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Intra-Laboratory Analysis Report for Mysid: WLI Octylphenol Data 

Endocrine Disruptor Screening Program 

Contract No: EP-W-11-063 

Task Order No: 2 

Maintain the Data Coordination Center 

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 WLI Octylphenol 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 .................................................................................................................... 7 

3.1 Comparisons of test chemical with the control ......................................................................... 7 

3.1.1 Outlier Screens ...................................................................................................................... 7 

3.1.2 Normality .............................................................................................................................. 8 

3.1.3 Transformation of data .......................................................................................................... 8 

3.1.4 Heterogeneous variances ....................................................................................................... 8 

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 ............................................................................. 10 

4. Results ..................................................................................................................................... 11 

Tables

Table 0. Data information for SMV Octylphenol (Only control data were available for 

F1 generation)............................................................................................................................ 4 

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

Octylphenol, Generation: F01,2 ................................................................................................ 12 

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

Octylphenol, Generation: F11,2,3 .............................................................................................. 13 

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

Chemical: Octylphenol, Generation: F0 .................................................................................. 14 

Table 2A. Summary of Growth Endpoints. Laboratory: WLI, Test Chemical: Octylphenol, 

Generation: F01,2,3 .................................................................................................................... 15 

Table 2B. Summary of Growth Endpoints. Laboratory: WLI, Test Chemical: Octylphenol, 

Generation: F11,2,3 .................................................................................................................... 16 

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

Chemical: Octylphenol, Generation: F0 .................................................................................. 17 
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Table 3A. Summary of Survival Endpoints. Laboratory: WLI, Test Chemical: Octylphenol, 

Generation: F01,2 ...................................................................................................................... 18 

Table 3B. Summary of Survival Endpoints. Laboratory: WLI, Test Chemical: Octylphenol, 

Generation: F11,2 ...................................................................................................................... 19 

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

Chemical: Octylphenol, Generation: F0 .................................................................................. 20 

Figures

Figure 1. Time to Mature for Generation F0 by Group for WLI Lab, Chemical Octylphenol. .............. 21 

Figure 2. Number Offspring (Brood 1) for Generation F0 by Group for WLI Lab, Chemical 

Octylphenol. ............................................................................................................................ 21 

Figure 3. Number Offspring (Brood 2) for Generation F0 by Group for WLI Lab, Chemical 

Octylphenol. ............................................................................................................................ 22 

Figure 4. Number Offspring (Two Broods) for Generation F0 by Group for WLI Lab, Chemical 

Octylphenol. ............................................................................................................................ 22 

Figure 5. Days to Brood 1 for Generation F0 by Group for WLI Lab, Chemical Octylphenol. ............. 23 

Figure 6. Days to Brood 2 for Generation F0 by Group for WLI Lab, Chemical Octylphenol. ............. 23 

Figure 7. Days Between Broods 1 and 2 for Generation F0 by Group for WLI Lab, Chemical 

Octylphenol. ............................................................................................................................ 24 

Figure 8. Percent of Females Productively Active for Generation F0 by Group for WLI Lab, Chemical 

Octylphenol. ............................................................................................................................ 24 

Figure 9. Sex Ratio for Generation F0 by Group for WLI Lab, Chemical Octylphenol. ....................... 25 

Figure 10. Dry Weight (mg) (day 0) for Generation F0 by Group for WLI Lab, Chemical 

Octylphenol. ............................................................................................................................ 25 

Figure 11. Dry Weight (mg) (day 7) for Generation F0 by Group for WLI Lab, Chemical 

Octylphenol. ............................................................................................................................ 26 

Figure 12. Dry Weight (mg) (day 14) for Generation F0 by Group for WLI Lab, Chemical 

Octylphenol ............................................................................................................................ .26 

Figure 13. Dry Weight (mg) (Termination) for Generation F0 by Group for WLI Lab, Chemical 

Octylphenol. ............................................................................................................................ 27 

Figure 14. Length (mm) (day 0) for Generation F0 by Group for WLI Lab, Chemical Octylphenol. ..... 27 

Figure 15. Length (mm) (day 7) for Generation F0 by Group for WLI Lab, Chemical Octylphenol. ..... 28 

Figure 16. Length (mm) (day 14) for Generation F0 by Group for WLI Lab, Chemical Octylphenol. ... 28 

Figure 17. Length (mm) (Termination) for Generation F0 by Group for WLI Lab, Chemical 

Octylphenol. ............................................................................................................................ 29 
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Figure 18. Reproduction Juvenile Survival Rate - Day 6 (%) for Generation F0 by Group for WLI Lab, 

Chemical Octylphenol. ............................................................................................................ 29 

Figure 19. Reproduction Juvenile Survival Rate - Day 14 (%) for Generation F0 by Group for WLI Lab, 

Chemical Octylphenol. ............................................................................................................ 30 

Figure 20. Total Juvenile Survival Rate - Day 6 (%) for Generation F0 by Group for WLI Lab, Chemical 

Octylphenol. ............................................................................................................................ 30 

Figure 21. Total Juvenile Survival Rate - Day 14 (%) for Generation F0 by Group for WLI Lab, 

Chemical Octylphenol. ............................................................................................................ 31 

Figure 22. Reproduction Adult Survival Rate at Termination (%) for Generation F0 by Group for WLI 

Lab, Chemical Octylphenol ..................................................................................................... 31 

 
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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. Two laboratories, SMV and WLI, conducted two-generation mysid survival and growth tests 

following a similar protocol with the test chemical octylphenol. Statistical analyses were carried out on 

the SMV and WLI octylphenol study data. The draft analysis documented in this report is based on the 

WLI octylphenol test data. Statistical analysis of the octylphenol effects in the WLI study is being carried 

out with the F0 generation data. The reproductive output from the F0 generation mysids treated with 

octylphenol in this study was not sufficient to produce an F1 generation. Only the negative control 

replicates were carried to the F1 generation and are included in the F1 portion of the summarization and 

analysis. 

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

that are grown to sexual 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 

sexual maturity and their offspring are collected (F1 offspring). The F0 test is conducted with five graded 

concentrations octylphenol 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. In this study only the control mysids were carried to the F1 

generation 

 The F1 generation (control mysids only) is begun with newly hatched F0 control offspring. 

Within each replicate tank the growth and reproduction baskets each initially include 15 newly hatched 

mysids, or the maximum available. 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 control group brood cups and transferred immediately to 

baskets with untreated negative control water. 

 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 
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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 46 for WLI 

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

 For the intra-laboratory analysis of the WLI octylphenol mysid study comparisons were made 

between the F0 test chemical concentration groups and the concurrent F0 control group to assess the 

mysid responses to graded concentrations of octylphenol. The F1 generation control data were 

summarized but were not statistically analyzed. 

2. STUDY DATA 

2.1 Data Endpoints 

The test method specifies 25 endpoints. Some of the endpoints are specified per mysid or 

composite of mysids (e.g. day 0 weight, day 7 weight), some are specified per breeding pair, and 

some are specified per replicate tank per basket (i.e. per replicate tank). Not all of the specified 

responses were determined in the WLI octylphenol study (e.g. number of reproductive days per 

female. 

These endpoints were divided among three categories: 

(i) Reproduction (F0) (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) 

(ii) Survival (F0) (6) 

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

survival) 

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

at days 7, 14 and termination (total survival) 

 
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 The survival rates were adjusted for the number of F0 mysids that were removed for 

weight and length determinations (i.e. were omitted from both the numerator and denominator) 

and for other non-treatment related deaths 

(iii) Growth (F0) (8)1 

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 5 mysids) 

f. Weight at day 7 (on average per 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. 

Growth 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 growth basket mysids on day 14), and for the surviving reproduction 

mysids at termination. For the weight data on day 0, there were a total of 9 composites (5 mysids 

per composite). For day 7 there were 3 composite weights per replicate (2 mysids per 

composite) and three replicates per concentration group. 

 Data were a available for F1 generation for the above parameters for the control group 

only. 

 The test laboratory ran a concurrent control and five targeted test concentration levels for 

the F0 generation. 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. 

 

1

 Day 14 and termination weights and lengths were reported by gender. 
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Table 0. Data information for SMV Octylphenol (Only control data were available for F1 

generation)

Data Files and data sheets Endpoints Comments 

Original- 

 Mys Repro Data.xls 

− Repro Spreadsheet 

 Mys Repro Data F1.xls 

− Repro Spreadsheet 

 Mys Repro Time.xls 

these information are available in the 

Mys Repro Data.xls, Thus this data 

does not need to be read in 

 Sex ration Sas.xls 

there are two data sheets SASdata (2) 

and SASdata. They seems not the 

same. For now we will use SASdata 

(2) 

 Growth & Survival F1.xls 

(last datasheet for F1 sex ratio); 

Reformatted-: 

 WLI Octylphenol Reproduction 

F0.xlsx 

 WLI Octylphenol Reproduction 

F0.xlsx 

 WLI Octylphenol Sex Ratio F0.xlsx 

 WLI Octylphenol Sex Ratio F1.xlsx 

 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 endpoints 

are not available: 

 Offspring per female per 

reproductive day for the first 

two broods 

 Total reproductively active 

days for the first two broods 

 If there are no offspring for a 

brood. The data are recorded as 

zero for the brood sizes. Per 

EPA suggestion, such data are 

changed as: zero if female is 

still living, and “NA” (recorded 

as missing) for a brood (1st or 

2nd brood) if female died 

without releasing the brood. 

 The total offspring for the first 

2 broods are therefore 

recalculated and displayed in 

the reformatted data. 

 Time to maturity is same for all 

animals as “14”. 

 %of female reproductively 

active, sex ratio are reported per 

tank. The other reproductive 

endpoints are reported per 

mating pair. 

Original- 

 Growth & Survival.xls

 Growth & Survival F1.xls 

The following extra data files provided 

the same information included in the 

above two files: 

 Day 0 Weight and lengths Data.xls 

(it look like that this is F0 data); 

 Day 0 Lengths Raw Data.xls (it is 

control data, it looks like F1 data. 

It actually include both lengths and 

weights) ; 

 Day 7 Weights and lengths Raw 

Data.xls (F0 data); 

 Day 7 Growth Measurements Raw 

Data F1.xls; 

 Day 14 Weights and lengths Raw 

Data.xls (F0 data); 

 Day 14 Growth Measurements 

Raw Data.xls (F1 data); 

 Term Weights and lengths.xls 

(termination for F0) 

 Mysid-Term Growth Raw Data NC 

Rep. A.xls (termination for F1: 

Weight (dry weight) and length for 

days 0, 7, 14 and termination 

 For F0 generation all mysids on 

day 0 are kept in untreated 

water. There were 9 

composites (5 mysids per 

composite). 

 Length are reported per mysid. 

 Weight is reported per 2 mysid 

composites for day 7, and per 

mysid for day 14 and for 

termination. 

 Genders of mysids are provided 

for day 14 and termination. 
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Data Files and data sheets Endpoints Comments 

control replicate A, the next two 

files for replicates B and C); 

 Mysid-Term Growth Raw 

Data_NCB.xls; 

 Mysid-Term Growth Raw 

Data_NCC.xls 

Reformatted- 

 WLI Octylphenol Growth 

F0_Created From SAS.xlsx 

 WLI Octylphenol Growth 

F1_Export From SAS.xlsx 

Original- 

 Growth & Survival.xls

− Juvenile Survival-Day 6 

− Juvenile Survival-Day 7-Pair 

− Adult Survival 

 Growth & Survival F1.xls 

Reformatted- SMV Octylphenol 

Survival.xls:

- F0ReproductionSurvival; 

- F0GrowthSurvival 

- F0TotalSurvival 

- F1ReproductionSurvival 

- F1GrowthSurvival 

- F1TotalSurvival 

 For F0, Survival 

(reproduction) (day 6 before 

pair, survival on day 14) 

 For F0, Survival (total) (day 6 

before pair, survival on day 

14),reproductive adult 

survival at termination. 

 For F1, Survival 

(reproduction) (day 7, day 14, 

and at termination) – controls 

only 

 For F1, Survival (total) (day 7, 

day 14) – controls only 

 For F0 generation: 

− Data sheet “Juvenile SurvivalDay 6”. We labeled them as 

Juvenile Survival (Day 6). Both 

reproductive survival and total 

survival are given in this data 

sheet. 

− Sheet “Juvenile Survival-Day 

7-Pair” has title l “Juvenile 

Survival Day 7 - Pairing on Day 

14”. We labeled them as 

“Juvenile Survival (Day 7)”. It 

is actually survival on day 14. 

Both reproductive survival and 

total survival are given in this 

worksheet. 

 Worksheet “Adult Survival” 

has title “total adult survival 

after pairing”. It is in fact 

reproduction survival at 

termination. There is a column 

called “Total Adult Survival 

After Pairing Excluding 

Impinged Mysids”, which are 

not in the other two survival 

sheets. It is in fact reproduction 

termination survival excluding 

nontoxic related mortalities.. 

 For the F1 generation: 

− Worksheet “Juvenile SurvivalDay 6 (2)” is for Survival on 

Day 7. 

− Juvenile Survival-Day14 (2) is 

for day7 to 14. 

− Both reproductive survival and 

total survival are given on these 

two data sheets. 

− Sheet “Adult Survival” has title 

“total adult survival after 
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Data Files and data sheets Endpoints Comments 

pairing”. It is in fact 

reproductive survival for day 14 

to termination. There is a 

column called “Total Adult 

Survival After Pairing 

Excluding Impinged Mysids”. 

It is in fact reproduction 

termination survival excluding 

nontoxic related mortalities. 

 We have changed the denominators in the survival rate calculations for day 14 and 

termination to reflect total survival from day 0, adjusting for the nontoxic related mortality. The 

survival rates presented in this report differ from those included in the WLI spreadsheet because 

they represent survival from day 0. 

2.3 Analysis Considerations 

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

 Since only F0 generation data are available for the intra-laboratory analysis, the 

STATCHARRMS one-way ANOVA procedure is used. In the one-way ANOVA 

STATCHARRMS pools the results across replicate tanks within test or control groups. 

The one-way ANOVA (parametric or nonparametric form) was applied to the one 

generation data and there are no random replicate effects. The within generation analysis 

compares the test group results to the control group results. 

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

were carried out independently for the analyses within the F0 generation only. 

 Reproductive response was observed for 46 days, which is much longer than the other 

laboratory studies (around 30 days). It might have increased the opportunity to observe 

the second broods for some mysids. Termination survival is also at 46 days. This may 

impact the survival rates. 

 Survival is calculated by WLI differently than the other laboratories. Survival rates are 

presented in the WLI spreadsheet at day 6, between days 7 and 14, and between days 14 

and termination. The denominators of the survival rates are the numbers of mysids 

surviving to the beginnings of the survival periods. This has been modified in this report 

to represent survival from day 0. 

 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 zero value in the analysis. EPA 

has suggested that if the female is living the 0 brood size should be counted as 0. If the 

female died before the recorded “0” brood size is generated, the brood size should be 

counted as missing. Battelle has changed the data entries in the columns for brood1 size 

and brood2 size in the Excel spreadsheet according to this suggestion. In the reformatted 
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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 zero or missing, the times for the 

brood was also recorded as “-“ 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 is also treated as missing. 

 In the WLI octylphenol spreadsheets the 14 day and termination length and weight 

responses are presented by gender. In the preliminary draft analyses the 14 day and 

termination weights and lengths are pooled across male and female mysids. 

 There is no information about number of reproductive days per female or number of 

offspring per reproductive day per female. These endpoints are not included in the 

analysis in this report. 

3. STATISTICAL METHODS 

Statistical analysis procedures used conform to the analysis procedures in the 

STATCHARRMS package. Statistical analyses compare the test chemical responses with the 

control responses within the F0 generation. There is no analysis of the F1 generation. The 

statistical analyses were carried out in a univariate fashion, independently for each endpoint. 

There is no comparison of the F0 and F1 generations. 

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 procedure were applied to per mysid or per mysid-pair data without first 

averaging across the mysids in the same the replicate baskets. 

3.1 Comparisons of test chemical with the control 

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 the STATCHARRMS 

3.1.1 Outlier Screens 

 Outlier screens 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 
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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 were to 

be documented and reported to EPA for review to 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 statistical 

summaries and analyses in this draft report were carried out including the flagged values. The 

list of screened values is including in the STATACHARRMS outputs and will be sent to EPA. 

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. 
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3.1.5 Monotonicity of response 

 In STATCHARRMS, a monotonicity test using linear and quadratic contrasts under oneway ANOVA was applied to normalized rank statistics (to avoid the complications that can arise 

from non-normal 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: 

 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 alternatives section for more details. 
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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 or 

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 responses to the control responses were conducted using a modified Dunn’s test. 

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 non-parametric 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 

 Comparisons of the F0 and F1 generation were not carried out as only the F0 generation 

was available for the intra-laboratory analysis. 
DRAFT 

Intra-Laboratory Analysis Report for Mysid: WLI Octylphenol Data 

11 

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. Summary results for the F0 generation are presented in Table 1A 

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

These means and standard deviations are also displayed in Figures 1 through 22. Summary 

results for the control group in the F1 generation are presented in Table 1B (reproduction 

endpoints), Table 2B (growth endpoints), and Table 3B (survival endpoints). 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 for the F0 

generation are summarized in Table 1C (reproduction), Table 2C (growth), and Table 3C 

(survival). 
DRAFT 

Intra-Laboratory Analysis Report for Mysid: WLI Octylphenol Data 

12 

Table 1A. Summary of Reproduction Endpoints. Laboratory: WLI, Test Chemical: Octylphenol, Generation: 

Dose 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 Fem

Producti

Active 

Control 

(Group 1) 

Mean 14.000 3.471 5.800 9.067 26.250 34.077 9.000 76.167 

Standard Dev. 0.000 2.375 4.554 5.007 5.871 6.251 4.243 8.256 

N 21 17 15 15 16 13 13 3 

CV (%) 0 68.43 78.52 55.22 22.37 18.34 47.14 10.84 

0.26 μg/L (Group 2) 

Mean 14.000 3.150 2.167 5.278 26.778 33.222 9.000 85.700 

Standard Dev. 0.000 2.498 2.834 4.099 6.656 5.167 4.062 14.300 

N 21 20 18 18 18 9 9 3 

CV(%) 0 79.3 130.78 77.66 24.86 15.55 45.13 16.69 

0.64 μg/L (Group 3) 

Mean 14.000 3.211 1.722 4.944 28.938 35.100 7.900 76.167 

Standard Dev. 0.000 2.507 2.347 3.796 7.362 7.094 4.841 8.256 

N 21 19 18 18 16 10 10 3 

CV(%) 0 78.1 136.25 76.77 25.44 20.21 61.28 10.84 

1.6 μg/L (Group 4) 

Mean 14.000 2.300 1.222 3.667 26.438 38.167 10.000 71.400 

Standard Dev. 0.000 2.452 2.045 3.498 13.038 6.735 5.550 14.300 

N 21 20 18 18 16 6 6 3 

CV(%) 0 106.59 167.34 95.4 49.32 17.65 55.5 20.03 

4.0 μg/L (Group 5) 

Mean 14.000 2.056 0.500 2.556 30.357 35.000 6.667 66.633 

Standard Dev. 0.000 2.508 1.465 2.935 7.880 7.937 2.887 8.256 

N 21 18 18 18 14 3 3 3 

CV(%) 0 122.02 293.06 114.85 25.96 22.68 43.3 12.39 

10 μg/L (Group 6) 

Mean 14.000 3.867 3.100 7.200 25.800 36.000 9.000 71.400 

Standard Dev. 0.000 3.701 3.604 5.653 5.226 4.690 4.050 14.300 

N 21 15 10 10 15 6 6 3 

CV(%) 0 95.71 116.26 78.51 20.26 13.03 45 20.03 

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.
DRAFT 

Intra-Laboratory Analysis Report for Mysid: WLI Octylphenol Data 

13 

Table 1B. Summary of Reproduction Endpoints. Laboratory: WLI, Test Chemical: Octylphenol, Generation: 

Dose 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 Fem

Producti

Active 

Control 

(Group 1) 

Mean 14.000 1.364 1.818 4.364 19.286 25.250 6.500 61.66

Standard Dev. 0.000 1.433 2.994 5.732 4.716 4.573 3.000 12.58

N 11 11 11 11 7 4 4 3 

CV (%) 0 105.11 164.67 131.36 24.45 18.11 46.15 20.4 

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 m3. Only control data were available for F1 generation.
DRAFT 

Intra-Laboratory Analysis Report for Mysid: WLI Octylphenol Data 

14 

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

Chemical: Octylphenol, 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 NA NA No NA No NA 

Number 

Offspring 

(Brood 1)

Normal8 Homogeneous NonMonotonic NA No NA 

Number 

Offspring 

(Brood 2)

NonNormal NA NonMonotonic NA Yes Group 2, 3, 4 

& 5 

Number 

Offspring 

(Two Broods)

Normal8 Homogeneous NonMonotonic NA Yes Group 4 & 5 

Days to 

Brood 1 

NonNormal NA NonMonotonic NA No NA 

Days to 

Brood 2 Normal Homogeneous NonMonotonic NA No NA 

Days Between 

Broods 1 and 2 Normal9 Homogeneous NonMonotonic NA No NA 

% of Females 

Productively 

Active 

Normal Homogeneous NonMonotonic NA No NA 

Sex Ratio Normal Homogeneous NonMonotonic NA No NA 

1. Shapiro-Wilk test for normality. 

2. LeveneTest 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. 
DRAFT 

Intra-Laboratory Analysis Report for Mysid: WLI Octylphenol Data 

15 

Table 2A. Summary of Growth Endpoints. Laboratory: WLI, Test Chemical: Octylphenol, Generation: F01,2,3

Dose 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) 

Le

(m

(da

Control 

(Group 1) 

Mean 0.045 0.597 0.662 1.407 1.916 5.550 5

Standard Dev. 0.010 0.062 0.102 0.241 0.111 0.368 0

N 9 9 41 32 45 18 

CV(%) 22.6 10.33 15.4 17.15 5.78 6.63 4

0.26 μg/L 

(Group 2) 

Mean 0.508 0.598 1.462 5.083 6

Standard Dev. 0.042 0.086 0.240 0.257 0

N 9 40 39 18 

CV(%) 8.28 14.4 16.43 5.06 6

0.64 μg/L 

 (Group 3) 

Mean 0.488 0.643 1.400 5.094 6

Standard Dev. 0.118 0.107 0.240 0.472 0

N 9 38 28 17 

CV(%) 24.23 16.64 17.12 9.27 4

1.6 μg/L 

(Group 4) 

Mean 0.600 0.624 1.427 5.233 6

Standard Dev. 0.073 0.107 0.266 0.270 0

N 9 40 36 18 

CV(%) 12.13 17.08 18.62 5.16 6

4.0 μg/L 

(Group 5) 

Mean 0.533 0.590 1.394 4.911 6

Standard Dev. 0.095 0.114 0.234 0.478 0

N 9 41 33 18 

CV(%) 17.84 19.39 16.81 9.72 6

10 μg/L 

(Group 6) 

Mean 0.518 0.560 1.326 4.989 6

Standard Dev. 0.087 0.098 0.333 0.459 0

N 9 39 26 18 

CV(%) 16.72 17.49 25.14 9.2 5

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 terminatio
DRAFT 

Intra-Laboratory Analysis Report for Mysid: WLI Octylphenol Data 

16 

Table 2B. Summary of Growth Endpoints. Laboratory: WLI, Test Chemical: Octylphenol, Generation: F11,2,3

Dose 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) 

L

(

(d

Control 

(Group 1) 

Mean 0.044 0.340 0.847 1.394 1.889 5.200 8

Standard Dev. 0.011 0.045 0.111 0.217 0.129 0.486 0

N 4 6 15 20 19 12 

CV(%) 24.72 13.15 13.12 15.56 6.81 9.35 

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 term

3. Only control data were available for F1 generation. 
DRAFT 

Intra-Laboratory Analysis Report for Mysid: WLI Octylphenol Data 

17 

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

Chemical: Octylphenol, 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) Normal Homogeneous Non-Monotonic NA Yes Group 3 

Dry Weight(mg) 

(day 14) Normal Homogeneous Monotonic 

Decreasing trend for 

the two highest dose 

groups 

Yes NA 

Dry Weight (mg) 

(Termination) Non-Normal NA Non-Monotonic NA No NA 

Length (mm) 

(day 7) Non-Normal NA Monotonic 

Decreasing trend for 

the two highest dose 

groups 

Yes NA 

Length (mm) 

(day 14) Non-Normal NA Non-Monotonic NA Yes Groups 2, 3, 4, 5 & 

6 

Length (mm) 

(Termination) Non-Normal NA Non-Monotonic NA No NA 

1. Shapiro-Wilk test for normality. 

2. LeveneTest 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. 

 
DRAFT 

Intra-Laboratory Analysis Report for Mysid: WLI Octylphenol Data 

18 

Table 3A. Summary of Survival Endpoints. Laboratory: WLI, Test Chemical: Octylphenol, 

Generation: F01,2

Dose 

Concentration Statistics 

Reproduction 

Juvenile 

Survival rate (%) 

(day 6) 

Reproduction 

Juvenile 

Survival rate (%) 

(day 14) 

Total 

Juvenile 

Survival rate (%) 

(day 6) 

Total 

Juvenile 

Survival rate (%) 

(day 14) 

Reproductive 

Adult 

Survival rate (%) 

(Termination) 

Control 

(Group 1) 

Mean 100.000 91.667 100.000 95.098 69.583

Standard 

Dev. 

0.000 7.638 0.000 4.493 3.411

N 3 3 3 3 3

CV(%) 0 8.33 0 4.72 4.9

0.26 μg/L 

(Group 2) 

Mean 98.333 96.667 99.167 96.078 82.185

Standard 

Dev. 

2.887 5.774 1.443 3.396 8.791

N 3 3 3 3 3

CV(%) 2.94 5.97 1.46 3.53 10.7

0.64 μg/L 

 (Group 3) 

Mean 100.000 86.667 100.000 90.196 57.778

Standard 

Dev. 

0.000 14.434 0.000 9.455 10.184

N 3 3 3 3 3

CV(%) 0 16.65 0 10.48 17.63

1.6 μg/L 

(Group 4) 

Mean 100.000 95.000 99.167 96.078 75.000

Standard 

Dev. 

0.000 5.000 1.443 1.698 10.825

N 3 3 3 3 3

CV(%) 0 5.26 1.46 1.77 14.43

4.0 μg/L 

(Group 5) 

Mean 100.000 98.333 100.000 98.039 74.644

Standard 

Dev. 

0.000 2.887 0.000 3.396 4.707

N 3 3 3 3 3

CV(%) 0 2.94 0 3.46 6.31

10 μg/L 

(Group 6) 

Mean 96.667 95.000 97.500 94.118 60.513

Standard 

Dev. 

2.887 0.000 0.000 2.941 13.363

N 3 3 3 3 3

CV(%) 2.99 0 0 3.12 22.08

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

2. Survival rates were measured per tank. 

 
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Intra-Laboratory Analysis Report for Mysid: WLI Octylphenol Data 

19 

Table 3B. Summary of Survival Endpoints. Laboratory: WLI, Test Chemical: Octylphenol, 

Generation: F11,2

Dose 

Concentration Statistics 

Reproduction 

Juvenile 

Survival rate 

(%) (day 6) 

Reproduction 

Juvenile 

Survival rate 

(%) (day 14) 

Total 

Juvenile 

Survival rate 

(%) 

(day 6) 

Total 

Juvenile 

Survival rate 

(%) 

(day 14) 

Reproductive 

Adult 

Survival rate 

(%) 

(Termination) 

Control 

(Group 1) 

Mean 88.533 86.325 89.103 90.103 61.640 

Standard 

Dev. 

7.407 6.161 7.997 2.156 14.425 

N 3 3 3 3 3 

CV(%) 8.37 7.14 8.98 2.39 23.4 

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

2. Survival rates were measured per tank. 

3. Only control data were available for F1 generation. 

 
DRAFT 

Intra-Laboratory Analysis Report for Mysid: WLI Octylphenol Data 

20 

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

Chemical: Octylphenol, Generation: F0 

Parameters Normality 

Test1

Homogeneous of 

Variance Test2

Monotonicity 

Test3

Concentration Trend 

Tests4

Overall 

Concentration 

Effect5

Pairwise 

comparisons to 

Control6,7

Reproduction 

Juvenile 

Survival Rate (%) 

(day 6)

Non-Normal NA Non-Monotonic NA No NA 

Reproduction 

Juvenile 

Survival Rate (%) 

(day 14)

Normal Homogeneous Non-Monotonic NA No NA 

Total 

Juvenile 

Survival Rate (%) 

(day 6)

Non-Normal NA Monotonic No No NA 

Total 

Juvenile 

Survival Rate (%) 

(day 14)

Normal Homogeneous Non-Monotonic NA No NA 

Reproductive Adult 

Survival rate (%) 

(Termination)

Normal Homogeneous Non-Monotonic NA No 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 

Intra-Laboratory Analysis Report for Mysid: WLI Octylphenol Data 

21 

Figure 1. Time to Mature for Generation F0 by Group for WLI Lab, Chemical Octylphenol. 

Figure 2. Number Offspring (Brood 1) for Generation F0 by Group for WLI Lab, Chemical 

Octylphenol. 

Time To Mature (Days)

13

14

15

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0

Number Offspring (brood 1)

1

2

3

4

5

6

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0
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Intra-Laboratory Analysis Report for Mysid: WLI Octylphenol Data 

22 

Figure 3. Number Offspring (Brood 2) for Generation F0 by Group for WLI Lab, Chemical 

Octylphenol. 

Figure 4. Number Offspring (Two Broods) for Generation F0 by Group for WLI Lab, 

Chemical Octylphenol. 

Number Offspring (brood 2)

0

1

2

3

4

5

6

7

8

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0

Number Offspring (two broods)

0

2

4

6

8

10

12

14

16

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0
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Intra-Laboratory Analysis Report for Mysid: WLI Octylphenol Data 

23 

Figure 5. Days to Brood 1 for Generation F0 by Group for WLI Lab, Chemical Octylphenol. 

Figure 6. Days to Brood 2 for Generation F0 by Group for WLI Lab, Chemical Octylphenol. Days to Brood 1

20

22

24

26

28

30

32

34

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0

Days to Brood 2

28

30

32

34

36

38

40

42

44

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0
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Intra-Laboratory Analysis Report for Mysid: WLI Octylphenol Data 

24 

Figure 7. Days Between Broods 1 and 2 for Generation F0 by Group for WLI Lab, Chemical 

Octylphenol. 

Figure 8. Percent of Females Productively Active for Generation F0 by Group for WLI Lab, 

Chemical Octylphenol. 

Days Between Broods 1 and 2

4

6

8

10

12

14

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0

% of Females Productively Active

60

65

70

75

80

85

90

95

100

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0
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Intra-Laboratory Analysis Report for Mysid: WLI Octylphenol Data 

25 

Figure 9. Sex Ratio for Generation F0 by Group for WLI Lab, Chemical Octylphenol. 

Figure 10. Dry Weight (mg) (day 0) for Generation F0 by Group for WLI Lab, Chemical 

Octylphenol. 

Sex Ratio

0.6

0.8

1.0

1.2

1.4

1.6

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0

Dry Weight (mg)(day 0)

0.040

0.041

0.042

0.043

0.044

0.045

0.046

0.047

0.048

0.049

0.050

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0
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Intra-Laboratory Analysis Report for Mysid: WLI Octylphenol Data 

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Figure 11. Dry Weight (mg) (day 7) for Generation F0 by Group for WLI Lab, Chemical 

Octylphenol. 

Figure 12. Dry Weight (mg) (day 14) for Generation F0 by Group for WLI Lab, Chemical 

Octylphenol. 

Dry Weight (mg)(day 7)

0.40

0.42

0.44

0.46

0.48

0.50

0.52

0.54

0.56

0.58

0.60

0.62

0.64

0.66

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0

Dry Weight (mg)(day 14)

0.50

0.52

0.54

0.56

0.58

0.60

0.62

0.64

0.66

0.68

0.70

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0
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Figure 13. Dry Weight (mg) (Termination) for Generation F0 by Group for WLI Lab, 

Chemical Octylphenol. 

Figure 14. Length (mm) (day 0) for Generation F0 by Group for WLI Lab, Chemical 

Octylphenol. 

Dry Weight (mg)(Termination)

1.20

1.25

1.30

1.35

1.40

1.45

1.50

1.55

1.60

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0

Length (mm)(day 0)

1.89

1.90

1.91

1.92

1.93

1.94

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0
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Intra-Laboratory Analysis Report for Mysid: WLI Octylphenol Data 

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Figure 15. Length (mm) (day 7) for Generation F0 by Group for WLI Lab, Chemical 

Octylphenol. 

Figure 16. Length (mm) (day 14) for Generation F0 by Group for WLI Lab, Chemical 

Octylphenol. 

Length (mm)(day 7)

4.6

4.8

5.0

5.2

5.4

5.6

5.8

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0

Length (mm)(day 14)

5.7

5.8

5.9

6.0

6.1

6.2

6.3

6.4

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0
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Figure 17. Length (mm) (Termination) for Generation F0 by Group for WLI Lab, Chemical 

Octylphenol. 

Figure 18. Reproduction Juvenile Survival Rate - Day 6 (%) for Generation F0 by Group for 

WLI Lab, Chemical Octylphenol. 

Length (mm)(Termination)

8.7

8.8

8.9

9.0

9.1

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0

Reproduction Juvenile Survival Rate - Day 6 (%)

94

95

96

97

98

99

100

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0
DRAFT 

Intra-Laboratory Analysis Report for Mysid: WLI Octylphenol Data 

30 

Figure 19. Reproduction Juvenile Survival Rate - Day 14 (%) for Generation F0 by Group for 

WLI Lab, Chemical Octylphenol. 

Figure 20. Total Juvenile Survival Rate - Day 6 (%) for Generation F0 by Group for WLI Lab, 

Chemical Octylphenol. 

Reproduction Juvenile Survival Rate - Day 14 (%)

75

80

85

90

95

100

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L 4ug/L

10

ug/L

Group

Generation F0

Total Juvenile Survival Rate - Day 6 (%)

97.0

97.5

98.0

98.5

99.0

99.5

100.0

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L 4ug/L

10

ug/L

Group

Generation F0
DRAFT 

Intra-Laboratory Analysis Report for Mysid: WLI Octylphenol Data 

31 

Figure 21. Total Juvenile Survival Rate - Day 14 (%) for Generation F0 by Group for WLI 

Lab, Chemical Octylphenol. 

Figure 22. Reproduction Adult Survival Rate at Termination (%) for Generation F0 by Group 

for WLI Lab, Chemical Octylphenol 

Total Juvenile Survival Rate - Day 14 (%)

80

82

84

86

88

90

92

94

96

98

100

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0

Reproductive Adult Survival Rate (%) (Termination) 50

55

60

65

70

75

80

85

90

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0