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Intra-Laboratory Analysis Report for Mysid: SMV Lindane Data 

Endocrine Disruptor Screening Program 

Contract No: EP-W-11-063 

Task Order No: 2 

Statistical Support for Technical Directive #3 

Data Analysis Support for Invertebrate Tier 2 Test and Generation of 

Integrated Summary Report 

Intra-Laboratory Statistical Analysis for SMV Lindane 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 ........................................................................................................ 7 

3.1.2 Normality ................................................................................................................. 7 

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

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

3.1.5 Monotonicity of response ........................................................................................ 8 

3.1.6 Comparisons of test chemical to controls ................................................................ 8 

3.2 Comparisons of the F0 and F1 generations ................................................................ 10 

3.2.1 Outlier screens ....................................................................................................... 10 

3.2.2 Normality ............................................................................................................... 10 

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

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

3.2.5 Monotonicity Test ................................................................................................. 11 

3.2.6 F0 generation versus F1 generation ....................................................................... 11 

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

Tables

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

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

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

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

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

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

Chemical: Lindane, Generation: F0 ........................................................................... 15 

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

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

SMV, Test Chemical: Lindane. ................................................................................ 17 

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

Generation: F01,2,3,4 .................................................................................................... 18 

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

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

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

Chemical: Lindane, Generation: F0 ........................................................................... 20 

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

Chemical: Lindane, Generation: F1 ........................................................................... 21 

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

Test Chemical: Lindane. ............................................................................................ 22 

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

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

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

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

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

Chemical: Lindane, Generation: F0 ........................................................................... 25 

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

Chemical: Lindane, Generation: F1 ........................................................................... 26 

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

Chemical: Lindane. .................................................................................................... 27 

Figures

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

Group For Chemical Lindane From Lab SMV. ......................................................... 28 

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

Generation and Group For Chemical Lindane From Lab SMV. ............................... 28 

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

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

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

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

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

Group For Chemical Lindane From Lab SMV. ......................................................... 30 

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

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

Figure 8. Means ± Standard Errors of Untransformed data: Total Reproductive Days for the 

First Two Broods By Generation and Group For Chemical Lindane From Lab 

SMV. .......................................................................................................................... 31 

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

per Reproductive Day for the First Two Broods By Generation and Group For 

Chemical Lindane From Lab SMV. ........................................................................... 32 

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

Active By Generation and Group For Chemical Lindane From Lab SMV. .............. 32 

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

Group For Chemical Lindane From Lab SMV. ......................................................... 33 

Figure 12. Means ± Standard Errors of Untransformed data: Dry Weight (mg) (day 0) By 

Generation and Group For Chemical Lindane From Lab SMV. ............................... 33 

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

Generation and Group For Chemical Lindane From Lab SMV. ............................... 34 

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

Generation and Group For Chemical Lindane From Lab SMV. ............................... 34 

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

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

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

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

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

Generation and Group For Chemical Lindane From Lab SMV. ............................... 36 

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

Generation and Group For Chemical Lindane From Lab SMV. ............................... 36 

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

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

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

(day 7) By Generation and Group For Chemical Lindane From Lab SMV. ............. 37 

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

(day 14) By Generation and Group For Chemical Lindane From Lab SMV. ........... 38 

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

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

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

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

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

(Termination) By Generation and Group For Chemical Lindane 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. All three laboratories conducted two-generation mysid survival and growth tests following a 

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

UNT lindane study data. The draft analysis documented in this report is based on the SMV lindane test. 

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

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

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

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

concentrations of lindane 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 27 for SMV), mysids 

from the reproduction baskets are also measured for weights and lengths. 
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 Intra-laboratory statistical analysis of the SMV lindane 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 lindane. 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: 

(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 tank) 

 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) 

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

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

g. Weight at day 14 (per mysid) 

h. Weight at termination (per mysid). 

 

1

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

reproduction animals at termination. For the weight data, there are a total of 27 composites (3 sets and 9 

composites per set) for the F0 generation and a total of 18 composites (3 replicates per concentration 

group and 1 composite per replicate among the six concentration groups) for the F1 generation on day 0. 

On day 7, there are 3 composite weight data per replicate for 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 Lindane 

Data Files and data sheets Endpoints Comments 

Original- Biological Data.xls:

- F0 Reproduction 

- F1 Reproduction 

- F0 Summary 

- F1 Summary 

Reformatted-: 

 SMVlindanefecunity.xls: 

- F0Reproduction 

- F1Reproduction; 

 SMVlindaneSexRatio.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. 

 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- Biological Data.xls:

One sheet for each generation and each 

day 0, 7, 14 and termination 

Reformatted-SMV lindane Growth.xls 

One sheet for each generation and each 

day 0, 7, 14 and termination 

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 three sets of 

mysids, each set with 9 

composites (5 mysids per 

composite). 

 Length are 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. 

 Genders of mysids are provided 

for day 14 and termination. 

Original- Biological Data.xls:

- F0 Reproduction Survival 

- F0 Growth Survival 

- F0 Total Survival 

- F1 Reproduction Survival 

- F1 Growth Survival 

- F1 Total Survival 

Reformatted- SMV lindane 

Survival.xls:

- F0ReproductionSurvival; 

- F0GrowthSurvival 

 Survival (reproduction) (day 

7, day 14 and termination) 

 Survival (total) (day 7, day 14 

and termination) 

 Survival endpoints are adjusted 

for the non-treatment 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 lindane data set. 

Thus the total survival at day 27 
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Data Files and data sheets Endpoints Comments 

- F0TotalSurvival 

- F1ReproductionSurvival 

- F1GrowthSurvival 

- F1TotalSurvival 

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 only made available under the oneway ANOVA in STATCHARRMS. Thus for the analyses within generations it was 

elected to use the one-way ANOVA procedure. In the one-way ANOVA Model, 

STATCHARRMS pools the results across replicate tanks (and individual mysids (if the 

data were presented for each individual mysid)) within test or control groups. The oneway 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 analysis of variance, with 

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

analyses differ. The analyses within generations compare the test group results to the 

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

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

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

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

within the F1 generation, and the analyses between the F0 and F1 generations. There is 

no test for monotonicity within the two-way 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 (oneway ANOVA), and the F0 and F1 comparison between-generation analysis (two-way 
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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 however 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 lindane 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 27). This is an underestimate of 

growth mysid mortality beyond day 14. 

 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 entries for a given brood size is zero or “NA”, the 

times for the brood was also recorded as “NA” in the original data file and they are 

treated as missing in this analysis. If the brood size is missing in either brood 1 or brood 2, 

then the sum of the two broods 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 thus 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. STATCHARRMS will 

not accommodate testing for gender effects in the one-way or two-way ANOVA models. 
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3. STATISTICAL METHODS 

Statistical analysis procedures used conform to the analysis procedure 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 

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

replicate baskets. 

3.1 Comparisons of test chemical with the control 

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

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

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

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

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

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

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

3.1.1 Outlier Screens 

 For each generation, outlier screens 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 screened, 

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

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

out including the flagged values. The list of screened values is 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. 
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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 outlier test 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 the nonparametric procedures were applied. 

3.1.5 Monotonicity of response 

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

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

significant, then that was taken as evidence of monotonicity.

3.1.6 Comparisons of test chemical to controls 

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

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

monotonically increasing or monotonically decreasing with increasing test chemical concentration. 

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

departure from monotonicity, general alternatives procedures were carried out. 

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

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

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

transformed data nonparametric methods were used. The analysis procedures fall into three categories: 

 Ordered alternatives − nonparametric 

 General alternatives − parametric 

 General alternatives − nonparametric 
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Ordered alternatives – step down procedure 

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

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

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

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

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

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

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

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

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

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

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

alternative for more details. 

General alternatives – parametric procedure 

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

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

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

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

heterogeneous within-treatment variances. 

General alternatives – Nonparametric procedure 

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

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

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

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

conducted using a modified Dunn’s test. 

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 Jonckheere-
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Terpstra 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. Analysis 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, 

assuming 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 outcomes of the preliminary tests. All test procedures described in this 

report are based on the default auto option programs in STATCHARRMS. 

3.2.1 Outlier screens 

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

the two-way mixed effects 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. 

 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. 

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. 
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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. 

3.2.5 Monotonicity Test 

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

in the two way mixed effects ANOVA. 

3.2.6 F0 generation versus F1 generation 

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

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

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

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

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

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

significantly different. 

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). 
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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 endpoints), Tables 2C and 2D (growth endpoints), and Tables 3C and 3D 

(survival endpoints). Comparisons between the F0 and F1 generations are summarized in Tables 1E 

(reproduction endpoints), 2E (growth endpoints) and 3E (survival endpoints). 
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Table 1A. Summary of Reproduction Endpoints. Laboratory: SMV, Test Chemical: Lindane, Generation: F01

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 

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 10.333 5.476 8.800 14.300 15.810 21.200 5.400 9.200 1.567 

Standard 

Dev. 0.577 1.601 2.707 2.736 0.814 1.735 1.429 1.735 0.241 

N 3 21 20 20 21 20 20 20 20 

CV (%) 5.59 29.23 30.76 19.13 5.15 8.18 26.46 18.86 15.35 

32 ng/L 

(Group 2) 

Mean 9.667 6.619 10.444 17.333 16.048 20.611 4.500 8.611 2.028 

Standard 

Dev. 0.577 2.854 2.148 4.379 1.962 1.577 1.654 1.577 0.395 

N 3 21 18 18 21 18 18 18 18 

CV(%) 5.97 43.12 20.57 25.26 12.22 7.65 36.75 18.31 19.50 

80 ng/L 

(Group 3) 

Mean 9.667 6.381 8.762 15.143 15.150 20.250 5.100 8.250 1.983 

Standard 

Dev. 0.577 2.459 3.448 5.322 1.843 1.773 0.447 1.773 0.610 

N 3 21 21 21 20 20 20 20 20 

CV(%) 5.97 38.54 39.36 35.15 12.17 8.76 8.77 21.50 30.76 

200 ng/L (Group 

4) 

Mean 9.333 6.381 7.947 14.158 15.333 20.389 5.000 8.389 1.801 

Standard 

Dev. 0.577 2.037 4.403 4.167 1.155 1.243 0.000 1.243 0.495 

N 3 21 19 19 21 18 18 18 18 

CV(%) 6.19 31.92 55.40 29.43 7.53 6.10 0.00 14.82 27.48 

500 ng/L 

(Group 5) 

Mean 9.000 8.048 10.556 18.611 14.190 19.444 5.278 7.444 2.554 

Standard 

Dev. 0.000 1.658 3.240 4.031 0.928 1.042 0.958 1.042 0.664 

N 3 21 18 18 21 18 18 18 18 

CV(%) 0.00 20.60 30.69 21.66 6.54 5.36 18.16 13.99 25.98 

1250 ng/L (Group 6) 

Mean 9.667 6.667 9.000 16.125 15.238 20.929 6.154 9.000 2.003 

Standard 

Dev. 0.577 1.958 4.290 4.938 1.221 2.056 2.193 2.121 0.547 

N 3 21 16 16 21 14 13 13 13 

CV(%) 5.97 29.37 47.66 30.62 8.01 9.82 35.63 23.57 27.28 

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: Lindane, Generation: F11

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 

Control 

(Group 1) 

Mean 9.667 6.571 9.600 15.950 14.857 20.050 5.200 8.050 1.997 

Standard 

Dev. 0.577 2.039 2.927 3.818 1.195 0.826 1.152 0.826 0.520 

N 3 21 20 20 21 20 20 20 20 

CV(%) 5.97 31.03 30.49 23.94 8.04 4.12 22.15 10.26 26.05 

32 ng/L 

(Group 2) 

Mean 8.667 6.048 8.150 14.300 16.571 20.944 4.778 8.944 1.699 

Standard 

Dev. 1.155 2.655 3.631 4.041 2.271 1.474 1.215 1.474 0.484 

N 3 21 20 20 21 18 18 18 18 

CV(%) 13.32 43.90 44.56 28.26 13.70 7.04 25.44 16.48 28.45 

80 ng/L 

(Group 3) 

Mean 10.667 4.810 7.100 11.900 16.667 21.895 5.263 9.895 1.269 

Standard 

Dev. 1.155 1.940 3.177 4.051 1.461 1.629 1.147 1.629 0.416 

N 3 21 20 20 21 19 19 19 19 

CV(%) 10.83 40.33 44.75 34.04 8.76 7.44 21.79 16.47 32.76 

200 ng/L 

 (Group 4) 

Mean 9.667 3.947 5.313 9.375 16.737 21.563 4.938 9.563 1.004 

Standard 

Dev. 0.577 1.353 3.005 3.263 2.232 2.097 0.250 2.097 0.358 

N 3 19 16 16 19 16 16 16 16 

CV(%) 5.97 34.27 56.56 34.81 13.34 9.72 5.06 21.93 35.66 

500 ng/L 

(Group 5) 

Mean 10.000 4.700 10.632 15.421 15.650 21.105 5.474 9.105 1.772 

Standard 

Dev. 1.000 1.838 4.512 5.102 2.033 2.283 1.429 2.283 0.613 

N 3 20 19 19 20 19 19 19 19 

CV(%) 10.00 39.11 42.44 33.09 12.99 10.82 26.10 25.07 34.59 

1250 ng/L (Group 6) 

Mean 10.667 5.300 7.533 13.200 16.000 21.167 5.417 9.167 1.620 

Standard 

Dev. 0.577 3.080 4.779 4.648 1.453 1.801 1.443 1.801 0.583 

N 3 20 15 15 19 12 12 12 12 

CV(%) 5.41 58.11 63.44 35.21 9.08 8.51 26.65 19.64 35.97 

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

 Chemical: Lindane, Generation: F0 

Parameters 

Normality 

Test1

Homogeneity 

of Variance 

Test2

Monotonicity 

Test3

Concentration 

Trend Tests4

Overall 

Concentration 

Effect5

Pairwise 

Comparisons 

to Control6,7

Time to Mature Normal Homogeneous NonMonotonic NA No NA 

Number 

Offspring 

(brood 1)

NonNormal NA Monotonic 

Increasing trends 

for the two 

highest 

concentration 

groups 

Yes NA 

Number 

Offspring 

(brood 2)

Normal Homogeneous NonMonotonic NA No NA 

Number 

Offspring 

(two broods)

Normal Homogeneous NonMonotonic NA Yes Group 5 

Days to 

Brood 1 

NonNormal NA NonMonotonic NA Yes Groups 3, 5 

and 6 

Days to 

Brood 2 

NonNormal NA NonMonotonic NA Yes Groups 3 and 

5 

Days Between 

Broods 1 and 2 

NonNormal NA NonMonotonic NA No NA 

Total Reproductive 

Days for the First 

Two Broods 

NonNormal NA NonMonotonic NA Yes Groups 3 and 

5 

# of Offspring Per 

Female Per 

Reproduction Day 

 for the First 

Two Broods8

Normal Homogeneous Monotonic 

Increasing trends 

for the two 

highest 

concentration 

groups 

Yes NA 

% of Females 

Productively 

Active 

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

 Chemical: Lindane, 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 NonNormal NA Non-Monotonic NA No NA 

Number Offspring 

(brood 1)

NonNormal NA Non-Monotonic NA Yes Groups 4 and 5 

Number Offspring 

(brood 2) Normal Homogeneous NonMonotonic NA Yes Group 4 

Number Offspring 

(two broods) Normal Homogeneous Non-Monotonic NA Yes Groups 3 and 4 

Days to Brood 1 NonNormal NA Non-Monotonic NA Yes Groups 2, 3 and 4 

Days to Brood 2 NonNormal NA Non-Monotonic NA Yes Group 3 

Days Between Broods 1 

and 2 

NonNormal NA Non-Monotonic NA No NA 

Total Reproductive Days 

for the First Two Broods 

NonNormal NA Non-Monotonic NA Yes Group 3 

# of Offspring Per Female 

Per Reproduction Day 

 for the First 

Two Broods 

Normal Homogeneous Non-Monotonic NA Yes Groups 3 and 4 

% of Females 

Reproductively Active 

NonNormal NA NonMonotonic NA No NA 

Sex Ratio Normal Homogeneous Non-Monotonic 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. 

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

 SMV, Test Chemical: Lindane. 

Parameters 

Normality 

Test1

Homogeneity 

of Variance 

Test2

Parametric/ 

Nonparametric 

Concentration 

Main Effect 

Generation Main 

Effect 

Concentration 

by Generation 

Interaction 

Groups with 

Significant 

Difference 

between 

Generations3

Time to 

Mature Normal Homogeneous Parametric No No No NA 

Number 

Offspring 

(brood 1

Normal Homogeneous Parametric No Yes Yes Groups 4 and 

5 

Number 

Offspring 

(brood 2)

Normal Homogeneous Parametric Yes Yes No Group 4 

Number 

Offspring 

(two broods)

Normal Homogeneous Parametric Yes Yes No Groups 3, 4, 

and 5 

Days to 

Brood 1 Normal Homogeneous Parametric Yes Yes Yes Groups 3, 4, 

and 5 

Days to 

Brood 2 Normal Homogeneous Parametric No Yes No Groups 3 

and 5 

Days Between 

Broods 1 and 

2 

Normal Homogeneous Parametric Yes No No NA 

Total 

Reproductive 

Days for the 

First Two 

Broods 

Normal Homogeneous Parametric No Yes No Groups 3 and 

5 

# of Offspring 

Per Female Per 

Reproduction 

Day 

 for the First 

Two Broods 

Normal Homogeneous Parametric No Yes Yes Groups 3, 4 

and 5 

% of Females 

Productively 

Active 

NonNormal NA Nonparametric No No No NA 

Sex Ratio 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. 

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

 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) 

Length 

(mm) 

(day 7) 

Length 

(mm) 

(day 14) 

Length 

(mm) 

(Termination) 

Control 

(Group 1) 

Mean 0.035 0.297 0.642 1.098 1.843 4.458 6.303 7.421 

Standard 

Dev. 0.010 0.052 0.142 0.267 0.129 0.415 0.317 0.326 

N 27 9 32 39 135 18 32 39 

CV(%) 28.21 17.44 22.20 24.30 7.01 9.31 5.03 4.40 

32 ng/L 

(Group 2) 

Mean 0.357 0.616 1.188 4.466 6.568 7.488 

Standard 

Dev. 0.050 0.156 0.280 0.255 0.412 0.361 

N 9 38 35 17 38 35 

CV(%) 14.00 25.41 23.55 5.71 6.27 4.82 

80 ng/L 

(Group 3) 

Mean 0.281 0.675 1.069 4.670 6.636 7.370 

Standard 

Dev. 0.079 0.207 0.295 0.536 0.561 0.377 

N 9 36 48 17 36 48 

CV(%) 28.01 30.60 27.62 11.48 8.46 5.12 

200 ng/L (Group 4) 

Mean 0.316 0.891 1.047 4.791 6.815 7.368 

Standard 

Dev. 0.054 0.191 0.254 0.421 0.335 0.421 

N 9 31 41 18 31 41 

CV(%) 17.17 21.47 24.24 8.78 4.91 5.71 

500 ng/L 

(Group 5) 

Mean 0.359 0.997 1.288 4.838 6.971 7.662 

Standard 

Dev. 0.075 0.190 0.312 0.360 0.302 0.406 

N 9 31 32 18 31 32 

CV(%) 20.80 19.10 24.19 7.45 4.33 5.30 

1250 ng/L 

(Group 6) 

Mean 0.304 1.031 1.018 4.583 6.582 7.385 

Standard 

Dev. 0.058 0.254 0.322 0.229 0.501 0.451 

N 9 32 23 18 32 23 

CV(%) 19.13 24.64 31.63 5.00 7.62 6.11 

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 are 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: Lindane, 

 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 0.029 0.380 0.805 1.046 1.633 4.715 6.388 7.403 

Standard 

Dev. 0.005 0.142 0.178 0.238 0.125 0.260 0.311 0.243 

N 3 9 18 37 15 18 18 37 

CV(%) 17.56 37.49 22.11 22.76 7.66 5.52 4.87 3.28 

32 ng/L 

(Group 2) 

Mean 0.030 0.383 0.783 1.022 1.671 4.622 6.333 7.351 

Standard 

Dev. 0.011 0.149 0.174 0.191 0.064 0.433 0.171 0.298 

N 3 9 16 38 15 18 16 38 

CV(%) 35.28 38.90 22.25 18.70 3.84 9.36 2.69 4.05 

80 ng/L 

(Group 3) 

Mean 0.027 0.294 0.588 0.978 1.627 4.644 6.111 7.230 

Standard 

Dev. 0.007 0.075 0.152 0.220 0.097 0.444 0.268 0.330 

N 3 9 26 37 15 18 26 37 

CV(%) 25.70 25.44 25.85 22.54 5.95 9.56 4.39 4.57 

200 ng/L (Group 

4) 

Mean 0.033 0.307 0.622 0.986 1.705 4.544 6.058 7.124 

Standard 

Dev. 0.010 0.077 0.186 0.245 0.062 0.570 0.347 0.318 

N 3 9 24 33 15 18 24 33 

CV(%) 30.79 24.96 29.94 24.86 3.64 12.54 5.73 4.47 

500 ng/L 

(Group 5) 

Mean 0.025 0.337 0.788 1.137 1.689 4.863 6.474 7.673 

Standard 

Dev. 0.011 0.065 0.144 0.271 0.070 0.422 0.341 0.475 

N 3 9 18 30 15 18 18 30 

CV(%) 43.48 19.42 18.29 23.79 4.16 8.67 5.27 6.19 

1250 ng/L 

(Group 6) 

Mean 0.025 0.318 0.688 0.937 1.699 4.334 6.547 7.220 

Standard 

Dev. 0.009 0.035 0.171 0.246 0.094 0.400 0.543 0.522 

N 3 9 16 21 15 18 16 21 

CV(%) 38.32 11.16 24.84 26.29 5.52 9.23 8.29 7.23 

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

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

on day 14 and termination. 

3. Summaries are based on untransformed data with no screened outliers removed. 
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Table 2C. Statistical Analysis Results for Growth Endpoints. Laboratory: SMV, Test 

 Chemical: Lindane, 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)

Normal Homogeneous NonMonotonic NA No NA 

Dry 

Weight(mg) 

(day 14)

Normal Heterogeneous Monotonic 

Increasing trend 

for the three 

highest 

concentration 

groups 

Yes NA 

Dry Weight 

(mg) 

(Termination)

NonNormal NA NonMonotonic NA Yes Group 5 

Length (mm) 

(day 7)

NonNormal NA Monotonic No Yes NA 

Length (mm) 

(day 14)

NonNormal NA NonMonotonic NA Yes Groups 2, 3, 4, 

5 and 6 

Length (mm) 

(Termination) Normal Homogeneous NonMonotonic NA Yes Group 5 

1. Shapiro-Wilk test for normality. 

2. Levene Test for Homogeneity of Variances. 

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

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

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

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

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

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

conducted: 

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

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

 
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Intra-Laboratory Analysis Report for Mysid: SMV Lindane Data 

21 

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

 Chemical: Lindane, 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)

Normal Homogeneous NonMonotonic NA No NA 

Dry Weight 

(mg) 

(day 7)

Normal8

 Homogeneous NonMonotonic NA No NA 

Dry Weight 

(mg) 

(day 14)

Normal Homogeneous NonMonotonic NA Yes Groups 3 and 

4 

Dry Weight 

(mg) 

(Termination)

Normal9

 Homogeneous NonMonotonic NA No NA 

Length (mm) 

(day 0)

NonNormal NA NonMonotonic No No NA 

Length (mm) 

(day 7) Normal Heterogeneous NonMonotonic NA Yes Group 6 

Length (mm) 

(day 14) Normal Heterogeneous NonMonotonic NA Yes Groups 3 and 

4 

Length (mm) 

(Termination)

NonNormal NA NonMonotonic NA Yes Groups 4 and 

5 

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. After log-transformed, normal and homogeneous variance assumptions hold 

9. After log-transformed, normal and homogeneous variance assumptions hold. 

 
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Lindane Data 

22 

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

 Test Chemical: Lindane. 

Parameters 

Normality 

Test1

Homogeneity 

of Variance 

Test2

Parametric/ 

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 No No NA 

Dry Weight 

(mg) 

(day 14)

Normal Homogeneous Parametric Yes Yes Yes Groups 1, 2, 

4, 5 and 6 

Dry Weight 

(mg) 

(Termination)

Normal Homogeneous Parametric Yes Yes No Groups 2 and 

5 

Length (mm) 

(day 7) Normal Homogeneous Parametric No No No NA 

Length (mm) 

(day 14) Normal Homogeneous Parametric No Yes Yes Groups 3, 4 

and 5 

Length (mm) 

(Termination) Normal Homogeneous Parametric Yes 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 Lindane Data 

23 

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

 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 96.667 89.027 75.784 96.667 89.638 75.784 

Standard 

Dev. 5.774 6.208 9.413 5.774 3.745 9.413 

N 3 3 3 3 3 3 

CV(%) 5.97 6.97 12.42 5.97 4.18 12.42 

32 ng/L 

(Group 2) 

Mean 90.000 84.298 72.729 92.500 86.958 72.729 

Standard 

Dev. 5.000 10.658 9.940 0.000 4.747 9.940 

N 3 3 3 3 3 3 

CV(%) 5.56 12.64 13.67 0.00 5.46 13.67 

80 ng/L 

(Group 3) 

Mean 98.333 91.818 85.279 97.500 90.196 85.279 

Standard 

Dev. 2.887 3.149 4.829 2.500 4.493 4.829 

N 3 3 3 3 3 3 

CV(%) 2.94 3.43 5.66 2.56 4.98 5.66 

200 ng/L 

(Group 4) 

Mean 88.333 87.895 80.370 89.167 83.001 80.370 

Standard 

Dev. 2.887 3.201 8.340 5.204 8.696 8.340 

N 3 3 3 3 3 3 

CV(%) 3.27 3.64 10.38 5.84 10.48 10.38 

500 ng/L 

(Group 5) 

Mean 95.000 86.619 76.111 94.167 85.817 76.111 

Standard 

Dev. 5.000 6.659 3.469 3.819 11.398 3.469 

N 3 3 3 3 3 3 

CV(%) 5.26 7.69 4.56 4.06 13.28 4.56 

1250 ng/L 

(Group 6) 

Mean 95.000 86.475 69.697 95.000 86.608 69.697 

Standard 

Dev. 5.000 6.523 13.887 5.000 1.573 13.887 

N 3 3 3 3 3 3 

CV(%) 5.26 7.54 19.92 5.26 1.82 19.92 

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 Lindane Data 

24 

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

 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 97.778 95.956 92.308 95.833 91.667 92.308 

Standard 

Dev. 3.849 3.507 7.692 4.167 7.217 7.692 

N 3 3 3 3 3 3 

CV(%) 3.94 3.65 8.33 4.35 7.87 8.33 

32 ng/L 

(Group 2) 

Mean 93.333 88.487 77.835 91.667 87.247 77.835 

Standard 

Dev. 6.667 4.846 3.836 4.167 3.794 3.836 

N 3 3 3 3 3 3 

CV(%) 7.14 5.48 4.93 4.55 4.35 4.93 

80 ng/L 

(Group 3) 

Mean 97.778 97.917 89.744 98.611 97.222 89.744 

Standard 

Dev. 3.849 3.608 11.750 2.406 2.406 11.750 

N 3 3 3 3 3 3 

CV(%) 3.94 3.69 13.09 2.44 2.47 13.09 

200 ng/L 

(Group 4) 

Mean 100.000 100.000 86.931 98.611 97.101 86.931 

Standard 

Dev. 0.000 0.000 5.250 2.406 5.020 5.250 

N 3 3 3 3 3 3 

CV(%) 0.00 0.00 6.04 2.44 5.17 6.04 

500 ng/L 

(Group 5) 

Mean 100.000 94.118 85.769 95.833 88.889 85.769 

Standard 

Dev. 0.000 5.882 9.398 7.217 6.365 9.398 

N 3 3 3 3 3 3 

CV(%) 0.00 6.25 10.96 7.53 7.16 10.96 

1250 ng/L 

(Group 6) 

Mean 95.556 91.667 66.667 89.683 86.905 66.667 

Standard 

Dev. 3.849 3.608 10.497 5.401 1.031 10.497 

N 3 3 3 3 3 3 

CV(%) 4.03 3.94 15.75 6.02 1.19 15.75 

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 Lindane Data 

25 

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

 Chemical: Lindane, 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 NonMonotonic NA No NA 

Reproduction 

Survival Rate 

(%) 

(day 14)

Normal Homogeneous NonMonotonic NA No NA 

Reproduction 

Survival Rate 

(%) 

(Termination)

Normal Homogeneous NonMonotonic NA No NA 

Total 

Survival Rate 

(%) 

(day 7)

Normal Homogeneous NonMonotonic NA No NA 

Total 

Survival Rate 

(%) (day 14)

Normal Homogeneous NonMonotonic NA No NA 

Total 

Survival Rate 

(%) 

(Termination)

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. 
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Lindane Data 

26 

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

 Chemical: Lindane, Generation: F1 

Parameters 

Normality 

Test1

Homogeneity 

of Variance 

Test2

Monotonicity 

Test 

Concentration 

Trend Tests 

Overall 

Concentration 

Effect 

Significant 

Pairwise 

Comparisons 

to Control3

Reproduction 

Survival Rate 

(%) 

(day 7)

Normal Homogeneous NonMonotonic NA No NA 

Reproduction 

Survival Rate 

(%) 

(day 14)

Normal Homogeneous NonMonotonic NA Yes No 

Reproduction 

Survival Rate 

(%) 

(Termination)

Normal Homogeneous Monotonic 

Decreasing for 

the highest 

concentration 

group 

Yes NA 

Total 

Survival Rate 

(%) 

(day 7)

Normal Homogeneous NonMonotonic NA No NA 

Total 

Survival Rate 

(%) (day 14)

Normal Homogeneous NonMonotonic NA Yes No 

Total 

Survival Rate 

(%) 

(Termination)

Normal Homogeneous NonMonotonic NA Yes No 

1. Shapiro-Wilk test for normality. 

2. Levene Test for Homogeneity of Variances. 

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

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

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

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

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

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

conducted: 

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

within-treatment variances for normal distributed data 

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

 
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Lindane Data 

27 

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

 Chemical: Lindane. 

Parameters 

Normality 

Test1

Homogeneity 

of Variance 

Test2

Parametric/ 

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 Yes No Group 4 

Reproduction 

Survival Rate 

(%) 

(day 14)

Normal Homogeneous Parametric No Yes No Group 4 

Reproduction 

Survival Rate 

(%) 

(Termination)

Normal Homogeneous Parametric Yes Yes No Group 1 

Total 

Survival Rate 

(%) 

(day 7)

Normal Homogeneous Parametric No No No NA 

Total 

Survival Rate 

(%) (day 14)

Normal Homogeneous Parametric No Yes No Group 4 

Total 

Survival Rate 

(%) 

(Termination)

Normal Homogeneous Parametric Yes 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 Lindane Data 

28 

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

and Group For Chemical Lindane From Lab SMV. 

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

Generation and Group For Chemical Lindane From Lab SMV. 

Time To Mature (Days)

8.0

8.5

9.0

9.5

10.0

10.5

11.0

11.5

12.0

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Group

Generation F0 Generation F1

Number Offspring (brood 1)

3

4

5

6

7

8

9

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Group

Generation F0 Generation F1
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Lindane Data 

29 

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

Generation and Group For Chemical Lindane From Lab SMV. 

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

By Generation and Group For Chemical Lindane From Lab SMV. 

Number Offspring (brood 2)

4

5

6

7

8

9

10

11

12

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Group

Generation F0 Generation F1

Number Offspring (two broods)

8

9

10

11

12

13

14

15

16

17

18

19

20

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Group

Generation F0 Generation F1
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Lindane Data 

30 

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

and Group For Chemical Lindane From Lab SMV. 

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

and Group For Chemical Lindane From Lab SMV. 

Days to Brood 1

13

14

15

16

17

18

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Group

Generation F0 Generation F1

Days to Brood 2

19

20

21

22

23

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Group

Generation F0 Generation F1
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Lindane Data 

31 

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

Generation and Group For Chemical Lindane From Lab SMV. 

Figure 8. Means ± Standard Errors of Untransformed data: Total Reproductive Days for the 

First Two Broods By Generation and Group For Chemical Lindane From Lab 

SMV. 

Days Between Broods 1 and 2

4

5

6

7

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Group

Generation F0 Generation F1

Total Reproductive Days

7.0

7.5

8.0

8.5

9.0

9.5

10.0

10.5

11.0

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Group

Generation F0 Generation F1
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Lindane Data 

32 

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

per Reproductive Day for the First Two Broods By Generation and Group For 

Chemical Lindane From Lab SMV. 

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

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

# Offspring per Female per Reproductive Day 0.5

1.0

1.5

2.0

2.5

3.0

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Group

Generation F0 Generation F1

% of Females Productively Active

90

91

92

93

94

95

96

97

98

99

100

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Group

Generation F0 Generation F1
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Lindane Data 

33 

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

Group For Chemical Lindane From Lab SMV. 

Figure 12. Means ± Standard Errors of Untransformed data: Dry Weight (mg) (day 0) By 

Generation and Group For Chemical Lindane From Lab SMV. 

Sex Ratio

0.4

0.6

0.8

1.0

1.2

1.4

1.6

1.8

2.0

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Group

Generation F0 Generation F1

Dry Weight (mg)(day 0)

0.018

0.020

0.022

0.024

0.026

0.028

0.030

0.032

0.034

0.036

0.038

0.040

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Group

Generation F0 Generation F1
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Lindane Data 

34 

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

Generation and Group For Chemical Lindane From Lab SMV. 

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

Generation and Group For Chemical Lindane From Lab SMV. 

Dry Weight (mg)(day 7)

0.24

0.26

0.28

0.30

0.32

0.34

0.36

0.38

0.40

0.42

0.44

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Group

Generation F0 Generation F1

Dry Weight (mg)(day 14)

0.5

0.6

0.7

0.8

0.9

1.0

1.1

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Group

Generation F0 Generation F1
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Lindane Data 

35 

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

By Generation and Group For Chemical Lindane From Lab SMV. 

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

Generation and Group For Chemical Lindane From Lab SMV. 

Dry Weight (mg)(Termination)

0.88

0.92

0.96

1.00

1.04

1.08

1.12

1.16

1.20

1.24

1.28

1.32

1.36

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Group

Generation F0 Generation F1

Length (mm)(day 0)

1.60

1.62

1.64

1.66

1.68

1.70

1.72

1.74

1.76

1.78

1.80

1.82

1.84

1.86

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Group

Generation F0 Generation F1
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Lindane Data 

36 

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

Generation and Group For Chemical Lindane From Lab SMV. 

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

Generation and Group For Chemical Lindane From Lab SMV. 

Length (mm)(day 7)

4.2

4.3

4.4

4.5

4.6

4.7

4.8

4.9

5.0

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Group

Generation F0 Generation F1

Length (mm)(day 14)

5.9

6.0

6.1

6.2

6.3

6.4

6.5

6.6

6.7

6.8

6.9

7.0

7.1

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Group

Generation F0 Generation F1
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Lindane Data 

37 

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

Generation and Group For Chemical Lindane From Lab SMV. 

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

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

Length (mm)(Termination)

7.0

7.1

7.2

7.3

7.4

7.5

7.6

7.7

7.8

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Group

Generation F0 Generation F1

Reproduction Survival Rate (%)(day 7)

86

88

90

92

94

96

98

100

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Group

Generation F0 Generation F1
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Lindane Data 

38 

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

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

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

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

Reproduction Survival Rate (%)(day 14)

70

75

80

85

90

95

100

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Group

Generation F0 Generation F1

Reproduction Survival Rate (%)(Termination)

60

65

70

75

80

85

90

95

100

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Group

Generation F0 Generation F1
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Lindane Data 

39 

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

By Generation and Group For Chemical Lindane From Lab SMV. 

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

By Generation and Group For Chemical Lindane From Lab SMV. 

Total Survival Rate (%)(day 7)

86

88

90

92

94

96

98

100

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Group

Generation F0 Generation F1

Total Survival Rate (%)(day 14)

76

80

84

88

92

96

100

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Group

Generation F0 Generation F1
DRAFT 

Intra-Laboratory Analysis Report for Mysid: SMV Lindane Data 

40 

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

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

Total Survival Rate (%)(Termination)

68

72

76

80

84

88

92

96

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Control 32

ng/L

80

ng/L

200

ng/L

500

ng/L

1250

ng/L

Group

Generation F0 Generation F1