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Intra-Laboratory Analysis Report for Mysid: SMV 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 SMV 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 ............................................................................................................ 5 

3. Statistical Methods ................................................................................................................................ 7 

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

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

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 

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

3.2.2 Normality ................................................................................................................... 11 

3.2.3 Transformation of data ............................................................................................... 11

3.2.4 Heterogeneous variances ............................................................................................ 11 

3.2.5 Monotonicity test ........................................................................................................ 12 

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

4. Results ................................................................................................................................................. 12 

Tables

Table 0. Data information for SMV Octylphenol .................................................................................... 4 

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

Generation: F01,2,3 .................................................................................................................... 13 

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

Generation: F11,2,3 .................................................................................................................... 14 

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

Octylphenol, Generation: F0 ................................................................................................... 15 

Table 1D. Statistical Analysis Results for Reproduction Endpoints. Laboratory: SMV, Test Chemical: 

Octylphenol, Generation: F1 ................................................................................................... 16 
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Table 1E. Comparisons between Two Generations for Reproduction Endpoints. Laboratory: SMV, 

Test Chemical: Octylphenol. ................................................................................................... 17 

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

F01,2,3,4 ...................................................................................................................................... 18 

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

Generation: F11,2,3 .................................................................................................................... 19 

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

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

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

Octylphenol, Generation: F1 ................................................................................................... 21 

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

Chemical: Octylphenol. ........................................................................................................... 22 

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

Generation: F01,2,3 .................................................................................................................... 23 

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

Generation: F11,2,3 .................................................................................................................... 24 

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

Octylphenol, Generation: F0 ................................................................................................... 25 

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

Octylphenol, Generation: F1 ................................................................................................... 26 

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

Chemical: Octylphenol. ........................................................................................................... 27 

Figures

Figure 1. Means ± Standard Errors of Untransformed data: Time to Mature By Generation and Group 

For Chemical Octylphenol From Lab SMV. ........................................................................... 28 

Figure 2. Means ± Standard Errors of Untransformed data: Number Offspring (brood 1) By Generation 

and Group For Chemical Octylphenol From Lab SMV. ......................................................... 28 

Figure 3. Means ± Standard Errors of Untransformed data: Number Offspring (brood 2) By Generation 

and Group For Chemical Octylphenol From Lab SMV. ......................................................... 29 

Figure 4. Means ± Standard Errors of Untransformed data: Number Offspring (two broods) by 

Generation and Group For Chemical Octylphenol From Lab SMV. ...................................... 29 

Figure 5. Means ± Standard Errors of Untransformed data: Days to Brood 1 By Generation and Group 

For Chemical Octylphenol From Lab SMV. ........................................................................... 30 

Figure 6. Means ± Standard Errors of Untransformed data: Days to Brood 2 By Generation and Group 

For Chemical Octylphenol From Lab SMV. ........................................................................... 30 
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Figure 7. Means ± Standard Errors of Untransformed data: Days Between Broods 1 and 2 By 

Generation and Group For Chemical Octylphenol From Lab SMV. ...................................... 31 

Figure 8. Means ± Standard Errors of Untransformed data: Total Productive Days By Generation and 

Group For Chemical Octylphenol From Lab SMV. ................................................................ 31 

Figure 9. Means ± Standard Errors of Untransformed data: Offspring per Female per Productive Day 

By Generation and Group For Chemical Octylphenol From Lab SMV. ................................. 32 

Figure 10. Means ± Standard Errors of Untransformed data: Percent of Females Productively Active By 

Generation and Group For Chemical Octylphenol From Lab SMV. ...................................... 32 

Figure 11. Means ± Standard Errors of Untransformed data: Sex Ratio By Generation and Group For 

Chemical Octylphenol From Lab SMV. ................................................................................. 33 

Figure 12. Dry Weight (mg) (day 0) By Generation and Group For Chemical Octylphenol From Lab 

SMV. ....................................................................................................................................... 33 

Figure 13. Means ± Standard Errors of Untransformed data: Dry Weight (mg) (day 7) By Generation 

and Group For Chemical Octylphenol From Lab SMV. ......................................................... 34 

Figure 14. Means ± Standard Errors of Untransformed data: Dry Weight (mg) (day 14) By Generation 

and Group For Chemical Octylphenol From Lab SMV. ......................................................... 34 

Figure 15. Means ± Standard Errors of Untransformed data: Dry Weight (mg) (Termination) By 

Generation and Group For Chemical Octylphenol From Lab SMV. ...................................... 35 

Figure 16. Means ± Standard Errors of Untransformed data: Length (mm) (day 0) By Generation and 

Group For Chemical Octylphenol From Lab SMV. ................................................................ 35 

Figure 17. Means ± Standard Errors of Untransformed data: Length (mm) (day 7) By Generation and 

Group For Chemical Octylphenol From Lab SMV. ................................................................ 36 

Figure 18. Means ± Standard Errors of Untransformed data: Length (mm) (day 14) By Generation and 

Group For Chemical Octylphenol From Lab SMV. ................................................................ 36 

Figure 19. Means ± Standard Errors of Untransformed data: Length (mm) (Termination) By Generation 

and Group For Chemical Octylphenol From Lab SMV. ......................................................... 37 

Figure 20. Means ± Standard Errors of Untransformed data: Reproduction Survival Rate (%) (day 7) By 

Generation and Group For Chemical Octylphenol From Lab SMV. ...................................... 37 

Figure 21. Means ± Standard Errors of Untransformed data: Reproduction Survival Rate (%) (day 14) 

By Generation and Group For Chemical Octylphenol From Lab SMV. ................................. 38 

Figure 22. Means ± Standard Errors of Untransformed data: Reproduction Survival Rate (%) 

(Termination) By Generation and Group For Chemical Octylphenol From Lab SMV. ......... 38 

Figure 23. Means ± Standard Errors of Untransformed data: Total Survival Rate (%) (day 7) By 

Generation and Group For Chemical Octylphenol From Lab SMV. ...................................... 39 
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Figure 24. Means ± Standard Errors of Untransformed data: Total Survival Rate (%) (day 14) By 

Generation and Group For Chemical Octylphenol From Lab SMV. ...................................... 39 

Figure 25. Means ± Standard Errors of Untransformed data: Total Survival Rate (%) (Termination) By 

Generation and Group For Chemical Octylphenol From Lab SMV. ...................................... 40 
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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. SMV and WLI conducted two-generation mysid survival and growth tests following a similar 

protocol with the test chemical octylphenol. Statistical analyses are being carried out on the SMV and 

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

octylphenol test data. 

 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 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. 

 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 30 for SMV 

octylphenol), mysids from the reproduction baskets are also measured for weights and lengths. 
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 Intra-laboratory statistical analysis of the SMV octylphenol mysid study was performed 

according to a common statistical analysis plan. For this intra-laboratory analysis comparisons were 

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

mysid responses to graded concentrations of octylphenol. 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 specifies 25 endpoints. Some of the endpoints are recorded per mysid or 

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

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

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

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

basket (i.e. per replicate tank). These endpoints are 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) 

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

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

survival, per basket) 

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

7, 14 and termination (total survival, per basket) 

 

 The survival rates were adjusted for the number removed for weight and length determinations 

(i.e. were omitted from both the numerator and denominator) and the other non-treatment related deaths 

(iii) Growth (F0, F1) (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) 

 

1

 Day 14 and terminal weights and lengths were reported by gender. 
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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. 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. 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 mysids on day 14), and for the surviving 

reproduction animals at termination. For the weight data, there are a total of 9 composites (5 mysids per 

composite) for the F0 generation and a total of 54 composites (9 composites (5 mysids per composite) per 

concentration group among the six concentration groups) for the F1 generation on day 0. On day 7, there 

are 3 weight composites (2 mysids per composite) per replicate and three replicates per concentration 

group. 

 The test 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. 

 
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Table 0. Data information for SMV Octylphenol 

Data Files and data sheets Endpoints Comments 

Original- SMV octylpehnol 

data.xls:

- F0 Reproduction 

- F1 Reproduction 

- F0 Summary 

- F1 Summary 

Reformatted-: 

 SMV Octylphenol fecunity.xls: 

- F0Reproduction 

- F1Reproduction; 

 SMV Octylphenol 

SexRatio.xls: 

- F0SexRatio 

- F1SexRatio 

 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 

 Offspring per female per 

reproductive day for the 

first two broods 

 Total reproductively 

active days for the first 

two broods 

 Sex ratio 

 If there are no offspring for 

a brood. The data are 

recorded as “NA” for the 

brood sizes and time to 

brood. Per EPA suggestion, 

such data are changed as: 

zero if female is still living, 

and “NA” for a brood (1st or 

2nd brood) if female died 

without releasing the brood. 

 The total offspring for the 

first 2 broods and the # of 

offspring per female per 

reproductive day are 

therefore recalculated and 

displayed in the reformatted 

data. 

 For each brood pair, the 

total reproductive days for 

the first two broods is the 

days from the first brood 

from any female to the days 

of second broods of the 

brood pairs. If the broods 

have a “NA” or zero sizes, 

the day for the brood is a 

“NA”, and total 

reproductive days for the 

SMV lindane data was 

recorded as “NA”. The 

SMV octylphenol used the 

date until the female died, 

which is inconsistent from 

the SMV lindane. Thus, we 

treated them as “NA” or 

missing just like the SMV 

lindane data. 

 Time to maturity is given 

once for each replicate tank. 

 %of female reproductively 

active, sex ratio are reported 

per tank. The other 

reproductive endpoints are 

reported per mating pair. 

Original- SMV octylpehnol data.xls:

One sheet for each generation and 

Weight (dry weight) and 

length for days 0, 7, 14 and 

 For F0 generation all 

mysids on day 0 are kept in 
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each day 0, 7, 14 and termination 

Reformatted-SMV Octylphenol 

Growth.xls 

One sheet for each generation and 

each day 0, 7, 14 and termination 

termination untreated water. There 

were 9 weight composites 

(5 mysids per composite). 

 For F1 day 0 9 composites 

(5 mysids per composite) 

for each control and 

concentration group. 

 Length is reported per 

mysid. 

 Weight is reported per 5 

mysid composites for day 0, 

per 2 mysid composites for 

day 7, and per mysid for 

day 14 and for termination. 

 Gender of mysid is 

provided for day 14 and 

termination data. 

Original- SMV octylpehnol 

data.xls:

- F0 Reproduction Survival 

- F0 Growth Survival 

- F0 Total Survival 

- F1 Reproduction Survival 

- F1 Growth Survival 

- F1 Total Survival 

Reformatted- SMV Octylphenol 

Survival.xls:

- F0ReproductionSurvival; 

- F0GrowthSurvival 

- F0TotalSurvival 

- F1ReproductionSurvival 

- F1GrowthSurvival 

- F1TotalSurvival 

 Survival (reproduction) 

(day 7, day 14 and 

termination) 

 Survival (total) (day 7, 

day 14 and termination) 

 Survival endpoints are 

adjusted for the nontreatment related death, 

 The Total survival at day 27 

endpoint is incorrect 

because the total survival is 

the sum of the reproductive 

survival at day 27 and the 

growth survival at day 14. 

Growth survival is not 

carried beyond day 14, at 

least in the SMV 

Octylphenol data set. Thus 

the total survival at day 27 

assumes that there is no 

lethality in the growth bin 

between day 15 and day 27 

and so overestimates the 

survival. 

 Survival endpoints are 

reported per-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 available only with the one-way ANOVA in 

STATCHARRMS. Thus for the analyses within generations it was elected to base analyses on the 

one-way ANOVA procedure. In the one way ANOVA STATCHARRMS pools the results across 
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replicate tanks (and individual mysids (if the data were presented for each individual)) within test 

or control groups. The one way ANOVA (parametric or nonparametric forms) is 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 ANOVA model, with 

replicate tanks within treatment groups treated as random effects. The objectives of the analyses 

differ from those of the within generation analysis. The analyses within generations compare the 

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

results between generations, 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. In STATCHARRMS, there is 

no test for monotonicity within the mixed effects models (thus, the two-way ANOVA 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 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 analysis (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 called 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 

were enumerated in the STATCHARRMS outputs, and will be sent to EPA for review on a later 

date. 

 Total survival at termination is an upwardly biased value in the SMV octylphenol data. This is 

because survival for the growth mysids is recorded only through day 14. The total survival at 

termination therefore tacitly assumes that there is no mortality in the growth mysids between day 

14 and termination (about study day 30). This is clearly an underestimate of growth mysid 

mortality beyond day 14. 
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 For the reproductive response when “NA” 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 “NA” brood size is generated, the brood size should be counted as 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 entry for a given 

brood size is zero or “NA”, the time for the brood was also recorded as “NA” in the original data 

file and 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. If the brood size is missing or zero in 

either brood 1 or brood 2, the number of reproductive days is recorded as “NA” and the number 

of offspring per female per reproductive day is treated as missing. 

 In the SMV 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. 

3. STATISTICAL METHODS 

Statistical analysis procedures used conform to the analysis procedures 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 or per mysid-pair data without first averaging across the mysids in 

the same replicate basket. 

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. 
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3.1.1 Outlier Screens 

 For each generation, 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 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 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 contrast 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. 

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. 
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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. 

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. Residual variance was treated as homogeneous 

or heterogeneous between generations based 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 here are 

based on the default auto options in STATCHARRMS. 
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11 

3.2.1 Outlier screens 

 In STATCHARMS, the outliers screened for the comparisons of two generations were based on 

the two-way mixed effect model in section 3.2.6. Outliers were detected based on the studentized 

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 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). Outliers were included in 

the test. 
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3.2.5 Monotonicity test 

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

the STATCHARRMS two-way mixed effects ANOVA. 

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 tested to identify the concentration groups for which the two generations were 

significantly different. 

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 25. 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 (reproduction), Tables 2C and 2D (growth), and Tables 3C and 3D (survival). 

Comparisons between the F0 and F1 generations are summarized in Tables 1E (reproduction), 2E 

(growth), and 3E (survival). 

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

Concentration Statistics Time to Mature 

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 

Total 

Reproductive 

Days 

for the First 

Two Broods 

# of Offspring 

Per Female 

Per 

Reproduction 

Day 

 for the First 

Two Broods 

P

Control 

(Group 1) 

Mean 13.000 6.048 8.238 14.286 19.000 23.500 4.500 9.500 1.636 

Standard 

Dev. 0.000 2.519 3.700 5.021 1.487 1.821 0.688 1.821 0.499 

N 3 21 21 21 20 20 20 20 20 

CV (%) 0 41.66 44.91 35.15 7.83 7.75 15.29 19.17 30.46 

0.26 μg/L 

 (Group 2) 

Mean 13.000 5.050 5.889 10.944 17.500 23.778 6.278 9.778 1.126 

Standard 

Dev. 0.000 1.669 4.213 4.569 1.147 2.777 2.675 2.777 0.354 

N 3 20 18 18 20 18 18 18 18 

CV(%) 0 33.06 71.55 41.75 6.55 11.68 42.6 28.4 31.46 

0.64 μg/L 

 (Group 3) 

Mean 13.000 4.450 6.389 10.944 18.450 23.053 4.579 9.053 1.242 

Standard 

Dev. 0.000 1.820 3.806 4.783 2.350 2.345 0.961 2.345 0.347 

N 3 20 18 18 20 19 19 19 18 

CV(%) 0 40.9 59.57 43.7 12.74 10.17 20.99 25.9 27.91 

1.6 μg/L 

 (Group 4) 

Mean 13.000 4.524 6.842 11.579 19.238 24.278 5.222 10.278 1.194 

Standard 

Dev. 0.000 1.601 3.219 3.322 2.755 2.421 1.060 2.421 0.269 

N 3 21 19 19 21 18 18 18 18 

CV(%) 0 35.38 47.05 28.69 14.32 9.97 20.3 23.55 22.54 

4.0 μg/L 

 (Group 5) 

Mean 13.000 6.000 5.238 11.238 18.143 24.000 6.050 10.000 1.229 

Standard 

Dev. 0.000 2.757 2.548 4.098 2.308 3.061 2.328 3.061 0.423 

N 3 21 21 21 21 20 20 20 20 

CV(%) 0 45.95 48.64 36.46 12.72 12.75 38.48 30.61 34.37 

10 μg/L 

 (Group 6) 

Mean 13.000 4.842 6.111 11.056 19.895 25.500 5.500 11.500 0.975 

Standard 

Dev. 0.000 1.463 3.306 4.165 2.747 2.407 1.886 2.407 0.345 

N 3 19 18 18 19 18 18 18 18 

CV(%) 0 30.21 54.09 37.68 13.81 9.44 34.3 20.93 35.36 

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

Concentration Statistics 

Time to 

Mature 

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 

Total 

Reproductive 

Days 

for the First Two 

Broods 

# of Offsprin

Per Female Pe

Reproduction 

Day 

 for the First 

Two Broods 

Control 

(Group 1) 

Mean 10.667 6.313 7.154 13.077 16.333 21.182 5.364 9.182 1.754 

Standard 

Dev. 0.577 2.869 4.451 6.563 2.717 2.523 1.859 2.523 0.664 

N 3 16 13 13 15 11 11 11 11 

CV(%) 5.41 45.44 62.21 50.19 16.63 11.91 34.65 27.47 37.86 

0.26 μg/L 

 (Group 2) 

Mean 12.000 7.842 11.000 19.111 17.789 22.889 4.944 10.889 1.787 

Standard 

Dev. 0.000 3.819 4.058 6.902 2.658 2.742 1.110 2.742 0.545 

N 3 19 18 18 19 18 18 18 18 

CV(%) 0 48.7 36.89 36.11 14.94 11.98 22.45 25.18 30.49 

0.64 μg/L 

 (Group 3) 

Mean 12.333 6.056 7.800 13.533 15.778 20.929 5.286 8.929 1.618 

Standard 

Dev. 1.528 2.209 3.406 3.962 0.943 1.385 1.139 1.385 0.459 

N 3 18 15 15 18 14 14 14 14 

CV(%) 12.39 36.48 43.67 29.27 5.98 6.62 21.54 15.51 28.35 

1.6 μg/L 

 (Group 4) 

Mean 11.667 5.706 7.500 13.125 16.765 21.333 5.000 9.333 1.669 

Standard 

Dev. 0.577 1.795 4.775 4.965 2.562 1.073 1.348 1.073 0.285 

N 3 17 16 16 17 12 12 12 12 

CV(%) 4.95 31.45 63.67 37.83 15.28 5.03 26.97 11.5 17.09 

4.0 μg/L 

 (Group 5) 

Mean 13.000 6.450 9.500 15.722 16.750 21.353 4.882 9.353 1.741 

Standard 

Dev. 1.732 2.781 3.312 4.860 1.916 1.412 0.600 1.412 0.357 

N 3 20 18 18 20 17 17 17 17 

CV(%) 13.32 43.12 34.87 30.91 11.44 6.61 12.29 15.09 20.48 

10 μg/L 

 (Group 6) 

Mean 11.000 4.688 6.200 10.933 16.250 20.231 4.385 8.231 1.471 

Standard 

Dev. 1.000 1.922 4.109 4.317 2.082 2.279 0.506 2.279 0.568 

N 3 16 15 15 16 13 13 13 13 

CV(%) 9.09 41.01 66.28 39.49 12.81 11.26 11.55 27.68 38.62 

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 measured per mating pair. 

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: SMV, Test 

 Chemical: Octylphenol, Generation: F0 

Parameters 

Normality 

Test1

Homogeneity 

of Variance 

Test2

Monotonicity 

Test3

Concentration 

Trend Tests4

Overall 

Concentration 

Effect5

Pairwise 

Comparisons to 

Control6,7

Time to Mature8 Non-Normal NA NonMonotonic NA No NA 

Number 

Offspring 

(Brood 1)

Non-Normal NA NonMonotonic NA Yes Groups 3 & 4 

Number 

Offspring 

(Brood 2)

Normal9

 Homogeneous NonMonotonic NA Yes Group 5 

Number 

Offspring 

(two Broods)

Normal Homogeneous NonMonotonic NA Yes Groups 2 and 3, 

5 and 6 

Days to 

Brood 1 Non-Normal NA NonMonotonic NA Yes Group 2 

Days to 

Brood 2 Normal10 Homogeneous NonMonotonic NA No NA 

Days Between Broods 

1 and 2 Non-Normal NA NonMonotonic NA Yes Groups 2 & 5 

Total Reproductive 

Days for the First Two 

Broods 

Non-Normal NA NonMonotonic NA No NA 

# of Offspring Per 

Female Per 

Reproduction Day 

 for the First 

Two Broods 

Normal Homogeneous Monotonic 

Decreasing trend for 

all concentration 

groups 

Yes NA 

% of Females 

Productively 

Active 

Non-Normal NA NonMonotonic NA No NA 

Sex Ratio Normal Homogeneous 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 

within-treatment variances for normal distributed data 

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

8. All responses are constant. 

9. Normal and homogeneous when square root-transformed 

10. Normal and homogeneous when log-transformed. 
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Table 1D. Statistical Analysis Results for Reproduction Endpoints. Laboratory: SMV, Test 

 Chemical: Octylphenol, Generation: F1 

Parameters 

Normality 

Test1

Homogeneity 

of Variance 

Test2

Monotonicity 

Test3

Concentration Trend 

Tests4

Overall 

Concentra

tion Effect5

Pairwise 

Comparisons to 

Control6,7

Time to Mature Normal Homogeneous NonMonotonic NA No NA 

Number Offspring 

(Brood 1) Normal Homogeneous Monotonic Decreasing trends for 

the highest dose group Yes NA 

Number Offspring 

(Brood 2)

NonNormal NA NonMonotonic NA Yes No 

Number Offspring 

(two Broods) Normal Homogeneous NonMonotonic NA Yes Group 2 

Days to Brood 1 Normal Homogeneous NonMonotonic NA No NA 

Days to Brood 2 NonNormal NA NonMonotonic NA No NA 

Days Between Broods 1 

and 2 

NonNormal NA NonMonotonic NA No NA 

Total Reproductive Days 

for the First Two Broods 

NonNormal NA NonMonotonic NA No NA 

# of Offspring Per Female 

Per Reproduction Day 

 for the First 

Two Broods 

Normal Homogeneous NonMonotonic NA No NA 

% of Females 

Reproductively Active 

NonNormal NA 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. 

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

 SMV, Test Chemical: Octylphenol. 

Parameters 

Normality 

Test1

Homogeneity 

of variance 

Test2

Parameteric/ 

Nonparametric 

Concentration 

Main Effect 

Generation Main 

Effect 

Concentration 

by Generation 

Interaction 

Groups with 

Significant 

Difference 

between 

Generations3

Time to 

Mature4 NA NA NA NA NA NA NA 

Number 

Offspring 

(Brood 1)

Normal Homogeneous Parametric Yes Yes No Group 2 

Number 

Offspring 

(Brood 2)

Normal Homogeneous Parametric No Yes Yes Groups 2 and 

5 

Number 

Offspring 

(two Broods)

Normal Homogeneous Parametric Yes Yes Yes Groups 2 and 

5 

Days to 

Brood 1 Normal Homogeneous Parametric No Yes No Groups 1, 3, 

4, and 6 

Days to 

Brood 2 Normal Homogeneous Parametric No Yes No Groups 1, 4, 

5 and 6 

Days Between 

Broods 1 and 

2 

Normal Homogeneous Parametric No No No NA 

Total 

Reproductive 

Days for the 

First Two 

Broods 

Normal Homogeneous Parametric No No No NA 

# of Offspring 

Per Female Per 

Reproduction 

Day 

 for the First 

Two Broods 

Normal Homogeneous Parametric Yes Yes No Groups 2, 4, 

5, 6 

% of Females 

Productively 

Active 

NonNormal NA Nonparametric No No No NA 

Sex Ratio Normal Homogeneous Parametric No Yes No Groups 2 and 

3 

1. Shapiro-Wilk test for normality. 

2. Levene test for homogeneity of variances. 

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

4. Model is not fit due to constant value for time to mature in F0 generation. 

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

Generation: F01,2,3,4

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) 

Lengt

h 

(mm) 

(day 

7) 

Length 

(mm) 

(day 14) 

Length 

(mm) 

(Termination

) 

Control 

(Group 1) 

Mean 0.034 0.083 0.440 0.994 1.703 3.469 5.321 7.467 

Standard 

Dev. 0.005 0.036 0.139 0.205 0.165 0.361 0.379 0.306 

N 9 9 36 41 45 18 36 41 

CV(%) 13.86 42.64 31.54 20.59 9.66 10.4 7.12 4.1 

0.26 μg/L 

 (Group 2) 

Mean 0.144 0.544 1.032 4.062 5.763 7.412 

Standard 

Dev. 0.042 0.093 0.250 0.276 0.322 0.355 

N 9 32 31 18 32 31 

CV(%) 28.88 17.16 24.25 6.8 5.59 4.79 

0.64 μg/L 

 (Group 3) 

Mean 0.124 0.456 1.027 3.926 5.699 7.375 

Standard 

Dev. 0.057 0.128 0.214 0.307 0.241 0.351 

N 9 32 41 18 32 41 

CV(%) 45.61 27.95 20.8 7.81 4.22 4.75 

1.6 μg/L 

 (Group 4) 

Mean 0.137 0.440 1.011 3.893 5.649 7.240 

Standard 

Dev. 0.082 0.096 0.211 0.320 0.300 0.318 

N 9 38 38 18 38 38 

CV(%) 59.72 21.84 20.83 8.23 5.32 4.39 

4.0 μg/L 

 (Group 5) 

Mean 0.159 0.440 1.012 3.809 5.558 7.313 

Standard 

Dev. 0.077 0.078 0.207 0.429 0.519 0.476 

N 9 37 42 18 37 42 

CV(%) 48.49 17.81 20.42 11.27 9.33 6.52 

10 μg/L 

 (Group 6) 

Mean 0.146 0.553 0.995 3.927 5.848 7.128 

Standard 

Dev. 0.047 0.086 0.203 0.323 0.270 0.300 

N 9 33 32 18 33 32 

CV(%) 31.97 15.49 20.44 8.24 4.62 4.21 

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

 Generation: F11,2,3

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 0.023 0.252 0.560 1.066 1.607 4.446 6.057 7.311 

Standard 

Dev. 0.004 0.036 0.131 0.274 0.173 0.394 0.367 0.384 

N 9 9 19 29 45 18 19 29 

CV(%) 15.91 14.26 23.34 25.74 10.76 8.86 6.06 5.25 

0.26 μg/L 

 (Group 2) 

Mean 0.020 0.292 0.553 1.039 1.581 4.943 6.554 7.542 

Standard 

Dev. 0.004 0.045 0.144 0.216 0.121 0.423 0.686 0.452 

N 9 9 22 29 45 18 22 29 

CV(%) 22.16 15.38 26 20.79 7.66 8.55 10.47 5.99 

0.64 μg/L 

 (Group 3) 

Mean 0.020 0.263 0.597 1.060 1.586 4.466 6.212 7.479 

Standard 

Dev. 0.004 0.072 0.139 0.220 0.111 0.421 0.353 0.397 

N 9 9 19 27 45 18 19 27 

CV(%) 22.74 27.32 23.24 20.77 6.99 9.43 5.68 5.31 

1.6 μg/L 

 (Group 4) 

Mean 0.020 0.252 0.579 0.996 1.610 4.739 6.334 7.296 

Standard 

Dev. 0.003 0.041 0.146 0.251 0.145 0.527 0.396 0.381 

N 9 9 16 29 45 18 16 29 

CV(%) 16.58 16.08 25.15 25.18 8.99 11.11 6.25 5.23 

4.0 μg/L 

 (Group 5) 

Mean 0.026 0.230 0.562 1.014 1.608 4.863 6.823 7.399 

Standard 

Dev. 0.008 0.042 0.113 0.260 0.146 0.350 0.400 0.575 

N 9 9 21 35 45 18 21 35 

CV(%) 30.25 18.19 20.15 25.66 9.08 7.21 5.86 7.77 

10 μg/L 

 (Group 6) 

Mean 0.021 0.276 0.627 0.989 1.582 4.651 6.412 7.099 

Standard 

Dev. 0.011 0.040 0.144 0.281 0.130 0.328 0.520 0.368 

N 9 9 21 31 45 18 21 31 

CV(%) 55.59 14.53 23 28.43 8.23 7.05 8.12 5.18 

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

2. Length is measured per mysid. Weights were measured on averages per composite 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. 

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

 Chemical: Octylphenol, Generation: F0 

Parameters 

Normality 

Test1

Homogeneity of 

variance Test2

Monotonicity 

Test3

Concentration 

Trend Tests4

Overall 

Concentration 

Effect5

Pairwise 

Comparisons to 

Control6,7

Dry Weight(mg) 

 (day 7) Non-Normal NA Monotonic 

Increasing trend for 

the two highest 

concentration groups 

No NA 

Dry Weight(mg) 

(day 14) Normal Heterogeneous Non-Monotonic NA Yes Groups 2 & 6 

Dry Weight (mg) 

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

Length (mm) 

(day 7) Normal Homogeneous Non-Monotonic NA Yes Groups 2, 3, 4, 5 

& 6 

Length (mm) 

(day 14) Non-Normal NA Monotonic 

Increasing trends for 

all concentration 

groups 

Yes NA 

Length (mm) 

(Termination) Non-Normal NA Monotonic 

Decreasing trends for 

the three highest 

concentration groups 

Yes 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. 

 
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Octylphenol Data 

21 

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

 Chemical: Octylphenol, Generation: F1 

Parameters 

Normality 

Test1

Homogeneity of 

variance Test2

Monotonicity 

Test3

Concentration 

Trend Tests4

Overall 

Concentration 

Effect5

Pairwise 

Comparisons to 

Control6,7

Dry Weight (mg) 

(day 0) Non-Normal NA Non-Monotonic NA No NA 

Dry Weight (mg) 

(day 7) Normal Homogeneous Non-Monotonic NA No NA 

Dry Weight (mg) 

(day 14) Non-Normal NA Non-Monotonic NA No NA 

Dry Weight (mg) 

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

Length (mm) 

(day 0) Normal Homogeneous Non-Monotonic No No NA 

Length (mm) 

(day 7) Non-Normal NA Non-Monotonic NA Yes Groups 2 & 5 

Length (mm) 

(day 14) Normal Heterogeneous Monotonic 

Increasing dose 

trends for the two 

highest 

concentration 

groups. 

Yes NA 

Length (mm) 

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

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. 

 
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Octylphenol Data 

22 

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

 Test Chemical: Octylphenol. 

Parameters 

Normality 

Test1

Homogeneity 

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 7) Normal Homogeneous Parametric No Yes No Groups 1, 2, 

3, 4 and 6 

Dry Weight 

(mg) 

(day 14)

Normal Homogeneous Parametric No Yes No Groups 3, 4 

and 5 

Dry Weight 

(mg) 

(Termination)

Normal Homogeneous Parametric No No No NA 

Length (mm) 

(day 7) Normal Homogeneous Parametric Yes Yes No Groups 1, 2, 

3, 4, 5 and 6 

Length (mm) 

(day 14) Normal Homogeneous Parametric Yes No No Groups 1, 2, 

3, 4, 5 and 6 

Length (mm) 

(Termination) Normal Homogeneous Parametric No No No NA 

1. Shapiro-Wilk test for normality. 

2. Levene test for homogeneity of variances. 

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

 
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Octylphenol Data 

23 

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

 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) 

Total 

Survival Rate (%) 

(Termination) 

Control 

(Group 1) 

Mean 91.667 86.667 73.000 94.333 86.000 79.000 

Standard 

Dev. 10.408 2.887 8.660 4.041 6.245 9.000 

N 3 3 3 3 3 3 

CV(%) 11.35 3.33 11.86 4.28 7.26 11.39 

0.26 μg/L 

 (Group 2) 

Mean 93.333 91.667 64.333 95.000 86.000 70.333 

Standard 

Dev. 5.774 5.774 12.662 5.000 3.464 5.508 

N 3 3 3 3 3 3 

CV(%) 6.19 6.3 19.68 5.26 4.03 7.83 

0.64 μg/L 

 (Group 3) 

Mean 93.333 91.667 78.000 95.333 85.000 77.000 

Standard 

Dev. 7.638 5.774 10.000 4.619 7.937 8.718 

N 3 3 3 3 3 3 

CV(%) 8.18 6.3 12.82 4.84 9.34 11.32 

1.6 μg/L 

 (Group 4) 

Mean 91.667 91.667 70.333 94.333 91.000 79.333 

Standard 

Dev. 7.638 7.638 6.429 5.132 6.000 3.512 

N 3 3 3 3 3 3 

CV(%) 8.33 8.33 9.14 5.44 6.59 4.43 

4.0 μg/L 

 (Group 5) 

Mean 91.667 83.333 75.333 92.667 85.333 81.000 

Standard 

Dev. 10.408 15.275 13.051 7.506 13.317 12.124 

N 3 3 3 3 3 3 

CV(%) 11.35 18.33 17.32 8.1 15.61 14.97 

10 μg/L 

 (Group 6) 

Mean 91.667 86.667 65.000 92.000 83.000 71.667 

Standard 

Dev. 5.774 2.887 9.539 1.732 1.732 8.145 

N 3 3 3 3 3 3 

CV(%) 6.3 3.33 14.68 1.88 2.09 11.36 

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 

Intra-Laboratory Analysis Report for Mysid: SMV Octylphenol Data 

24 

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

 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) 

Total 

Survival Rate (%) 

(Termination) 

Control 

(Group 1) 

Mean 95.333 89.333 77.000 92.000 86.000 78.333 

Standard 

Dev. 4.041 10.066 16.093 1.732 5.196 6.658 

N 3 3 3 3 3 3 

CV(%) 4.24 11.27 20.9 1.88 6.04 8.5 

0.26 μg/L 

 (Group 2) 

Mean 95.333 95.333 90.000 94.667 90.333 84.000 

Standard 

Dev. 4.041 4.041 8.660 6.807 6.658 8.185 

N 3 3 3 3 3 3 

CV(%) 4.24 4.24 9.62 7.19 7.37 9.74 

0.64 μg/L 

 (Group 3) 

Mean 84.333 83.000 75.000 87.667 81.333 76.333 

Standard 

Dev. 7.506 3.606 7.211 5.033 2.887 7.234 

N 3 3 3 3 3 3 

CV(%) 8.9 4.34 9.61 5.74 3.55 9.48 

1.6 μg/L 

 (Group 4) 

Mean 88.667 81.667 68.333 87.667 77.667 69.333 

Standard 

Dev. 7.506 5.508 12.220 6.807 9.238 7.767 

N 3 3 3 3 3 3 

CV(%) 8.46 6.74 17.88 7.76 11.89 11.2 

4.0 μg/L 

 (Group 5) 

Mean 95.333 93.000 88.000 95.333 90.000 86.333 

Standard 

Dev. 4.041 0.000 3.606 4.041 1.732 0.577 

N 3 3 3 3 3 3 

CV(%) 4.24 0 4.1 4.24 1.92 0.67 

10 μg/L 

 (Group 6) 

Mean 100.000 97.667 93.000 94.333 90.333 86.667 

Standard 

Dev. 0.000 4.041 6.083 5.132 8.737 8.505 

N 3 3 3 3 3 3 

CV(%) 0 4.14 6.54 5.44 9.67 9.81 

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 

Intra-Laboratory Analysis Report for Mysid: SMV Octylphenol Data 

25 

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

 Chemical: Octylphenol, Generation: F0 

Parameters Normality 

Test1

Homogeneity of 

variance Test2

Monotonicity 

Test3

Concentration 

Trend Tests4

Overall 

Concentration 

Effect5

Pairwise 

Comparisons to 

Control6,7

Reproduction 

Survival Rate (%) 

(day 7)

Normal Homogeneous Non-Monotonic NA No NA 

Reproduction 

Survival Rate (%) 

(day 14)

Normal Homogeneous Non-Monotonic NA No NA 

Reproduction 

Survival Rate (%) 

(Termination)

Normal Homogeneous Non-Monotonic NA No NA 

Total 

Survival Rate (%) 

(day 7)

Normal Homogeneous Non-Monotonic NA No NA 

Total 

Survival Rate 

(%) (day 14)

Normal Homogeneous Non-Monotonic NA No NA 

Total 

Survival Rate (%) 

(Termination)

Normal Homogeneous 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 

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: SMV Octylphenol Data 

26 

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

 Chemical: Octylphenol, Generation: F1 

Parameters 

Normality 

Test1

Homogeneity of 

variance Test2

Monotonicity 

Test3

Concentration 

Trend Tests4

Overall 

Concentration 

Effect5

Significant 

Pairwise 

Comparisons 

to Control6,7

Reproduction 

Survival Rate (%) 

(day 7)

Normal Homogeneous Non-Monotonic NA Yes No 

Reproduction 

Survival Rate (%) 

(day 14)

Normal Homogeneous Non-Monotonic NA Yes No 

Reproduction 

Survival Rate (%) 

(Termination)

Normal Homogeneous Monotonic NA No NA 

Total 

Survival Rate (%) 

(day 7)

Normal Homogeneous Non-Monotonic NA No NA 

Total 

Survival Rate 

(%) (day 14)

Normal Homogeneous Non-Monotonic NA No NA 

Total 

Survival Rate (%) 

(Termination)

Normal Homogeneous 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 

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: SMV Octylphenol Data 

27 

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

 Chemical: Octylphenol. 

Parameters 

Normality 

Test1

Homogeneity 

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)

Normal Homogeneous Parametric No No No NA 

Reproduction 

Survival Rate 

(%) 

(day 14)

Normal Homogeneous Parametric No No No NA 

Reproduction 

Survival Rate 

(%) 

(Termination)

Normal Homogeneous Parametric No Yes Yes Groups 2 and 

6 

Total 

Survival Rate 

(%) 

(day 7)

Normal Homogeneous Parametric No No No NA 

Total 

Survival Rate 

(%) (day 14)

Normal Homogeneous Parametric No No No NA 

Total 

Survival Rate 

(%) 

(Termination)

Normal Homogeneous Parametric No No No NA 

1. Shapiro-Wilk test for normality. 

2. Levene test for homogeneity of variances. 

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

Intra-Laboratory Analysis Report for Mysid: SMV Octylphenol Data 

28 

Figure 1. Means ± Standard Errors of Untransformed data: Time to Mature By Generation 

and Group For Chemical Octylphenol From Lab SMV. 

Figure 2. Means ± Standard Errors of Untransformed data: Number Offspring (brood 1) By 

Generation and Group For Chemical Octylphenol From Lab SMV. 

Time To Mature (Days)

10.0

10.5

11.0

11.5

12.0

12.5

13.0

13.5

14.0

14.5

15.0

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0 Generation F1

Number Offspring (brood 1)

3

4

5

6

7

8

9

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0 Generation F1
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Octylphenol Data 

29 

Figure 3. Means ± Standard Errors of Untransformed data: Number Offspring (brood 2) By 

Generation and Group For Chemical Octylphenol From Lab SMV. 

Figure 4. Means ± Standard Errors of Untransformed data: Number Offspring (two broods) 

by Generation and Group For Chemical Octylphenol From Lab SMV. 

Number Offspring (brood 2)

4

5

6

7

8

9

10

11

12

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0 Generation F1

Number Offspring (two broods)

8

10

12

14

16

18

20

22

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0 Generation F1
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Octylphenol Data 

30 

Figure 5. Means ± Standard Errors of Untransformed data: Days to Brood 1 By Generation 

and Group For Chemical Octylphenol From Lab SMV. 

Figure 6. Means ± Standard Errors of Untransformed data: Days to Brood 2 By Generation 

and Group For Chemical Octylphenol From Lab SMV. 

Days to Brood 1

15

16

17

18

19

20

21

22

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0 Generation F1

Days to Brood 2

19

20

21

22

23

24

25

26

27

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0 Generation F1
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Octylphenol Data 

31 

Figure 7. Means ± Standard Errors of Untransformed data: Days Between Broods 1 and 2 By 

Generation and Group For Chemical Octylphenol From Lab SMV. 

Figure 8. Means ± Standard Errors of Untransformed data: Total Productive Days By 

Generation and Group For Chemical Octylphenol From Lab SMV. 

Days Between Broods 1 and 2

4

5

6

7

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0 Generation F1

Total Reproductive Days

7

8

9

10

11

12

13

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0 Generation F1
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Octylphenol Data 

32 

Figure 9. Means ± Standard Errors of Untransformed data: Offspring per Female per 

Productive Day By Generation and Group For Chemical Octylphenol From Lab 

SMV. 

Figure 10. Means ± Standard Errors of Untransformed data: Percent of Females Productively 

Active By Generation and Group For Chemical Octylphenol From Lab SMV. 

Offspring per Female per Reproductive Day 0.5

1.0

1.5

2.0

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0 Generation F1

% of Females Productively Active

80

85

90

95

100

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0 Generation F1
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Octylphenol Data 

33 

Figure 11. Means ± Standard Errors of Untransformed data: Sex Ratio By Generation and 

Group For Chemical Octylphenol From Lab SMV. 

Figure 12. Dry Weight (mg) (day 0) By Generation and Group For Chemical Octylphenol 

From Lab SMV. 

Sex Ratio

0.0

0.5

1.0

1.5

2.0

2.5

3.0

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0 Generation F1

Dry Weight (mg)(day 0)

0.014

0.016

0.018

0.020

0.022

0.024

0.026

0.028

0.030

0.032

0.034

0.036

0.038

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0 Generation F1
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Octylphenol Data 

34 

Figure 13. Means ± Standard Errors of Untransformed data: Dry Weight (mg) (day 7) By 

Generation and Group For Chemical Octylphenol From Lab SMV. 

Figure 14. Means ± Standard Errors of Untransformed data: Dry Weight (mg) (day 14) By 

Generation and Group For Chemical Octylphenol From Lab SMV. 

Dry Weight (mg)(day 7)

0.06

0.08

0.10

0.12

0.14

0.16

0.18

0.20

0.22

0.24

0.26

0.28

0.30

0.32

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0 Generation F1

Dry Weight (mg)(day 14)

0.40

0.45

0.50

0.55

0.60

0.65

0.70

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0 Generation F1
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Octylphenol Data 

35 

Figure 15. Means ± Standard Errors of Untransformed data: Dry Weight (mg) (Termination) 

By Generation and Group For Chemical Octylphenol From Lab SMV. 

Figure 16. Means ± Standard Errors of Untransformed data: Length (mm) (day 0) By 

Generation and Group For Chemical Octylphenol From Lab SMV. 

Dry Weight (mg)(Termination)

0.90

0.95

1.00

1.05

1.10

1.15

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0 Generation F1

Length (mm)(day 0)

1.54

1.56

1.58

1.60

1.62

1.64

1.66

1.68

1.70

1.72

1.74

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0 Generation F1
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Octylphenol Data 

36 

Figure 17. Means ± Standard Errors of Untransformed data: Length (mm) (day 7) By 

Generation and Group For Chemical Octylphenol From Lab SMV. 

Figure 18. Means ± Standard Errors of Untransformed data: Length (mm) (day 14) By 

Generation and Group For Chemical Octylphenol From Lab SMV. 

Length (mm)(day 7)

3.0

3.5

4.0

4.5

5.0

5.5

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0 Generation F1

Length (mm)(day 14)

5.0

5.5

6.0

6.5

7.0

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0 Generation F1
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Octylphenol Data 

37 

Figure 19. Means ± Standard Errors of Untransformed data: Length (mm) (Termination) By 

Generation and Group For Chemical Octylphenol From Lab SMV. 

Figure 20. Means ± Standard Errors of Untransformed data: Reproduction Survival Rate (%) 

(day 7) By Generation and Group For Chemical Octylphenol From Lab SMV. 

Length (mm)(Termination)

7.0

7.1

7.2

7.3

7.4

7.5

7.6

7.7

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0 Generation F1

Reproduction Survival Rate (%)(day 7)

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

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0 Generation F1
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Octylphenol Data 

38 

Figure 21. Means ± Standard Errors of Untransformed data: Reproduction Survival Rate (%) 

(day 14) By Generation and Group For Chemical Octylphenol From Lab SMV. 

Figure 22. Means ± Standard Errors of Untransformed data: Reproduction Survival Rate (%) 

(Termination) By Generation and Group For Chemical Octylphenol From Lab 

SMV. 

Reproduction Survival Rate (%)(day 14)

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

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0 Generation F1

Reproduction Survival Rate (%)(Termination)

55

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

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0 Generation F1
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Octylphenol Data 

39 

Figure 23. Means ± Standard Errors of Untransformed data: Total Survival Rate (%) (day 7) 

By Generation and Group For Chemical Octylphenol From Lab SMV. 

Figure 24. Means ± Standard Errors of Untransformed data: Total Survival Rate (%) (day 14) 

By Generation and Group For Chemical Octylphenol From Lab SMV. 

Total Survival Rate (%)(day 7)

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

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0 Generation F1

Total Survival Rate (%)(day 14)

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

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0 Generation F1
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Octylphenol Data 

40 

Figure 25. Means ± Standard Errors of Untransformed data: Total Survival Rate (%) 

(Termination) By Generation and Group For Chemical Octylphenol From Lab 

SMV. 

Total Survival Rate (%)(Termination)

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

Control 0.26

ug/L

0.64

ug/L

1.6

ug/L

4

ug/L

10

ug/L

Group

Generation F0 Generation F1