpoints. Laboratory: UNT, Test Chemical: 

Vinclozolin, Generation: F01,2,3 ................................................................................ 14 

Table 1B. Summary of Reproduction Endpoints. Laboratory: UNT, Test Chemical: 

Vinclozolin, Generation: F11,2,3 ................................................................................ 15 

Table 1C. Statistical Analysis Results for Reproduction Endpoints. Laboratory: UNT, Test 

Chemical: Vinclozolin, Generation: F0 .................................................................... 16 
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Table 1D. Statistical Analysis Results for Reproduction Endpoints. Laboratory: UNT, Test 

Chemical: Vinclozolin, Generation: F1 .................................................................... 17 

Table 1E. Comparisons between Two Generations for Reproduction Endpoints. Laboratory: 

UNT, Test Chemical: Vinclozolin. .......................................................................... 18 

Table 2A. Summary of Growth Endpoints. Laboratory: UNT, Test Chemical: Vinclozolin, 

Generation: F01,2,3,4,5 ................................................................................................. 19 

Table 2B. Summary of Growth Endpoints. Laboratory: UNT, Test Chemical: Vinclozolin, 

Generation: F11,2,3,4 ................................................................................................... 20 

Table 2C. Statistical Analysis Results for Reproduction Endpoints. Laboratory: UNT, Test 

Chemical: Vinclozolin, Generation: F0 .................................................................... 21 

Table 2D. Statistical Analysis Results for Reproduction Endpoints. Laboratory: UNT, Test 

Chemical: Vinclozolin, Generation: F1 .................................................................... 22 

Table 2E. Comparisons between Two Generations for Growth Endpoints. Laboratory: UNT, 

Test Chemical: Vinclozolin. ..................................................................................... 23 

Table 3A. Summary of Survival Endpoints. Laboratory: UNT, Test Chemical: Vinclozolin, 

Generation: F01,2,3 ..................................................................................................... 24 

Table 3B. Summary of Survival Endpoints. Laboratory: UNT, Test Chemical: Vinclozolin, 

Generation: F11,2,3 ..................................................................................................... 25 

Table 3C. Statistical Analysis Results for Reproduction Endpoints. Laboratory: UNT, Test 

Chemical: Vinclozolin, Generation: F0 .................................................................... 26 

Table 3D. Statistical Analysis Results for Reproduction Endpoints. Laboratory: UNT, Test 

Chemical: Vinclozolin, Generation: F1 .................................................................... 27 

Table 3E. Comparisons between Two Generation Survival Endpoints. Laboratory: UNT, Test 

Chemical: Vinclozolin. ............................................................................................. 28 

Figures

Figure 1. Time to Mature By Generation and Group For Chemical Vinclozolin 

From Lab UNT. ........................................................................................................ 29 

Figure 2. Number Offspring (brood 1) By Generation and Group For Chemical 

Vinclozolin From Lab UNT. .................................................................................... 29 

Figure 3. Number Offspring (brood 2) By Generation and Group For Chemical 

Vinclozolin From Lab UNT. .................................................................................... 30 

Figure 4. Number Offspring (two broods) By Generation and Group For Chemical 

Vinclozolin From Lab UNT. .................................................................................... 30 

Figure 5. Days to Brood 1 By Generation and Group For Chemical Vinclozolin 

From Lab UNT. ........................................................................................................ 31 
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Figure 6. Days to Brood 2 By Generation and Group For Chemical Vinclozolin 

From Lab UNT. ........................................................................................................ 31 

Figure 7. Days Between Broods 1 and 2 By Generation and Group For Chemical 

Vinclozolin From Lab UNT. .................................................................................... 32 

Figure 8. Percent of Females Productively Active By Generation and Group For 

Chemical Vinclozolin From Lab UNT. .................................................................... 32 

Figure 9. Sex Ratio By Generation and Group For Chemical Vinclozolin From 

Lab UNT. ................................................................................................................. 33 

Figure 10. Dry Weight (mg) (day 0) By Generation and Group For Chemical 

Vinclozolin From Lab UNT. .................................................................................... 33 

Figure 11. Dry Weight (mg) (day 7) By Generation and Group For Chemical 

Vinclozolin From Lab UNT. .................................................................................... 34 

Figure 12. Dry Weight (mg) (day 14) By Generation and Group For Chemical 

Vinclozolin From Lab UNT. .................................................................................... 34 

Figure 13. Dry Weight (mg) (Termination) By Generation and Group For Chemical 

Vinclozolin From Lab UNT. .................................................................................... 35 

Figure 14. Length (mm) (day 0) By Generation and Group For Chemical Vinclozolin 

From Lab UNT. ........................................................................................................ 35 

Figure 15. Length (mm) (day 7) By Generation and Group For Chemical Vinclozolin 

From Lab UNT. ........................................................................................................ 36 

Figure 16. Length (mm) (day 14) By Generation and Group For Chemical Vinclozolin 

From Lab UNT. ........................................................................................................ 36 

Figure 17. Length (mm) (Termination) By Generation and Group For Chemical 

Vinclozolin From Lab UNT. .................................................................................... 37 

Figure 18. Reproduction Survival Rate (%) (day 7) By Generation and Group For 

Chemical Vinclozolin From Lab UNT. .................................................................... 37 

Figure 19. Reproduction Survival Rate (%) (day 14) By Generation and Group For 

Chemical Vinclozolin From Lab UNT. .................................................................... 38 

Figure 20. Reproduction Survival Rate (%) (Termination) By Generation and Group 

For Chemical Vinclozolin From Lab UNT. ............................................................. 38 

Figure 21. Total Survival Rate (%) (day 7) By Generation and Group For Chemical 

Vinclozolin From Lab UNT. .................................................................................... 39 

Figure 22. Total Survival Rate (%) (day 14) By Generation and Group For Chemical 

Vinclozolin From Lab UNT. .................................................................................... 39 

 
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1. INTRODUCTION 

 To evaluate the effects of endocrine disrupting chemicals, EPA is currently developing and 

validating a Tier 2, two-generation mysid toxicity test to characterize the adverse consequences of 

exposure to putative endocrine active substances. The test is being carried out in three labs: SMV, UNT, 

and WLI with various chemicals. UNT conducted a two-generation mysid survival and growth test with 

vinclozolin. The draft analysis documented in this report is based on this UNT vinclozolin test. 

 The two-generation test is initiated with newly-released (<24 hours old) offspring (F0 generation) 

that are grown to sexually maturity (~13 to 16 days) and then their offspring are collected (F0 offspring). 

The F1 generation test is started with newly hatched F0 offspring. The F1 generation is also grown to 

maturity and their offspring are collected (F1 offspring). The test is conducted with five graded 

concentrations of vinclozolin and an untreated control, with three replicate tanks per test concentration or 

control group. Each replicate tank contains two retention baskets, one basket housing mysids for 

reproduction observations and a second basket for growth and optional biochemical measurements. 

 For the F0 test the growth and reproduction baskets each initially include 20 newly hatched 

mysids. With the appearance of a marsupial pouch on females (sexual maturity), mature females are 

isolated as pairs with males in separate brood cups. Seven breeding pairs are selected in each replicate 

reproduction basket. The exposure of these F0 mysids is continued until two weeks after the median 

time to the second brood release in the control group. Offspring (F1 generation) are collected from the 

F0 generation each morning from the brood cups and transferred immediately to baskets with the same F0 

exposure conditions as their F0 parents. 

 The F1 generation is begun with newly hatched F0 offspring. Within each replicate tank the 

growth and reproduction baskets each initially include 15 newly hatched mysids. With the appearance of 

a marsupial pouch on females (sexual maturity), mature females are isolated as pairs with males in 

separate brood cups. Seven breeding pairs are selected in each replicate reproduction basket. The 

exposure of these F1 mysids is continued until two weeks after the median time to the second brood 

release in the control group. Offspring are collected from the F1 generation each morning from the brood 

cups and transferred immediately to baskets with the same exposure conditions as their F1 parents. 

 Development, sexual maturation, reproduction, growth, and survival endpoints are observed in 

the F0 and F1 generations. Measurements are made of a number of endpoints in both F0 and F1 

generations reflective of functional endocrine regulation of development, growth, reproduction, and 

survival. They include sex ratio, time to maturity, time to brood release for the first two broods, and 

interbrood duration between the first two broods. Observations are assessed and recorded daily for each 

individual or breeding pair on mortality, sex determination, reproductive condition of female, and number 

of young released in each of the first two broods. Mysids are sampled to determine weight and length at 

the beginning (Day 0) before being placed into the growth and reproduction baskets, and then at weekly 

intervals (Day 7 and Day 14) from the growth baskets. At termination (around Day 32 for UNT 

vinclozolin), mysids from the reproduction baskets are measured for weights and lengths. 

Intra-laboratory statistical analysis of the UNT vinclozolin mysid study was performed according 

to a common statistical analysis plan. For this intra-laboratory analysis comparisons were made between 
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the test chemical concentration groups and the concurrent control group to assess the mysid responses to 

graded concentrations of vinclozolin. The results of the two-generations were compared to assess the 

benefits of a two-generation study as compared to a one-generation life cycle study. 

2. STUDY DATA 

2.1 Data Endpoints 

The test method specified 25 endpoints. Some of the endpoints were recorded per mysid or 

composite of mysids (e.g. day 0 weight), some were recorded per breeding pair, and some were recorded 

per basket (i.e. per replicate tank). These endpoints were divided among three categories: 

(i) Reproduction (F0, F1) (11)

a. Sex ratio (per basket) 

b. Time to mature (per basket) 

c. Time to first brood release (per breeding pair) 

d. Time to second brood release (per breeding pair) 

e. Interbrood duration, time between release of first and second brood (per breeding pair) 

f. Number of young per female in the first brood (per breeding pair) 

g. Number of young per female in the second brood (per breeding pair) 

h. Total number of young in the first two broods per female (per breeding pair) 

i. Percentage of females that are reproductively active in the first two broods (per basket) 

j. Total reproduction days in the first two broods (per breeding pair) 

k. Number of young per female per reproductive day in the first two brood (per breeding 

pair) 

 Total reproduction days in the first two broods (per breeding pair) and number of young per 

female per reproductive day in the first two brood (per breeding pair) were not provided for the UNT 

vinclozolin study Excel spreadsheets. They could be calculated based on the reproduction data, but that 

has not been done yet. 

(ii) Survival (F0, F1) (6) 

a. Survival rate for reproduction animals at days 7, 14 and termination (reproduction 

survival, per basket)1

b. Survival rate for all animals including both the growth and reproduction animals at days 7 

and 14 (total survival, per tank) 

 For the growth basket mysids the number removed for weight and length determinations were 

adjusted for (i.e. were omitted from both the numerator and denominator) in the survival rate calculations. 

Reproductive survival was determined among the animals assigned to the brood chambers. Total survival 

at termination was not determined due to the absence of appropriate data. 

 

1 The reproduction termination survival rates were based only on the 14 adult reproduction mysids in the seven brood cups 

starting on day 14, as shown in the survival and reproduction Excel data. Mortality before day 14 was not accounted for in the

survival rates. 
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(iii) Growth (F0, F1) (8)2, 3 

a. Length at day 0 (per mysid) 

b. Length at day 7 (per mysid) 

c. Length at day 14 (per mysid) 

d. Length at termination. (per mysid) 

e. Weight at day 0 (on average per composite of 5 mysids) 

f. Weight at day 7 (on average per composite of 2 mysids) 

g. Weight at day 14 (per mysid) 

h. Weight at termination (per mysid). 

 Note that on day 0, the newly hatched F0 generation mysids were kept in untreated water, By 

contrast on day 0 the newly hatched F1 generation mysids were kept in water at the same test chemical 

concentration as their parents’ treatment groups. Length and weight measurements were determined for 

mysids on day 0 before they were assigned to growth and reproduction baskets, for mysids from the 

growth baskets at day 7 (6 per replicate tank) and day 14 (all surviving mysids on day 14), and for the 

surviving reproduction mysids at termination. For the weight data, there were seven composites for the F0 

generation (all in the control) and a total of 53 composites (9 composites per concentration groups (8 for 

the highest concentration group)) for the F1 generation on day 0. There were no replicate tank identity 

designations in the growth data on day 0. On day 7, there were 3 composite weights per replicate for each 

of the three replicates per concentration group. Length and weight measurements were not collected for 

F1 on day 7 in the Excel data sheets. 

 Each laboratory ran a concurrent control and five targeted test concentration levels. The actual 

exposure concentration levels in general differed from the targeted levels. The statistical analysis 

procedures do not utilize the actual concentration levels. The concentrations are labeled by their targeted 

levels but the labels are not actually used in the analyses. 

2.2 Data Issues and Data Handling 

Data were received in Excel table format not directly suitable for analysis. They were 

reformatted into STACHARRMS friendly formatted spreadsheets. SAS and STATA programs were used 

to read in the raw data, define program friendly variable names, and add new character variables to the 

data (laboratory name, chemical name, generation). The reformatted data sheets were saved into three 

files according to the endpoint category: reproduction, growth, and survival. 

 

 

2

 Day 14 and terminal weights and lengths were not reported by gender. 3

 Weight and length results were reported pooled across replicate tanks within control or test concentration groups 
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Table 0. Data information for UNT Vinclozolin 

Data Files and data sheets Endpoints Comments 

Original - Vinlozolin 

Spreadsheet UNT.xlsx: 

− Time to 1st Brood (per 

matching pair) 

− Time to 2nd Brood (per 

matching pair) 

− F0 and F1 Interbrood 

Duration (per matching pair 

− % Reproducing F0 and F1 

(per replicate tank) 

− % Reproducing F0 and F1 

 (per replicate tank) 

− % Females with 1st+2nd 

Brood 

 (per concentration, for each 

of the two broods) 

− Time to Maturation (per 

replicate tank) 

− Sex Ratio 

(per replicate tank) 

 Mysid Vinclozolin 

Reproduction.xlsx

# of offspring in each brood and 

total over two broods: one data 

sheet for a chemical 

concentration per generation. 

Reformatted – 

“UNTVinReproCharrms.xlsx” 

- One datasheet per endpoint 

per generation 

 Time to Maturity 

 Time to 1st and 2nd broods

 # of offspring at each 

brood 

 time between two broods 

 # of offspring at two 

broods 

 %of female 

reproductively active 

 Sex ratio 

The following two variables 

are not provided: 

 Offspring per female per 

reproductive day for the 

first two broods 

 Total reproductively 

active days for the first 

two broods 

 If a mysid in a matching pair died 

before the first or second brood, the 

0 brood sizes were recorded in the 

original data files. Per EPA 

suggestion, brood size values were 

changed as: zero if female survived, 

and missing for a brood (1st or 2nd 

brood) if female died without 

releasing the brood. 

 The total offspring for the first two 

broods was recorded as the sum of 

the two broods in the original data 

sheet treating the missing as zero. 

Thus it needed to be recalculated 

such that a missing value is 

assigned if there is a missing brood 

size in either brood 1 or brood 2. 

 All changes described above are 

recorded in the reformatted data 

files. 

 % of female reproductively active, 

and sex ratios are reported per tank. 

The other reproductive endpoints 

are reported per mating pair. 

 Replicate tank IDs are given 

(labeled as 1, 2, 3). 

 Sex ratios were originally presented 

as % of females over all animals in 

the original data file. They were 

redefined as #males/#females in the 

reformatted data. 

Vinlozolin Spreadsheet 

UNT.xlsx:

− F0 Day 0 Mysid Lgth and 

Wt.; 

− F0 Day 7 Mysid Lgth and 

Wt.; 

− F0 Day 14 Mysid Lgth and 

Wt.; 

− F0 to Term Mysid Lgth and 

Wt. 

− F1 Day 0 Mysid Lgth and 

Wt.; 

− F1 Day 14 Mysid Lgth and 

Weight (dry weight) and 

length 

Weight (dry weight) and 

length for F1 Day 7 are not 

collected 

 Lengths were measured per mysid 

from day 0 to termination. 

 Weights were measured per 

composite of 5 mysids on day 0 and 

per composite of 2 mysids day 7 

(on day 7, per pair on smv), per 

mysid on day 14 and termination. 

 Replicate (basket) ID are not 

available on day 0. Replicate 

(basket) ID are available (replicate 

ID given on day 7) or color-coded 

on day 14 and at termination). 

 There are no growth data for day 7 
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Data Files and data sheets Endpoints Comments 

Wt.; 

− F1 to Term Mysid Lgth and 

Wt. 

for F1 generation. 

 Sex information for day 14 and 

termination are given. Data are not 

analyze by sex. 

Original- Survival and 

Reproduction Vinclozolin.xlsx 

- F0 Survival and Repro. 

- F1 Survival and Repro. 

Reformatted- UNT Vinclozolin 

Survival

- F0Survival 

- F1Survival 

 Survival (reproduction) 

(day 7, day 14 and 

termination) 

 Survival (total) (day 7, 

day 14) 

 Survival (total) at 

termination is not 

available4

. 

 Reformatted survival data were 

hand entered using the information 

from the original files. The original 

files have survival data for both 

growth and reproduction mysids 

from day 0 to day 14. Thus the 

survival data can be determined 

using the information on days 7 and 

14. 

 The initial numbers of mysids are 

20 for F0 and 15 for F1 per tank. 

Thus there should not be more than 

20 mysids in a tank. if there are 

more than 20 mysids alive on day 7, 

we treated them as 20. 

 Sometimes more mysids survived at

a later date than at a previous date. 

For example, if on day 7, the 

number alive was recorded as 19 

but 20 were alive on day 14, we 

assume survival number on day 7 as 

20 instead of 19. 

 There are no survival records for 

mysids that were not assigned to the 

brood chambers after day 14. The 

reproduction mysid survival for 

termination is determined using the 

animals from the brood chambers 

only. This didn’t adjust for the 

mortality rate before days 14, when 

they were paired and assigned to the 

brood chamber as we could not 

determine the survival status of 

mysids that were not assigned to the 

brood chambers. This survival rate 

could be overestimated. 

 On the reproductive basket, there 

are fewer than 14 living mysids in 

some replicates on day14, however 

there are 14 or more than 14 mysids 

 

4

 The survival worksheet in the Vinclozolin Excel spreadsheet is problematic to use for termination survival. The numerator 

values are inconsistent with survival information in the Survival and Reproduction spreadsheet. The denominator values are not

adjusted for non-treatment related mortality, in particular for the growth mysids that are removed at day 7 and day 14 for 

determination of lengths and weights. 
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Data Files and data sheets Endpoints Comments 

(7 pairs of mysids in the brood 

chambers for the replicates). The 

data are used as they are. 

 If there is a male death, which is 

assumed if number of animals goes 

from 2 to 1 and then back to 2, one 

add-in animal should be included in 

the survival pool. If there is a 

female death, which is assumed if 

number of animals goes from 2 to 1 

and then stops (study for the brood 

chamber was terminated), one dead 

and one live are counted. 

 If survival/reproduction data for 

two animals in a brood pair ended 

at the same time before the terminal 

day, they are assumed to be dead 

 Survival endpoints are reported per 

replicate tank. 

2.3 Analysis Considerations 

Several analysis considerations need to be noted. These are discussed below. 

 STATCHARRMS uses totally independent statistical procedures for analyses within the F0 and 

F1 generations as compared to comparisons between the F0 and F1 generations. Monotonicity 

tests and concentration trend tests are only made available under the one-way ANOVA in 

STATCHARRMS. Thus for the analyses within generations it was elected to use the one-way 

ANOVA procedure. In the one-way ANOVA procedure, STATCHARRMS pools the results 

across the test or control groups. The one-way ANOVA (parametric or nonparametric forms) 

was applied to the one generation data and there are no random replicate effects. For the 

comparison between the F0 and F1 generations STATCHARRMS utilizes a two-way mixed 

effects analysis of variance, with replicate tanks within treatment groups treated as random effects. 

The objectives of the analyses differ. The analyses within generations compare the test group 

results to the control group results. The analyses between generations compare the results 

between generations, either pooled across groups or separately within test or control groups. 

 The preliminary tests for outliers, normality, heterogeneity of variance, and monotonicity were 

carried out independently for the analyses within the F0 generation, the analyses within the F1 

generation, and the analyses between the F0 and F1 generations. There is no test for 

monotonicity within the two-way AVOVA for comparisons between generations. The results of 

the preliminary tests may differ among the F0 analysis, the F1 analysis, and the analysis for 

comparison between F0 and F1. Thus the approaches for the principal tests used are 

independently determined among these analyses. For example, for a particular endpoint some of 
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the analyses may be parametric for one-generation and nonparametric for the other generation, or 

they may be based on different transformations of the endpoint (or none). 

 STATCHARRMS uses independent outlier detection procedures with the one-way ANOVA and 

with the two-way ANOVA. Therefore the potential outliers screened out may not necessarily be 

consistent among the F0 and F1 within-generation analysis (one-way ANOVA), and the F0 and 

F1 comparison between-generation analyses (two-way ANOVA). The statistical potential outliers 

are screened values based on the statistical model applied, and they may not be in fact 

biologically important. The potential outliers from both one-way and two-way ANOVA’s should 

be jointly considered. EPA will determine whether the flagged values should be included in the 

subsequent analyses or should be treated as outliers. 

 The statistical analysis plan calls for the screened values to be sent to EPA for review. EPA 

would determine which screened values to include in the analyses, which to exclude from the 

analyses, and which to both include in and exclude from the analyses. In the interests of time for 

the draft analyses all the screened values were included in the analyses. The screened values 

however were enumerated in the STATCHARRMS outputs and will be sent to EPA for review. 

 Survival for the reproduction animals at termination is calculated based on the animals assigned 

to and later added into the brood chambers, as showed in the Survival and Reproduction 

Vinclozolin Excel file. This accounts for lethality only within the seven or less breeding pairs 

from day 14 to termination day. It does not account for lethality among the reproductive mysid 

prior to being paired at day 14. 

 Total survival at termination is not determined. The Pre and Post Paring Survival worksheet in 

the Vinclozolin UNT Excel spreadsheet are not used in the analysis because they are inconsistent 

with lethality shown in the Survival and Reproduction Vinclozolin Excel spreadsheet. 

 Survival for both reproduction and growth animals on day 7 and day 14 are determined based on 

the data in the Survival and Reproduction Vinclozolin Excel file. 

 For the reproductive response when 0 is reported as a brood size the question arises as to whether 

or not to treat this as a missing value or as a 0 numerical value in the analysis. EPA has 

suggested that if the female is living the 0 brood sizes should be counted as 0 and included in the 

brood size averages. If the female died before the recorded 0 size is generated, the brood size 

should be counted as “NA” (or recorded as a missing). Battelle has changed the data entries in the 

columns for brood size 1 and brood size 2 in the Excel spreadsheet according to this suggestion. 

In the reformatted data, the changes are also made to the affected endpoints: total number of 

offspring in the first two broods, and the number of offspring per female per reproductive day 

during the two broods. If the data entries for a given brood size is 0 or “NA”, the times for the 

brood was also recorded as “NA” in the original data file and they are treated as missing in this 

analysis. If the brood size is missing in either brood 1 or brood 2, then the sum of the two broods 
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is also treated as missing. This applies to endpoints: number of offspring in brood 1, number of 

offspring in brood 2, number of offspring in broods 1 and 2, and percent reproductive females. 

 Total reproduction days in the first two broods (per breeding pair) and number of young per 

female per reproductive day in the first two brood (per breeding pair) are not available for the 

UNT vinclozolin study. 

3. STATISTICAL METHODS 

Statistical analysis procedures used conform to the statistical analysis procedures specified in the 

STATCHARRMS package. Statistical analyses include two components: comparisons of test chemical 

with the control within each generation and comparison of the F0 and F1 generations. The statistical 

analysis was carried out in a univariate fashion independently for each endpoint. 

Statistical summaries and statistical analyses were applied to the data as reported without first 

summarizing within baskets. For example if the data were reported on a per mysid basis then the analysis 

procedures were applied to per mysid data without first averaging across the mysids in the same replicate 

baskets. 

3.1 Comparisons of test chemical with the control 

 Mysid data for the multi-concentration test chemical were analyzed according to a common 

procedure for each generation. Analyses were carried out on a per mysid, per breeding pair, or per basket 

basis depending on how the data were reported. Preliminary tests were first carried out to detect outliers, 

departure from normality, heterogeneity of variance, and departure from monotonicity of response. 

Subsequent comparisons among groups were carried out including all the values (including the screened 

outlier values) using procedures depending on the outcomes of the preliminary analyses. All the test 

procedures described here are based on the default auto option programs in STATCHARRMS. 

3.1.1 Outlier Screens 

 For each generation, outlier screens in the one generation analyses were carried out using Tukey’s 

outlier rule on the untransformed data. Outliers were assessed from residuals (results of subtracting 

treatment group means from individual values) to avoid confounding outliers and treatment effects. Any 

response more than 1.5 times the interquartile range above the third quartile (75th percentile) or below the 

first quartile (25th percentile) is considered an outlier by Tukey’s rule. 

 The statistical analysis plan states that the results of the statistical outlier screens are to be 

documented and reported to EPA for review to determine if the deviations, although statistically screened, 

are in fact biologically unimportant. EPA will determine whether the flagged values should be included 

in the subsequent analyses or should be treated as outliers. The subsets of potential outliers are responsespecific. Due to time constraints, the statistical summaries and analyses in this draft report were carried 

out including the flagged values. The list of screened values is included in the STATACHARRMS 

outputs and will be sent to EPA. 
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3.1.2 Normality 

 Normality was formally assessed in the ANOVA procedure on the untransformed data using the 

Shapiro-Wilk test (Shapiro and Wilk, 1965). It was assessed from residuals (results of subtracting 

treatment group means from individual values). Screened potential outliers were included in the tests. 

3.1.3 Transformation of data 

 STATCHARRMS includes three data transformations: log-transformation, square root 

transformation, and arc-sin square-root transformation. Under the auto option in STATCHARRMS, the 

analysis procedure is first applied to the untransformed data. If the normality assumption is valid, then no 

transformation is carried out. If the normality assumption is invalid, then normality tests are again 

performed on the log and square-root transformed data. If the normality assumption is valid after either 

transformation, then further procedures are applied to the transformed data. If the normality assumption 

is invalid for both the untransformed data and the transformed data, then nonparametric procedures are 

applied. Transformation is not necessary for nonparametric procedures as any transformation does not 

change the rank scores for the data when nonparametric procedures are used. 

3.1.4 Heterogeneous variances 

If the normality assumption was accepted for either the untransformed or the transformed data 

heterogeneity of variances among the concentration groups was assessed in the ANOVA procedure on the 

untransformed or the transformed data with Levene’s test with absolute residuals (Levene, 1960). 

Screened outliers were included in the Levene’s tests. If the normality assumption was invalid for both 

the untransformed and the transformed data heterogeneity of variances among the concentration groups 

was not assessed, as nonparametric procedures were applied. 

3.1.5 Monotonicity of response 

 In STATCHARRMS, a monotonicity test using linear and quadratic contrasts under one-way 

ANOVA was applied to normalized rank statistics (to avoid the complications that can arise from nonnormal or heterogeneous data). If the linear contrast was significant but the quadratic contrasts was not 

significant, then that was taken as evidence of monotonicity.

3.1.6 Comparisons of test chemical to controls 

Depending on the results of the test for monotonicity either ordered alternatives or general 

alternatives analysis procedures were used. Ordered alternatives procedures assume that the response is 

monotonically increasing or monotonically decreasing with increasing test chemical concentration. 

Ordered alternatives procedures were always carried out on the ranks of the data. If there was significant 

departure from monotonicity, general alternatives procedures were carried out. 

If the test for departure from normality was not significant for either the untransformed or the 

transformed data, normal theory parametric methods with homogeneous or heterogeneous variances were 

used. If the test for departure from normality was significant for both the untransformed and the 

transformed data nonparametric methods were used. The analysis procedures fall into three categories: 
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 Ordered alternatives − nonparametric 

 General alternatives − parametric 

 General alternatives − nonparametric 

Ordered alternatives – step down procedure 

 If the hypothesis of monotonicity of response was accepted, ordered alternatives procedures were 

used. Step down multiple comparison procedures are particularly useful for ordered alternatives 

situations. The test is carried out as long as there are significant effects, starting from all concentration 

groups (including the control), successively omitting the highest concentration group. If at any stage the 

comparison is nonsignificant, the process stops and all means in that concentration group and lower are 

declared to be compatible with the null hypothesis of equality to the control group mean. No adjustment 

for simultaneity is required. In STATCHARRMS, the nonparametric Jonckheere-Terpstra (Jonckheere 

(1954)) test of trend was used on the rank data. 

 Additionally, global overall concentration effects were tested. These were based on the 

parametric analysis of variance F-test if the normality assumption held or the Kruskal-Wallis test if the 

normal assumption did not hold for both the untransformed and the transformed data. See the general 

alternative for more details. 

General alternatives – parametric procedure 

 If the hypothesis of monotonicity of response was rejected and the normality assumption was not 

rejected, parametric general alternative procedures were used. If the overall concentration effect with the 

ANOVA was significant, comparisons of test concentrations to the control were conducted using 

Dunnett’s test with homogeneous within-treatment variances and using the Tamhane-Dunnett test with 

heterogeneous within-treatment variances. 

General alternatives – Nonparametric procedure 

 If the hypothesis of monotonicity of response was rejected, and the normality assumption was 

rejected for both the untransformed and the transformed data, nonparametric general alternative 

procedures were used. The Kruskal-Wallis test was applied to test for the overall concentration effect. If 

the overall concentration effect was significant, the comparisons of test concentration to the control were 

conducted using a modified Dunn’s test. 
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Exact Permutation Tests 

 For nonparametric procedures, if a large percentage of the responses have the same measured 

value (ties), specifically in the case of very small samples, say less than three per concentration for most 

concentrations, it is may not be possible to assess in a meaningful way the true distribution of responses 

across test concentration groups. One can either use an assumed distribution or treat this situation in the 

same way as data with massive ties. Exact permutation test methodology is an appropriate tool to use in 

these cases. The ANOVA component of STATCHARRMS automatically uses exact methods when 

deemed appropriate. In the ANOVA procedure, there are two conditions under which exact permutation 

methods are used: (1) There are no more than three experimental units in any treatment group and nonparametric procedures are found appropriate. (2) There are “massive ties” in the responses, i.e., 25% or 

more of the experimental units have the same response or 40% or more have one of only two response 

values or 65% or more have one of only three response values. An exact version of the JonckheereTerpstra test is used for the step-down trend test and an exact version of the Mann-Whitney test with 

Bonferroni-Holm adjusted p-values is used in place of Dunn’s test. 

3.2 Comparisons of the F0 and F1 generations 

 Mysid data for the two generation multi-concentration test were analyzed according to a common 

procedure for each generation as described above. Analyses were also carried out compare the results in 

the F0 and F1 generations. Analyses were carried out using data reported on a per mysid, per breeding 

pair, or per basket basis. The comparisons of the F0 and F1 generations were performed with a two way 

mixed effects ANOVA model, which included generation, concentration group, and the interaction of 

generation and concentration as fixed effects and replicate (concentration, generation) as a random effect, 

allowing unequal variances between the two generations. The residual variance was treated as 

homogeneous or heterogeneous between generations basen on the results of Levene’s test. 

 Preliminary tests were carried out to detect outliers, departures from normality, and heterogeneity 

of variance. These analyses were independent of those carried out for the analyses within generations and 

did not necessarily arrive at the same conclusions. Subsequent comparisons among groups were carried 

out using procedures based on the outcome of the preliminary tests. All test procedures described in this 

report are based on the default auto option programs in STATCHARRMS 

3.2.1 Outlier screens 

In STATCHARMS, the outliers for the comparisons of two generations were based on the twoway mixed effect model in section 3.2.6.Outliers were detected based on the residuals from the mixed 

effects model fit using Tukey’s outlier rule on the untransformed data. Any response more than 1.5 times 

the interquartile range above the third quartile (75th percentile) or below the first quartile (25th percentile) 

is considered an outlier by Tukey’s rule. These outliers were documented together with those identified in 

the one generation analyses and will be reported to EPA for review. EPA will determine if the deviations, 

although statistically significant, are in fact biologically important. EPA will determine whether the 

flagged values should be included in the subsequent analyses or should be treated as outliers. The subsets 

of potential outliers are response specific. Due to time constraints, the subsequent summaries and 
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statistical analyses in this report were carried out including the flagged values. The list of screened 

values is presented in the STATACHARRMS output. 

3.2.2 Normality 

 Normality was formally assessed in the mixed effects ANOVA procedure using the Shapiro-Wilk 

test (Shapiro and Wilk, 1965). It was assessed based on residuals from the 2-way mixed effects ANOVA 

model fit, where group and generation as well as group by generation interaction were fixed effects. The 

random replicate tank-to-tank variance was assumed by STATCHARRMS to be heterogeneous between 

generations. Residual variance within replicate tanks was assumed to be homogeneous between 

generations. Screened potential outliers were included in the tests. 

3.2.3 Transformation of data 

 There are three options for data transformation in STATCHARRMS: log-transformation, square 

root transformation, and arc-sin square-root transformation. Under the auto option in STATCHARRMS, 

the analysis procedures are first applied to the untransformed data. If the normality assumption is valid, 

then no transformation is necessary. If the normality assumption is invalid, then normality tests are again 

performed on the log and the square-root transformed data. If the normality assumption is valid after a 

transformation, then the procedures are applied to the transformed data, including the homogeneity of 

variance analysis. If the normality assumption is invalid for both transformations, then nonparametric 

procedures are applied. Transformation is then not necessary as any transformation does not change the 

rank scores for the data when the nonparametric procedures are used. 

3.2.4 Heterogeneous variances 

 If the normality assumption was accepted for either the untransformed or the transformed data, 

heterogeneity of variances among the generations and the concentration groups was assessed using the 

ANOVA procedure and Levene’s test with absolute residuals (Levene, 1960), where the residuals were 

from the model fit described in the normality test (Section 3.2.2). Outliers were included in the test. 

3.2.5 Monotonicity Test 

 A test for monotonicity was not carried out as ordered alternatives procedures were not available 

in the two way mixed effects ANOVA in STATCHARRMS. 

3.2.6 F0 generation versus F1 generation 

 If the normality assumptions held, a parametric two-way mixed effects analysis was applied to 

the untransformed or transformed data (whichever satisfied the normality assumption). Otherwise, a 

nonparametric mixed effects analysis was applied to the rank scores. If the generation main effect or the 

interaction of concentration by generation effect was significant, F-tests for significant differences 

between generations, within concentration groups (including the control group), the so-called slices in the 

ANOVA table, were carried out to identify the concentration groups for which the two generations were 

significantly different. 
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4. RESULTS 

 For each endpoint the summary tables present the following statistical analysis for the control 

group and each concentration group for each generation: 

 Number of samples 

 Mean 

 Standard deviation 

 CV (%) 

These summary statistics were calculated across all samples and all baskets for each concentration group 

and for each generation. Summary results for the two generations are presented in Tables 1A and 1B 

(reproduction endpoints), Tables 2A and 2B (growth endpoints), Tables 3A and 3B (survival endpoints). 

These means and standard deviations are also displayed, sided-by-side for the two generations in Figures 

1 through 22. The summaries and figures are based on untransformed data with no screened outliers 

removed. 

 Statistical analysis was performed according to the statistical methods described in the methods 

section using STATCHARRMS. The results of statistical analyses by generation are summarized in 

Tables 1C and 1D, Tables 2C and 2D, and Tables 3C and 3D. Comparisons between the F0 and F1 

generations are summarized in Tables 1E, 2E and 3E.
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Table 1A. Summary of Reproduction Endpoints. Laboratory: UNT, Test Chemical: Vinclozolin, Generation: F

Concentration Statistics 

Time to 

Mature 

(Females) 

Number 

Offspring 

(Brood 1) 

Number 

Offspring 

(Brood 2) 

Number 

Offspring 

(Two 

Broods) 

Days to 

Brood 1 

Days to 

Brood 2 

Days 

Between 

Broods 1 

and 2 

% of Females 

Productively 

Active 

Sex Ratio 

(males/females) 

Control 

(Group 1) 

Mean 11.333 3.800 4.900 8.700 17.800 25.412 7.588 95.233 1.472 

Standard 

Dev. 

0.577 2.191 3.432 4.692 2.745 3.759 3.429 8.256 0.284 

N 3 20 20 20 20 17 17 3 3 

CV (%) 5.09 57.66 70.04 53.93 15.42 14.79 45.19 8.67 19.31 

25 ppb 

(Group 2) 

Mean 12.000 4.714 4.400 8.933 17.750 24.769 6.769 95.233 1.241 

Standard 

Dev. 

1.000 2.217 3.680 4.559 2.049 2.522 2.166 8.256 0.409 

N 3 21 15 15 20 13 13 3 3 

CV(%) 8.33 47.02 83.64 51.03 11.54 10.18 32.00 8.67 32.98 

50 ppb 

(Group 3) 

Mean 10.000 3.000 3.714 6.714 19.389 24.182 5.455 85.700 1.336 

Standard 

Dev. 

0.000 1.495 3.024 2.525 3.775 3.311 2.382 14.300 0.463 

N 3 18 14 14 18 11 11 3 3 

CV(%) 0.00 49.84 81.41 37.60 19.47 13.69 43.67 16.69 34.62 

100 ppb 

(Group 4) 

Mean 10.333 5.095 3.533 9.133 18.571 23.000 5.625 100.000 2.248 

Standard 

Dev. 

1.155 3.506 4.373 4.051 3.310 2.268 1.768 0.000 0.801 

N 3 21 15 15 21 8 8 3 3 

CV(%) 11.17 68.80 123.77 44.35 17.82 9.86 31.43 0.00 35.62 

200 ppb 

(Group 5) 

Mean 9.667 4.789 5.400 10.067 18.789 26.214 7.357 90.467 2.354 

Standard 

Dev. 

0.577 1.653 3.460 4.234 2.394 3.093 2.818 8.256 0.781 

N 3 19 15 15 19 14 14 3 3 

CV(%) 5.97 34.50 64.07 42.06 12.74 11.80 38.30 9.13 33.17 

600 ppb 

(Group 6) 

Mean 10.667 2.769 3.400 6.100 21.000 26.200 5.200 65.567 3.922 

Standard 

Dev. 

0.577 1.787 3.658 4.864 4.000 3.493 0.447 15.032 0.958 

N 3 13 10 10 11 5 5 3 3 

CV(%) 5.41 64.52 107.58 79.73 19.05 13.33 8.60 22.93 24.43 

1. Summary statistics are calculated across all replicates and all samples in the entire concentration group. 

2. Time to mature, % of females productively active, sex ratio are measured per tank (replicate), Number of offspring, day to broods, days between broods are measur3. Summaries are based on untransformed data with no screened outliers removed . 
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15 

Table 1B. Summary of Reproduction Endpoints. Laboratory: UNT, Test Chemical: Vinclozolin, Generation: F

Concentration Statistics 

Time to Mature 

(Females) 

Number 

Offspring 

(Brood 1) 

Number 

Offspring 

(Brood 2) 

Number 

Offspring 

(Two Broods) 

Days to 

Brood 1 

Days to 

Brood 2 Days Between Broods 1 and 2 

% 

Pr

Control 

(Group 1) 

Mean 9.333 4.143 7.200 11.350 16.095 21.150 5.000 

Standard Dev. 2.082 1.931 2.419 3.117 0.700 0.671 0.324 

N 3 21 20 20 21 20 20 

CV(%) 22.30 46.61 33.60 27.46 4.35 3.17 6.49 

25 ppb 

(Group 2) 

Mean 12.000 5.286 6.889 12.000 18.619 23.625 5.375 

Standard Dev. 2.828 2.148 3.359 4.256 4.177 4.380 1.310 

N 2 21 18 18 21 16 16 

CV(%) 23.57 40.64 48.76 35.47 22.43 18.54 24.38 

50 ppb 

(Group 3) 

Mean 11.000 5.500 7.944 13.556 18.200 23.111 5.000 

Standard Dev. 2.000 2.439 3.038 4.655 3.037 3.234 0.485 

N 3 20 18 18 20 18 18 

CV(%) 18.18 44.34 38.25 34.34 16.68 13.99 9.70 

100 ppb 

(Group 4) 

Mean 8.333 4.381 5.474 9.684 16.524 21.353 4.882 

Standard Dev. 1.155 2.312 3.272 3.400 1.289 1.412 0.485 

N 3 21 19 19 21 17 17 

CV(%) 13.86 52.79 59.78 35.11 7.80 6.61 9.94 

200 ppb 

(Group 5) 

Mean 10.500 5.800 6.333 12.111 17.600 22.625 6.125 

Standard Dev. 0.707 4.417 4.387 6.716 3.169 2.973 2.100 

N 2 10 9 9 10 8 8 

CV(%) 6.73 76.16 69.28 55.46 18.01 13.14 34.29 

600 ppb 

(Group 6) 

Mean 8.333 5.000 3.750 8.800 17.619 25.000 7.000 

Standard Dev. 2.517 2.258 3.307 3.365 2.037 3.588 3.000 

N 3 21 20 20 21 17 17 

CV(%) 30.20 45.17 88.18 38.24 11.56 14.35 42.86 

1. Summary statistics are calculated across all replicates and all samples in the entire concentration group. 

2. Time to mature, % of females productively active, sex ratio are measured per tank (replicate), Number of offspring, day to broods, days between broods are 3. Summaries are based on untransformed data with no screened outliers removed . 
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Table 1C. Statistical Analysis Results for Reproduction Endpoints. Laboratory: UNT, Test 

 Chemical: Vinclozolin, Generation: F0 

Parameters 

Normality 

Test1

Homogeneous 

of Variance 

Test2

Monotonicity 

Test3

Concentration 

Trend Tests4

Overall 

Concentration 

Effect5

Pairwise 

Comparisons 

to Control6,7

Time to Mature Normal Homogeneous Monotonic No Yes NA 

Number 

Offspring 

(Brood 1)

NonNormal NA NonMonotonic NA Yes No 

Number 

Offspring 

(Brood 2)

NonNormal NA NonMonotonic NA No NA 

Number 

Offspring 

(Two Broods)

Normal Homogeneous NonMonotonic NA No NA 

Days to 

Brood 1 

NonNormal NA Monotonic 

Increasing trend 

for the two 

highest dose 

groups 

Yes NA 

Days to 

Brood 2 Normal Homogeneous NonMonotonic NA No NA 

Days Between 

Broods 1 and 2 

NonNormal NA NonMonotonic NA No NA 

% of Females 

Productively 

Active 

NonNormal NA Monotonic 

Decreasing trend 

for the highest 

dose group 

Yes NA 

Sex Ratio Normal Homogeneous NonMonotonic NA Yes Group 6 

1. Shapiro-Wilk test for normality. 

2. Levene Test for Homogeneity of Variances. 

3. Monotonicity is tested using linear and quadratic contrasts under one-way ANOVA as applied to normalized rank statistics. 

4. Concentration trends are tested using a step-down Jonckheere-Terpstra test procedure. 

5. If normality assumption is valid one way ANOVA is applied to the data and F-test is used to determine the concentration effect. If 

normality assumption is invalid, Kruskal-Wallis test is applied to test for the overall concentration effect. 

6. Control is group 1 and test concentration groups are group 2 to 6. 

7. If overall concentration of ANOVA is significant and monotonicity is rejected, comparisons of test concentration to the control are 

conducted: 

a. using Dunnett’s test with homogeneous within-treatment variances and using Tamhane-Dunnett test with heterogeneous 

within-treatment variances for normal distributed data 

b. using a modified (nonparametric) Dunn’s test if data are not normally distributed. 
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Table 1D. Statistical Analysis Results for Reproduction Endpoints. Laboratory: UNT, Test 

 Chemical: Vinclozolin, Generation: F1 

Parameters 

Normality 

Test1

Homogeneous 

of Variance 

Test2

Monotonicity 

Test3

Concentration 

Trend Tests4

Overall 

Concentration 

Effect5

Pairwise 

Comparisons 

to Control6,7

Time to Mature Normal Homogeneous NonMonotonic NA No NA 

Number Offspring 

(Brood 1) Normal8 Homogeneous NonMonotonic NA No NA 

Number Offspring 

(Brood 2) Normal Homogeneous Monotonic Decreasing trend for 

the highest dose group Yes NA 

Number Offspring 

(Two Broods) Normal9 Homogeneous Monotonic Decreasing trend for 

the highest dose group Yes NA 

Days to Brood 1 NonNormal NA NonMonotonic NA No NA 

Days to Brood 2 NonNormal NA Monotonic Increasing trend for 

the highest dose group Yes NA 

Days Between Broods 1 

and 2 Normal NA NonMonotonic NA Yes Group 6 

% of Females 

Reproductively Active 

NonNormal NA NonMonotonic NA No NA 

Sex Ratio Normal Homogeneous NonMonotonic NA Yes No 

1. Shapiro-Wilk test for normality. 

2. Levene Test for Homogeneity of Variances. 

3. Monotonicity is tested using linear and quadratic contrasts under one-way ANOVA as applied to normalized rank statistics. 

4. Concentration Trends are tested using a step-down Jonckheere-Terpstra test procedure. 

5. If normality assumption is valid one way ANOVA is applied to the data and F-test is used to determine the concentration effect. If 

normality assumption is invalid, Kruskal-Wallis test is applied to test for the overall concentration effect. 

6. Control is group 1 and test concentration groups are group 2 to 6. 

7. If overall concentration of ANOVA is significant and monotonicity is rejected, comparisons of test concentration to the control are 

conducted: 

a. using Dunnett’s test with homogeneous within-treatment variances and using Tamhane-Dunnett test with heterogeneous 

within-treatment variances for normal distributed data 

b. using a modified (nonparametric) Dunn’s test if data are not normally distributed. 

8. Normal when square root-transformed. 

9. Normal when log-transformed. 

 
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Table 1E. Comparisons between Two Generations for Reproduction Endpoints. Laboratory: 

 UNT, Test Chemical: Vinclozolin. 

Parameters 

Normality 

Test1

Homogeneous 

of Variance 

Test2

Parameteric/ 

Nonparametric 

Concentration 

Main Effect 

Generation Main 

Effect 

Concentration 

by Generation 

Interaction 

Groups with 

Significant 

Difference 

between 

Generations3

Time to 

Mature Normal Homogeneous Parametric No No No NA 

Number 

Offspring 

(Brood 1

Normal Homogeneous Parametric No Yes No Groups 3 and 

6 

Number 

Offspring 

(Brood 2)

Normal Homogeneous Parametric No Yes No Group 3 

Number 

Offspring 

(Two Broods)

Normal Homogeneous Parametric No Yes No Groups 2 and 

3 

Days to 

Brood 1 

NonNormal NA Nonparametric Yes No No NA 

Days to 

Brood 2 Normal Homogeneous Parametric Yes Yes No Group 5 

Days Between 

Broods 1 and 

2 

NonNormal NA Nonparametric Yes No No NA 

% of Females 

Productively 

Active 

NonNormal NA Nonparametric Yes No Yes Group 6 

Sex Ratio Normal Homogeneous Parametric Yes Yes Yes Group 4, 5 

and 6 

1. Shapiro-Wilk test for normality. 

2. Levene test for homogeneous of variances. 

3. Control is group 1 and test concentration groups are group 2 to 6. 

4. . 

 
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Table 2A. Summary of Growth Endpoints. Laboratory: UNT, Test Chemical: Vinclozolin, 

 Generation: F01,2,3,4,5

Concentration Statistics 

Dry 

Weight (mg) 

(day 0) 

Dry Weight 

(mg) 

(day 7) 

Dry Weight 

(mg) 

(day 14) 

Dry Weight 

(mg) 

(Termination) 

Length 

(mm) 

(day 0) 

Length 

(mm) 

(day 7) 

Length 

(mm) 

(day 14) 

Length 

(mm) 

(Termination) 

Control 

(Group 1) 

Mean 0.033 0.200 0.522 1.043 1.641 3.572 4.914 6.225 

Standard Dev. 0.004 0.030 0.132 0.266 0.189 0.364 0.384 0.410 

N 7 9 36 32 34 18 36 32 

CV(%) 13.55 15.21 25.38 25.53 11.53 10.20 7.82 6.58 

25 ppb 

(Group 2) 

Mean 0.175 0.565 0.904 3.489 5.119 6.218 

Standard Dev. 0.049 0.181 0.180 0.395 0.565 0.347 

N 9 31 34 18 31 34 

CV(%) 28.07 32.02 19.90 11.33 11.03 5.58 

50 ppb 

(Group 3) 

Mean 0.191 0.541 0.953 3.567 5.015 6.039 

Standard Dev. 0.085 0.104 0.202 0.376 0.249 0.346 

N 9 34 27 18 34 28 

CV(%) 44.43 19.17 21.19 10.53 4.96 5.72 

100 ppb 

(Group 4) 

Mean 0.186 0.487 0.944 3.528 5.179 6.241 

Standard Dev. 0.030 0.166 0.228 0.276 0.598 0.359 

N 9 24 39 18 24 39 

CV(%) 15.99 34.10 24.11 7.83 11.54 5.75 

200 ppb 

(Group 5) 

Mean 0.194 0.476 0.885 3.456 5.080 6.230 

Standard Dev. 0.112 0.133 0.235 0.419 0.574 0.370 

N 9 25 34 18 25 33 

CV(%) 57.52 27.96 26.51 12.13 11.31 5.93 

600 ppb 

(Group 6) 

Mean 0.158 0.386 0.837 3.344 4.726 6.182 

Standard Dev. 0.047 0.111 0.173 0.479 0.450 0.438 

N 9 23 28 18 23 28 

CV(%) 29.62 28.80 20.65 14.33 9.53 7.09 

1. Summary statistics are calculated across all replicates and all samples in the entire concentration group. 

2. On day 0, all mysids are in control environments. 

3. Length is measured per mysid. Weight were measured on averages per (replicate) tank on day 0, per mating pair on day 7, and per mysid 

on day 14 and termination. 

4. Summaries are based on untransformed data with no screened outliers removed. 

5. Replicate tank within group information is not provided for the growth endpoints. 

 
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Table 2B. Summary of Growth Endpoints. Laboratory: UNT, Test Chemical: Vinclozolin, 

 Generation: F11,2,3,4

Concentration Statistics 

Dry 

Weight (mg) 

(day 0) 

Dry Weight 

(mg) 

(day 14) 

Dry Weight 

(mg) 

(Termination) 

Length 

(mm) 

(day 0) 

Length 

(mm) 

(day 14) 

Length 

(mm) 

(Termination) 

Control 

(Group 1) 

Mean 0.058 0.602 0.931 1.340 5.276 6.100 

Standard Dev. 0.050 0.095 0.201 0.123 0.199 0.283 

N 9 29 34 45 29 34 

CV(%) 87.42 15.73 21.57 9.20 3.78 4.64 

25 ppb 

(Group 2) 

Mean 0.044 0.574 1.037 1.349 5.292 5.933 

Standard Dev. 0.049 0.097 0.251 0.155 0.215 0.405 

N 9 12 40 45 12 40 

CV(%) 110.41 16.86 24.23 11.47 4.07 6.82 

50 ppb 

(Group 3) 

Mean 0.064 0.581 1.021 1.402 5.252 6.358 

Standard Dev. 0.042 0.133 0.235 0.144 0.198 0.302 

N 9 25 39 45 25 38 

CV(%) 65.88 22.83 23.00 10.25 3.77 4.75 

100 ppb 

(Group 4) 

Mean 0.050 0.585 1.041 1.391 5.206 6.292 

Standard Dev. 0.053 0.114 0.289 0.135 0.223 0.383 

N 9 32 37 45 32 36 

CV(%) 105.88 19.57 27.79 9.67 4.28 6.08 

200 ppb 

(Group 5) 

Mean 0.066 0.654 1.064 1.369 5.200 6.357 

Standard Dev. 0.033 0.056 0.269 0.116 0.312 0.287 

N 9 8 13 45 8 14 

CV(%) 50.43 8.49 25.28 8.51 5.99 4.52 

600 ppb 

(Group 6) 

Mean 0.068 0.509 1.070 1.402 5.282 6.598 

Standard Dev. 0.036 0.114 0.317 0.133 0.429 0.431 

N 8 22 42 40 22 42 

CV(%) 53.42 22.33 29.63 9.48 8.13 6.53 

1. Summary statistics are calculated across all replicates and all samples in the entire concentration group. 

2. Length is measured per mysid. Weight were measured on averages per (replicate) tank on day 0, per mating pair on day 7, and per mysid 

on day 14 and termination. 

3. Summaries are based on untransformed data with no screened outliers removed. 

4. Replicate tank within group information is not provided for the growth endpoints. 

 
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Table 2C. Statistical Analysis Results for Reproduction Endpoints. Laboratory: UNT, Test 

 Chemical: Vinclozolin, Generation: F0 

Parameters 

Normality 

Test1

Homogeneous 

of Variance 

Test2

Monotonicity 

Test3

Concentration 

Trend Tests4

Overall 

Concentration 

Effect5

Pairwise 

Comparisons 

to Control6,7

Dry 

Weight(mg) 

 (day 7)

Normal8 Homogeneous NonMonotonic NA No NA 

Dry 

Weight(mg) 

(day 14)

Normal Heterogeneous NonMonotonic NA Yes Group 6 

Dry Weight 

(mg) 

(Termination)

Normal Homogeneous Monotonic 

Decreasing trend 

for the highest 

dose group 

Yes NA 

Length (mm) 

(day 7) Normal Homogeneous NonMonotonic NA No NA 

Length (mm) 

(day 14)

NonNormal NA NonMonotonic NA Yes Groups 2 and 

4 

Length (mm) 

(Termination)

NonNormal NA NonMonotonic NA No NA 

1. Shapiro-Wilk test for normality. 

2. Levene Test for Homogeneity of Variances. 

3. Monotonicity is tested using linear and quadratic contrasts under one-way ANOVA as applied to normalized rank statistics. 

4. Concentration Trends are tested using a step-down Jonckheere-Terpstra test procedure. 

5. If normality assumption is valid one way ANOVA is applied to the data and F-test is used to determine the concentration effect. If 

normality assumption is invalid, Kruskal-Wallis test is applied to test for the overall concentration effect. 

6. Control is group 1 and test concentration groups are group 2 to 6. 

7. If overall concentration of ANOVA is significant and monotonicity is rejected, comparisons of test concentration to the control are 

conducted: 

a. using Dunnett’s test with homogeneous within-treatment variances and using Tamhane-Dunnett test with heterogeneous withintreatment variances for normal distributed data 

b. using a modified (nonparametric) Dunn’s test if data are not normally distributed. 

8. Normal when square root-transformed. 

 
DRAFT 

Summary Tables and Figures for Mysid: UNT Vinclozolin Data 

22 

Table 2D. Statistical Analysis Results for Reproduction Endpoints. Laboratory: UNT, Test 

 Chemical: Vinclozolin, Generation: F1 

Parameters 

Normality 

Test1

Homogeneous 

of Variance 

Test2

Monotonicity 

Test3

Concentration 

Trend Tests4

Overall 

Concentration 

Effect5

Pairwise 

Comparisons 

to Control6,7

Dry Weight 

(mg) 

(day 0)

Normal8 Homogeneous NonMonotonic NA No NA 

Dry Weight 

(mg) 

(day 14)

Normal8 Homogeneous NonMonotonic NA Yes Group 6 

Dry Weight 

(mg) 

(Termination)

NonNormal NA NonMonotonic NA No NA 

Length (mm) 

(day 0)

NonNormal NA Monotonic 

Increasing trend 

for the highest 

dose group 

Yes NA 

Length (mm) 

(day 14) Normal Heterogeneous NonMonotonic NA Yes No 

Length (mm) 

(Termination) Normal Homogeneous Monotonic 

Increasing trend 

for the highest 

four dose groups 

Yes NA 

1. Shapiro-Wilk test for normality. 

2. Levene Test for homogeneous of variances. 

3. Monotonicity is tested using linear and quadratic contrasts under one-way ANOVA as applied to normalized rank statistics. 

4. Concentration trends are tested using a step down Jonckheere-Terpstra test procedure. 

5. If normality assumption is valid one way ANOVA is applied to the data and F-test is used to determine the concentration effect. If 

normality assumption is invalid, Kruskal-Wallis test is applied to test for the overall concentration effect. 

6. Control is group 1 and test concentration groups are group 2 to 6. 

7. If overall concentration of ANOVA is significant and monotonicity is rejected, comparisons of test concentration to the control are 

conducted: 

a. using Dunnett’s test with homogeneous within-treatment variances and using Tamhane-Dunnett test with heterogeneous 

within-treatment variances for normal distributed data 

b. using a modified (nonparametric) Dunn’s test if data are not normally distributed. 

8. Normal when log-transformed. 

 
DRAFT 

Summary Tables and Figures for Mysid: UNT Vinclozolin Data 

23 

Table 2E. Comparisons between Two Generations for Growth Endpoints. Laboratory: UNT, 

 Test Chemical: Vinclozolin. 

Parameters 

Normality 

Test1

Homogeneous 

of Variance 

Test2

Parameteric/ 

Nonparametric 

Concentration 

Main Effect 

Generation 

Main Effect 

Concentration 

by Generation 

Interaction 

Groups with 

Significant 

Difference 

between 

Generations3

Dry Weight 

(mg) 

(day 14)

Normal Homogeneous Parametric No Yes No Groups 4, 5 

and 6 

Dry Weight 

(mg) 

(Termination)

Normal Homogeneous Parametric No Yes No Group 6 

Length (mm) 

(day 14) Normal Homogeneous Parametric No Yes No Group 6 

Length (mm) 

(Termination) Normal Homogeneous Parametric Yes No Yes Groups 2, 3 

and 6 

1. Shapiro-Wilk test for normality. 

2. Levene test for homogeneous of variances. 

3. Control is group 1 and test concentration groups are group 2 to 6. 

 
DRAFT 

Summary Tables and Figures for Mysid: UNT Vinclozolin Data 

24 

Table 3A. Summary of Survival Endpoints. Laboratory: UNT, Test Chemical: Vinclozolin, 

 Generation: F01,2,3

Concentration Statistics 

Reproduction 

Survival Rate (%) 

(day 7) 

Reproduction 

Survival Rate (%) 

(day 14) 

Reproduction 

Survival Rate (%) 

(Termination) 

Total 

Survival 

Rate (%) 

(day 7) 

Total 

Survival Rate 

(%) (day 14) 

Control 

(Group 1) 

Mean 100.000 85.000 72.309 100.000 88.235 

Standard 

Dev. 0.000 5.000 7.647 0.000 2.941

N 3 3 3 3 3

CV(%) 0 5.88 10.58 0 3.33

25 ppb 

(Group 2) 

Mean 98.333 76.667 70.476 95.833 78.431 

Standard 

Dev. 2.887 12.583 3.434 3.819 10.329

N 3 3 3 3 3

CV(%) 2.94 16.41 4.87 3.98 13.17

50 ppb 

(Group 3) 

Mean 93.333 80.000 64.583 95.833 84.314 

Standard 

Dev. 7.638 8.660 9.547 2.887 3.396

N 3 3 3 3 3

CV(%) 8.18 10.83 14.78 3.01 4.03

100 ppb 

(Group 4) 

Mean 96.667 90.000 67.479 97.500 83.333 

Standard 

Dev. 2.887 13.229 12.953 2.500 6.123

N 3 3 3 3 3

CV(%) 2.99 14.7 19.2 2.56 7.35

200 ppb 

(Group 5) 

Mean 93.333 85.000 65.926 94.167 82.353 

Standard 

Dev. 7.638 8.660 18.802 5.204 5.094

N 3 3 3 3 3

CV(%) 8.18 10.19 28.52 5.53 6.19

600 ppb 

(Group 6) 

Mean 88.333 73.333 67.857 90.000 70.588 

Standard 

Dev. 2.887 7.638 9.449 5.000 2.941

N 3 3 3 3 3

CV(%) 3.27 10.41 13.93 5.56 4.17

1. Summary statistics are calculated across all replicates and all samples in the entire concentration group. 

2. Survival rates were measured per tank. 

3. Summaries are based on untransformed data with no screened outliers removed. 

 
DRAFT 

Summary Tables and Figures for Mysid: UNT Vinclozolin Data 

25 

Table 3B. Summary of Survival Endpoints. Laboratory: UNT, Test Chemical: Vinclozolin, 

 Generation: F11,2,3

Concentration Statistics 

Reproduction 

Survival Rate (%) 

(day 7) 

Reproduction 

Survival Rate (%) 

(day 14) 

Reproduction 

Survival Rate (%) 

(Termination) 

Total 

Survival 

Rate (%) 

(day 7) 

Total 

Survival Rate 

(%) (day 14) 

Control 

(Group 1) 

Mean 97.778 93.333 72.639 96.444 80.889 

Standard 

Dev. 3.849 6.667 19.792 3.355 22.562

N 3 3 3 3 3

CV(%) 3.94 7.14 27.25 3.48 27.89

25 ppb 

(Group 2) 

Mean 86.667 84.444 73.849 86.923 82.479 

Standard 

Dev. 11.547 13.878 11.616 11.331 16.732

N 3 3 3 3 3

CV(%) 13.32 16.43 15.73 13.04 20.29

50 ppb 

(Group 3) 

Mean 100.000 97.778 79.779 96.154 91.197 

Standard 

Dev. 0.000 3.849 17.536 3.846 4.737

N 3 3 3 3 3

CV(%) 0 3.94 21.98 4 5.19

100 ppb 

(Group 4) 

Mean 97.778 95.556 83.929 98.889 96.420 

Standard 

Dev. 3.849 3.849 11.152 1.925 3.710

N 3 3 3 3 3

CV(%) 3.94 4.03 13.29 1.95 3.85

200 ppb 

(Group 5) 

Mean 40.000 22.222 63.889 43.333 31.778 

Standard 

Dev. 40.000 38.490 19.642 16.667 5.004

N 3 3 2 3 3

CV(%) 100 173.21 30.74 38.46 15.75

600 ppb 

(Group 6) 

Mean 86.349 53.016 83.194 91.904 60.716 

Standard 

Dev. 13.345 46.329 9.320 8.757 41.074

N 3 3 3 3 3

CV(%) 15.45 87.39 11.2 9.53 67.65

1. Summary statistics are calculated across all replicates and all samples in the entire concentration group. 

2. Survival rates were measured per tank. 

3. Summaries are based on untransformed data with no screened outliers removed. 
DRAFT 

Summary Tables and Figures for Mysid: UNT Vinclozolin Data 

26 

Table 3C. Statistical Analysis Results for Reproduction Endpoints. Laboratory: UNT, Test 

 Chemical: Vinclozolin, Generation: F0 

Parameters Normality 

Test1

Homogeneous 

of Variance 

Test2

Monotonicity 

Test3

Concentration 

Trend Tests4

Overall 

Concentration 

Effect5

Pairwise 

Comparisons 

to Control6,7

Reproduction 

Survival Rate 

(%) 

(day 7)

Normal Homogeneous Monotonic 

Decreasing trend 

for the highest 

dose group 

No NA 

Reproduction 

Survival Rate 

(%) 

(day 14)

Normal Homogeneous NonMonotonic NA No NA 

Reproduction 

Survival Rate 

(%) 

(Termination)

Normal Homogeneous NonMonotonic NA No NA 

Total 

Survival Rate 

(%) 

(day 7)

Normal Homogeneous Monotonic 

Decreasing trend 

for the highest 

dose group 

No NA 

Total 

Survival Rate 

(%) (day 14)

Normal Homogeneous Monotonic 

Decreasing trend 

for the highest 

dose group 

Yes NA 

1. Shapiro-Wilk test for normality. 

2. Levene Test for homogeneous of variances. 

3. Monotonicity is tested using linear and quadratic contrasts under one-way ANOVA as applied to normalized rank statistics. 

4. Concentration trends are tested using a step down Jonckheere-Terpstra test procedure. 

5. If normality assumption is valid one way ANOVA is applied to the data and F-test is used to determine the concentration effect. If 

normality assumption is invalid, Kruskal-Wallis test is applied to test for the overall concentration effect. 

6. Control is group 1 and test concentration groups are group 2 to 6. 

7. If overall concentration of ANOVA is significant and monotonicity is rejected, comparisons of test concentration to the control are 

conducted: 

a. using Dunnett’s test with homogeneous within-treatment variances and using Tamhane-Dunnett test with heterogeneous 

within-treatment variances for normal distributed data 

b. using a modified (nonparametric) Dunn’s test if data are not normally distributed. 
DRAFT 

Summary Tables and Figures for Mysid: UNT Vinclozolin Data 

27 

Table 3D. Statistical Analysis Results for Reproduction Endpoints. Laboratory: UNT, Test 

 Chemical: Vinclozolin, Generation: F1 

Parameters 

Normality 

Test1

Homogeneous 

of Variance 

Test2

Monotonicity 

Test3

Concentration 

Trend Tests4

Overall 

Concentration 

Effect5

Significant 

Pairwise 

Comparisons 

to Control6,7

Reproduction 

Survival Rate 

(%) 

(day 7)

NonNormal NA NonMonotonic NA No NA 

Reproduction 

Survival Rate 

(%) 

(day 14)

Normal Homogeneous Monotonic 

Decreasing trend 

for the highest 

dose group 

Yes NA 

Reproduction 

Survival Rate 

(%) 

(Termination)

Normal Homogeneous NonMonotonic NA No NA 

Total 

Survival Rate 

(%) 

(day 7)

Normal Homogeneous Monotonic No Yes NA 

Total 

Survival Rate 

(%) (day 14)

Normal Homogeneous NonMonotonic NA Yes Group 5 

1. Shapiro-Wilk test for normality. 

2. Levene Test for homogeneous of variances. 

3. Monotonicity is tested using linear and quadratic contrasts under one-way ANOVA as applied to normalized rank statistics. 

4. Concentration trends are tested using a step down Jonckheere-Terpstra test procedure. 

5. If normality assumption is valid one way ANOVA is applied to the data and F-test is used to determine the concentration effect. If 

normality assumption is invalid, Kruskal-Wallis test is applied to test for the overall concentration effect. 

6. Control is group 1 and test concentration groups are group 2 to 6. 

7. If overall concentration of ANOVA is significant and monotonicity is rejected, comparisons of test concentration to the control are 

conducted: 

a. using Dunnett’s test with homogeneous within-treatment variances and using Tamhane-Dunnett test with heterogeneous 

within-treatment variances for normal distributed data 

b. using a modified (nonparametric) Dunn’s test if data are not normally distributed. 

 
DRAFT 

Summary Tables and Figures for Mysid: UNT Vinclozolin Data 

28 

Table 3E. Comparisons between Two Generation Survival Endpoints. Laboratory: UNT, Test 

 Chemical: Vinclozolin. 

Parameters 

Normality 

Test1

Homogeneous 

of Variance 

Test2

Parameteric/ 

Nonparametric 

Concentration 

Main Effect 

Generation 

Main Effect 

Concentration 

by Generation 

Interaction 

Groups with 

Significant 

Difference 

between 

Generations3

Reproduction 

Survival Rate 

(%) 

(day 7)

NonNormal NA Nonparametric Yes No No NA 

Reproduction 

Survival Rate 

(%) 

(day 14)

Normal Homogeneous Parametric Yes No Yes Group 5 

Reproduction 

Survival Rate 

(%) 

(Termination)

Normal Homogeneous Parametric No No No NA 

Total 

Survival Rate 

(%) 

(day 7)

Normal Homogeneous Parametric Yes Yes Yes Group 5 

Total 

Survival Rate 

(%) (day 14)

Normal Homogeneous Parametric Yes No Yes Group 5 

1. Shapiro-Wilk test for normality. 

2. Levene test for homogeneous of variances. 

3. Control is group 1 and test concentration groups are group 2 to 6. 
DRAFT 

Summary Tables and Figures for Mysid: UNT Vinclozolin Data 

29 

Figure 1. Time to Mature By Generation and Group For Chemical Vinclozolin From Lab 

UNT. 

Figure 2. Number Offspring (brood 1) By Generation and Group For Chemical Vinclozolin 

From Lab UNT. 

Time To Mature (Days)

6

7

8

9

10

11

12

13

14

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Group

Generation F0 Generation F1

Number Offspring (brood 1)

1

2

3

4

5

6

7

8

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Group

Generation F0 Generation F1
DRAFT 

Summary Tables and Figures for Mysid: UNT Vinclozolin Data 

30 

Figure 3. Number Offspring (brood 2) By Generation and Group For Chemical Vinclozolin 

From Lab UNT. 

Figure 4. Number Offspring (two broods) By Generation and Group For Chemical 

Vinclozolin From Lab UNT. 

Number Offspring (brood 2)

1

2

3

4

5

6

7

8

9

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Group

Generation F0 Generation F1

Number Offspring (two broods)

3

4

5

6

7

8

9

10

11

12

13

14

15

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Group

Generation F0 Generation F1
DRAFT 

Summary Tables and Figures for Mysid: UNT Vinclozolin Data 

31 

Figure 5. Days to Brood 1 By Generation and Group For Chemical Vinclozolin From Lab 

UNT. 

Figure 6. Days to Brood 2 By Generation and Group For Chemical Vinclozolin From Lab 

UNT. 

Days to Brood 1

15

16

17

18

19

20

21

22

23

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Group

Generation F0 Generation F1

Days to Brood 2

21

22

23

24

25

26

27

28

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Group

Generation F0 Generation F1
DRAFT 

Summary Tables and Figures for Mysid: UNT Vinclozolin Data 

32 

Figure 7. Days Between Broods 1 and 2 By Generation and Group For Chemical Vinclozolin 

From Lab UNT. 

Figure 8. Percent of Females Productively Active By Generation and Group For Chemical 

Vinclozolin From Lab UNT. 

Days Between Broods 1 and 2

4

5

6

7

8

9

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Group

Generation F0 Generation F1

% of Females Productively Active

50

60

70

80

90

100

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Group

Generation F0 Generation F1
DRAFT 

Summary Tables and Figures for Mysid: UNT Vinclozolin Data 

33 

Figure 9. Sex Ratio By Generation and Group For Chemical Vinclozolin From Lab UNT. 

Figure 10. Dry Weight (mg) (day 0) By Generation and Group For Chemical Vinclozolin From 

Lab UNT. 

Sex Ratio

0

1

2

3

4

5

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Group

Generation F0 Generation F1

Dry Weight (mg)(day 0)

0.02

0.03

0.04

0.05

0.06

0.07

0.08

0.09

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Group

Generation F0 Generation F1
DRAFT 

Summary Tables and Figures for Mysid: UNT Vinclozolin Data 

34 

Figure 11. Dry Weight (mg) (day 7) By Generation and Group For Chemical Vinclozolin From 

Lab UNT. 

Figure 12. Dry Weight (mg) (day 14) By Generation and Group For Chemical Vinclozolin 

From Lab UNT. 

Dry Weight (mg)(day 7)

0.14

0.15

0.16

0.17

0.18

0.19

0.20

0.21

0.22

0.23

0.24

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Group

Generation F0 Generation F1

Dry Weight (mg)(day 14)

0.35

0.40

0.45

0.50

0.55

0.60

0.65

0.70

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Group

Generation F0 Generation F1
DRAFT 

Summary Tables and Figures for Mysid: UNT Vinclozolin Data 

35 

Figure 13. Dry Weight (mg) (Termination) By Generation and Group For Chemical 

Vinclozolin From Lab UNT. 

Figure 14. Length (mm) (day 0) By Generation and Group For Chemical Vinclozolin From 

Lab UNT. 

Dry Weight (mg)(Termination)

0.80

0.85

0.90

0.95

1.00

1.05

1.10

1.15

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Group

GenerationF0 GenerationF1

Length (mm)(day 0)

1.30

1.35

1.40

1.45

1.50

1.55

1.60

1.65

1.70

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Group

Generation F0 Generation F1
DRAFT 

Summary Tables and Figures for Mysid: UNT Vinclozolin Data 

36 

Figure 15. Length (mm) (day 7) By Generation and Group For Chemical Vinclozolin From 

Lab UNT. 

Figure 16. Length (mm) (day 14) By Generation and Group For Chemical Vinclozolin From 

Lab UNT. 

Length (mm)(day 7)

3.20

3.25

3.30

3.35

3.40

3.45

3.50

3.55

3.60

3.65

3.70

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Group

Generation F0 Generation F1

Length (mm)(day 14)

4.6

4.7

4.8

4.9

5.0

5.1

5.2

5.3

5.4

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Group

Generation F0 Generation F1
DRAFT 

Summary Tables and Figures for Mysid: UNT Vinclozolin Data 

37 

Figure 17. Length (mm) (Termination) By Generation and Group For Chemical Vinclozolin 

From Lab UNT. 

Figure 18. Reproduction Survival Rate (%) (day 7) By Generation and Group For Chemical 

Vinclozolin From Lab UNT. 

Length (mm)(Termination)

5.8

5.9

6.0

6.1

6.2

6.3

6.4

6.5

6.6

6.7

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Group

GenerationF0 GenerationF1

Reproduction Survival Rate (%)(day 7)

10

20

30

40

50

60

70

80

90

100

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Group

Generation F0 Generation F1
DRAFT 

Summary Tables and Figures for Mysid: UNT Vinclozolin Data 

38 

Figure 19. Reproduction Survival Rate (%) (day 14) By Generation and Group For Chemical 

Vinclozolin From Lab UNT. 

Figure 20. Reproduction Survival Rate (%) (Termination) By Generation and Group For 

Chemical Vinclozolin From Lab UNT. 

Reproduction Survival Rate (%)(day 14)

0

10

20

30

40

50

60

70

80

90

100

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Group

Generation F0 Generation F1

Reproduction Survival Rate (%)(Termination)

30

40

50

60

70

80

90

100

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Group

Generation F0 Generation F1
DRAFT 

Summary Tables and Figures for Mysid: UNT Vinclozolin Data 

39 

Figure 21. Total Survival Rate (%) (day 7) By Generation and Group For Chemical 

Vinclozolin From Lab UNT. 

Figure 22. Total Survival Rate (%) (day 14) By Generation and Group For Chemical 

Vinclozolin From Lab UNT. 

Total Survival Rate (%)(day 7)

20

30

40

50

60

70

80

90

100

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Group

Generation F0 Generation F1

Total Survival Rate (%)(day 14)

10

20

30

40

50

60

70

80

90

100

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Control 25

ppb

50

ppb

100

ppb

200

ppb

600

ppb

Group

Generation F0 Generation F1