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Intra-Laboratory Analysis Report for Mysid: UNT 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 UNT 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 .......................................................................................................... 8 

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

3.1.1 Outlier screens ..................................................................................................... 8 

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

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

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

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

3.1.6 Comparisons of test chemical to controls .......................................................... 10 

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

3.2.1 Outlier screens ................................................................................................... 12 

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

3.2.3 Transformation of data ...................................................................................... 13 

3.2.4 Heterogeneous variances ................................................................................... 13 

3.2.5 Monotonicity Test .............................................................................................. 13 

3.2.6 F0 generation versus F1 generation ................................................................... 13 

4. Results ........................................................................................................................... 13 

Tables

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

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

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

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

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

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

Chemical: Lindane, Generation: F0 .............................................................................................. 17 
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Table 1D. Statistical Analysis Results for Reproduction Endpoints. Laboratory: UNT, Test 

Chemical: Lindane, Generation: F1 .............................................................................................. 18 

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

UNT, Test Chemical: Lindane. .................................................................................................... 19 

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

Generation: F01,2,3,4,5 ...................................................................................................................... 20 

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

Generation: F11,2,3,4 ........................................................................................................................ 21 

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

Chemical: Lindane, Generation: F0 .............................................................................................. 22 

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

Chemical: Lindane, Generation: F1 .............................................................................................. 23 

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

Test Chemical: Lindane. ................................................................................................................ 24 

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

Generation: F01,2,3 .......................................................................................................................... 25 

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

Generation: F11,2,3 .......................................................................................................................... 26 

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

Chemical: Lindane, Generation: F0 .............................................................................................. 27 

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

Chemical: Lindane, Generation: F1 .............................................................................................. 28 

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

Test Chemical: Lindane. ................................................................................................................ 29 

Figures

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

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

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

Lindane From Lab UNT. ...................................................................................... 30 

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

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

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

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

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

From Lab UNT. .................................................................................................... 32 
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Figure 6. Days to Brood 2 By Generation and Group For Chemical Lindane 

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

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

Lindane From Lab UNT. ...................................................................................... 33 

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

Chemical Lindane From Lab UNT. ...................................................................... 33 

Figure 9. Sex Ratio By Generation and Group For Chemical Lindane From Lab UNT. ..... 34 

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

Lindane From Lab UNT. ...................................................................................... 34 

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

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

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

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

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

Lindane From Lab UNT. ...................................................................................... 37 

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

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

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

From Lab UNT. .................................................................................................... 38 

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

From Lab UNT. .................................................................................................... 38 

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

Lindane From Lab UNT. ...................................................................................... 40 

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

Chemical Lindane From Lab UNT. ...................................................................... 40 

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

Chemical Lindane From Lab UNT. ...................................................................... 41 

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

Group For Chemical Lindane From Lab UNT. .................................................... 41 

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

Lindane From Lab UNT. ...................................................................................... 43 

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

Lindane From Lab UNT. ...................................................................................... 43 

 
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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 UNT 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 29 (F0) and Day 30 

(F1) for UNT), mysids from the reproduction baskets are also measured for weights and lengths. 
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Intra-laboratory statistical analysis of the UNT 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)1

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

and 14 (total survival, per tank) 

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

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

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

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

due to the absence of appropriate data. 

 

1

 The reproduction termination survival rates were based only on the14 adult reproduction mysids in the seven brood 

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

accounted for in the survival rates. 
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(iii) Growth (F0, F1) (8)2, 3 

a. Length at day 0 (per mysid) 

b. Length at day 7 (per mysid) 

c. Length at day 14 (per mysid) 

d. Length at termination. (per mysid) 

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

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

g. Weight at day 14 (per mysid) 

h. Weight at termination (per mysid). 

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

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

concentration as their parents’ treatment groups. 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 (5 mysids per composite, 9 composites per 

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

are 3 composite (2 mysids per composite) weight data 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. 

 

 

2

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

3

 Weight and length results were reported pooled across replicate tanks within control or test concentration groups. 

Tank identities were not provided in the datasheets. 
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Table 0. Data information for UNT Lindane 

Data Files and data sheets Endpoints Comments 

Original - Lindane UNT Excel 

Spreadsheet.xlsx:

− Time to First Brood Release 

for F0 and F1 generations 

− Time to Second Brood 

Release for F0 and F1 

generations 

− Interbrood Duration 

− Time to Maturation 

− Reproduction Fo 

− Reproduction F1 

− Sex Ratio Fo 

− Sex Ratio F1 

Reformatted – 

 UNTlindanefecunity.xlsx: 

- One datasheet per endpoint 

per generation 

 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 

 Sex ratio 

The following two variables are 

not provided: 

 Offspring per female per 

reproductive day for the first 

two broods 

 Total reproductively active 

days for the first two broods 

 If females died before the first or 

second brood, the 0 brood sizes were 

recorded in the original data files. Per 

EPA suggestion, such data are changed 

as: zero if female is still living, and 

missing for a brood (1st or 2nd brood) if 

female died without releasing the 

brood. The total offspring for the first 2 

broods are therefore recalculated and 

displayed in the reformatted data. % of 

female reproductively active, and sex 

ratios are reported per tank. The other 

reproductive endpoints are reported per 

mating pair. 

 Replicate IDs are given (labeled as 1, 2, 

3). 

 Sex ratios were defined as % of females 

over all animals in the original data file. 

They were redefined as 

#males/#females in the reformatted 

data. 

Original- Lindane UNT Excel 

Spreadsheet.xlsx:

− Fo,F1 Lengths Day 0 

− Fo Lengths Day 7 

− Fo Lengths Day 14 

− Fo Lengths Termination 

− F1 Lengths Day 7 

− F1 Lengths Day 14 

− F1 Lengths Termination 

− Fo,F1 Weight Day 0 

− Fo Weight Day 7 

− Fo Weight Day 14 

− Fo Weight Termination 

− F1 Weight Day 7 

− F1 Weight Day 14 

− F1 Weight Termination 

Reformatted-UNT lindane 

Growth.xls 

One sheet for each generation 

and each of days 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. 

 Lengths are reported per mysid. 

 Weights are reported by mysid 

composites per replicate tank. For F0, 

day 0 9 composites (5 mysids per 

composite). For F1, day 0 nine 

composites per concentration group (, 5 

mysids per composite).For day 7 three 

composites per replicate and two 

mysids per composite), and per mysid 

for day 14 and for termination. 

 Lengths are measured per mysid at all 

times, from day 0 to termination. 

 Replicate (basket) ID’s are not 

available, thus we are not able to 

include a random replicate effect into 

the two-way ANOVA model (when 

comparing the two generations in 

STATCHARRMS). 

 No gender information is provided for 

day 14 and termination. We do not 

analyze the data by gender. 

Original- Survival and 

Reproduction Lindane.xlsx 

- F0 Survival and Repro. 

- F1 Survival and Repro. 

 Survival (reproduction) (day 

7, day 14 and termination) 

 Survival (total) (day 7, day 

 Reformatted survival data were hand 

entered using the information from the 

original files. The original files have 

survival data from day 0 to day 14. 
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Data Files and data sheets Endpoints Comments 

Reformatted- UNT Lindane 

Survival

- F0Survival 

- F1Survival 

14) 

 Survival (total) at 

termination is not available4

. 

Thus the survival data can be 

determined using the information on 

days 7 and 14. 

 The initial numbers of mysids are 20 

for F0 and 15 for F1 per tank. Thus 

there should never be more than 20 

mysids in a tank. Sometimes there are 

more than 20 mysids reported alive on 

day 7, we treated them as 20.

 Sometimes more mysids survived at a 

later date than at a previous date. We 

will determine the number based on the 

information. For example, on day 7, the 

number of survivals was recorded as 19 

but there were 20 survivals on day 14. 

We will assume survival number on day 

7 as 20 instead of 19.

 There are no survival records for 

mysids, that were not assigned to the 

brood chambers after day 14. The 

survival for termination is determined 

using the animals from the brood 

chambers only. This is reproduction 

survival. 

 If there is a male death, which is 

assumed if number of animals goes 

from 2 to 1 and then back to 2, one addin animal should be included in the 

survival pool. If there is a female death, 

which is assumed if number of animals 

goes from 2 to 1 and then stops (study 

for the brood chamber was terminated), 

one dead and one live are counted. 

 If the data for the two animals in a 

breeding pairs ended at the same time 

before the terminal day, they are 

assumed to be dead 

 Survival endpoints are reported per 

replicate tank. 

2.3 Analysis Considerations 

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

 STATCHARRMS uses totally independent statistical procedures for analyses within the 

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

 

4

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

numerator values are inconsistent with survival information in the Survival and Reproduction spreadsheet. The 

denominator values are not adjusted for non-treatment related mortality, in particular for the growth mysids that are 

removed at day 7 and day 14 for determination of lengths and weights. 
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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 procedure, 

STATCHARRMS pools the results across test or control group. The one-way ANOVA 

(parametric or nonparametric forms) was applied to the one generation data and there are 

no random replicate effects. For the comparison between the F0 and F1 generations 

STATCHARRMS utilizes a two-way mixed effects analysis of variance, with replicate 

tanks within treatment groups treated as random effects. For some endpoints, if the 

replicate information is not available the random effect was not included in the model. 

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

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

results between generations, either pooled across groups or separately within test or 

control groups. 

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

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

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

no test for monotonicity within the two-way AVOVA for comparisons between 

generations. The results of the preliminary tests may differ among the F0 analysis, the F1 

analysis, and the analysis for comparison between F0 and F1. Thus the approaches for 

the principal tests used are independently determined among these analyses. For example, 

for a particular endpoint some of 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 analyses (two-way 

ANOVA). The statistical potential outliers are screened values based on the statistical 

model applied, and they may not be in fact biologically important. The potential outliers 

from both one-way and two-way ANOVA’s should be jointly considered. EPA will 

determine whether the flagged values should be included in the subsequent analyses or 

should be treated as outliers. 

 The statistical analysis plan 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 
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analyses. The screened values however were enumerated in the STATCHARRMS 

outputs and will be sent to EPA for review. 

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

assigned to or later added into the brood chambers. This accounts for lethality only 

within the seven breeding pairs from day 14 to termination day. It does not account for 

lethality among the reproductive mysid prior to being paired at day 14. 

 Total survival at termination is not determined. The F0 survival and F1 survival 

worksheets in the Lindane UNT Excel spreadsheet are not used in the analysis because 

they are inconsistent with lethality shown in the Survival and Reproduction Lindane 

spreadsheet. The number of dead mysids per tank is not consistent with the number of 

mortalities shown in the Survival and Reproduction Lindane spreadsheet, which is taken 

to be authoritative with respect to survival. The denominators are about 40 in the F0 

survival worksheets and about 30 for the F1 survival worksheets. These values do not 

adjust for the numbers of growth or reproductive mysids that were removed from the test 

for reasons that were not test compound related (e.g. growth mysids removed on day 7 or 

day 14 for weight and length determinations or breeding pair males that were removed 

from test because the breeding pair was shut down after the female died.) Survival and 

reproduction spreadsheet survival values are based only on reproduction mysids. 

 

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

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

EPA has suggested that if the female is living the 0 brood sizes should be counted as 0 

and included in the brood size averages. If the female died before the recorded zero size 

is generated, the brood size should be counted as “NA” (or recorded as a missing). 

Battelle has changed the data entries in the columns for brood size 1 and brood size 2 in 

the Excel spreadsheet according to this suggestion. In the reformatted data, the changes 

are also made to the affected endpoints: total number of offspring in the first two broods, 

and the number of offspring per female per reproductive day during the two broods. If the 

data entries for a given brood size is 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 This applies to endpoints: number 

of offspring in brood 1, number of offspring in brood 2, number of offspring in broods 1 

and 2. 
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 For the UNT Lindane weight and length growth data the way that the data are reported 

limits the analysis procedures that can be used. The lengths and weights are reported by 

individual mysids or by mysid composite, however replicate tank information is not 

included. Thus the test group response averages are pooled across replicate tanks within 

test or control groups and effects of random variation due to replicate tank cannot be 

separated from the residual variation in the two-way analysis of variance model. 

3. STATISTICAL METHODS 

Statistical analysis procedures used conform to the statistical analysis procedures 

specified in the STATCHARRMS package. Statistical analyses include two components: 

comparisons of test chemical with the control within each generation and comparison of the F0 

and F1 generations. The statistical analysis was carried out in a univariate fashion independently 

for each endpoint. 

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

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

then the analysis 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 are to be 

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

screened, are in fact biologically unimportant. EPA will determine whether the flagged values 

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

3.1.5 Monotonicity of response 

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

from non-normal or heterogeneous data). If the linear contrast was significant but the quadratic 

contrasts was not significant, then that was taken as evidence of monotonicity.
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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 alternative for more details. 
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General alternatives – parametric procedure 

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

was not rejected, parametric general alternative procedures were used. If the overall 

concentration effect with the ANOVA was significant, comparisons of test concentrations to the 

control were conducted using Dunnett’s test with homogeneous within-treatment variances 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 non-parametric procedures 

are found appropriate. (2) There are “massive ties” in the responses, i.e., 25% or more of the 

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

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

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

3.2 Comparisons of the F0 and F1 generations 

 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 
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between the two generations. The residual variance was treated as homogeneous between 

generations if Levene’s test accepts a homogeneous variances. Otherwise a heterogeneous 

residual variance is used. For the length and weight responses replicate tank information was not 

provided in the dataset. Therefore variation due to replicate tank could not be separated from 

residual variation within tanks. Thus repeated measures analyses could not be carried out. 

 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 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 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 and will be sent to EPA for 

review. 

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 (for the growth measurements, 

random effects was not assumed due to the lack of replicate information). 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), 

where the residuals were from the model fit described in the normality test (Section 3.2.2). 

Outliers were included in the test. 

3.2.5 Monotonicity Test 

 A test for monotonicity was not carried out as ordered alternatives procedures are 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, Ftests 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 
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 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, Tables 2C and 2D, and Tables 3C and 3D. Comparisons 

between the F0 and F1 generations are summarized in Tables 1E, 2E and 3E. 
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Table 1A. Summary of Reproduction Endpoints. Laboratory: UNT, Test Chemical: 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 

% of Females 

Productively 

Active 

Sex Ratio 

(males/females) 

Control 

(Group 1) 

Mean 13.235 5.941 6.308 12.077 19.294 23.800 4.900 80.952 1.126 

Standard 

Dev. 

2.412 2.727 4.423 5.346 2.392 2.251 0.316 8.248 0.311 

N 17 17 13 13 17 10 10 3 3 

CV (%) 18.22 45.89 70.12 44.26 12.4 9.46 6.45 10.19 27.64 

0.032 ppb 

(Group 2) 

Mean 12.444 6.550 8.421 14.947 18.389 24.643 6.067 90.476 1.773 

Standard 

Dev. 

3.854 2.564 5.092 6.794 3.883 2.307 1.870 16.496 0.253 

N 18 20 19 19 18 14 15 3 3 

CV(%) 30.97 39.15 60.46 45.46 21.11 9.36 30.82 18.23 14.28 

0.08 ppb 

(Group 3) 

Mean 12.167 5.450 8.684 14.263 18.167 23.750 5.824 90.476 1.706 

Standard 

Dev. 

1.295 2.585 4.435 5.108 1.295 1.483 1.879 16.496 0.345 

N 18 20 19 19 18 16 17 3 3 

CV(%) 10.64 47.43 51.07 35.81 7.13 6.25 32.26 18.23 20.21 

0.2 ppb 

(Group 4) 

Mean 12.333 5.222 8.588 13.765 18.389 23.000 5.125 95.238 1.956 

Standard 

Dev. 

1.782 1.957 3.124 2.796 1.819 0.679 1.088 8.248 0.758 

N 18 18 17 17 18 14 16 3 3 

CV(%) 14.45 37.48 36.37 20.31 9.89 2.95 21.23 8.66 38.73 

0.5 ppb 

(Group 5) 

Mean 11.591 6.000 6.667 12.467 17.591 23.400 5.667 100.000 1.568 

Standard 

Dev. 

1.790 2.121 3.958 4.658 1.790 1.075 1.723 0.000 1.154 

N 22 21 15 15 22 10 12 3 3 

CV(%) 15.45 35.36 59.37 37.36 10.18 4.59 30.41 0 73.59 

1.25 ppb 

(Group 6) 

Mean 10.105 8.100 7.286 16.286 16.421 19.800 5.667 95.238 1.228 

Standard 

Dev. 

1.197 2.382 5.648 7.868 1.539 1.483 4.179 8.248 0.842 

N 19 20 7 7 19 5 6 3 3 

CV(%) 11.85 29.41 77.53 48.31 9.37 7.49 73.75 8.66 68.58 

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: UNT, 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 

% of Fe

Produc

Acti

Control 

(Group 1) 

Mean 11.357 4.867 8.143 13.286 17.214 22.818 5.545 83.3

Standard Dev. 1.447 3.204 4.435 6.592 1.477 2.040 1.214 28.8

N 14 15 14 14 14 11 11 3 

CV(%) 12.74 65.84 54.47 49.61 8.58 8.94 21.88 34.6

0.032 ppb 

(Group 2) 

Mean 10.000 5.824 7.667 13.467 16.000 21.500 5.286 100.0

Standard Dev. 1.138 1.776 3.266 3.642 1.138 1.092 1.069 0.00

N 18 17 15 15 18 14 14 3 

CV(%) 11.38 30.5 42.6 27.05 7.11 5.08 20.23 0 

0.08 ppb 

(Group 3) 

Mean 11.429 3.867 7.000 11.167 17.500 22.100 4.900 93.3

Standard Dev. 1.453 1.407 3.438 3.407 1.401 0.738 0.316 11.5

N 14 15 12 12 14 10 10 3 

CV(%) 12.71 36.4 49.11 30.51 8 3.34 6.45 12.3

0.2 ppb 

(Group 4) 

Mean 11.350 4.800 7.842 12.474 17.350 23.000 5.611 90.4

Standard Dev. 1.137 1.399 4.311 4.563 1.137 1.749 1.650 8.24

N 20 20 19 19 20 18 18 3 

CV(%) 10.02 29.15 54.97 36.58 6.55 7.6 29.4 9.1

0.5 ppb 

(Group 5) 

Mean 10.882 5.294 7.917 12.833 16.882 21.125 4.700 89.6

Standard Dev. 1.965 3.312 4.010 5.024 1.965 0.835 0.675 9.01

N 17 17 12 12 17 8 10 3 

CV(%) 18.05 62.56 50.66 39.15 11.64 3.95 14.36 10.0

1.25 ppb 

(Group 6) 

Mean 10.000 8.375 7.667 17.000 16.091 20.333 5.333 83.3

Standard Dev. 1.673 2.825 4.041 4.359 1.700 1.155 0.577 28.8

N 11 8 3 3 11 3 3 3 

CV(%) 16.73 33.73 52.71 25.64 10.57 5.68 10.83 34.6

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

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

Chemical: Lindane, Generation: F0 

Parameters 

Normality 

Test1

Homogeneous 

of Variance 

Test2

Monotonicity 

Test3

Concentration 

Trend Tests4

Overall 

Concentration 

Effect5

Pairwise 

Comparisons 

to Control6,7

Time to Mature Non-Normal NA Monotonic 

Decreasing trend for 

the two highest 

concentration 

groups 

Yes NA 

Number 

Offspring 

(Brood 1)

Normal Homogeneous NonMonotonic NA Yes Group 6 

Number 

Offspring 

(Brood 2)

Non-Normal NA NonMonotonic NA No No 

Number 

Offspring 

(Two Broods)

Non-Normal NA NonMonotonic NA No NA 

Days to 

Brood 1 Non-Normal NA Monotonic 

Decreasing trend for 

the two highest 

concentration 

groups 

Yes NA 

Days to 

Brood 2 Normal8

 Heterogeneous NonMonotonic NA Yes Group 6 

Days Between Broods 

1 and 2 Non-Normal NA NonMonotonic NA No 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. LeveneTest for Homogeneity of Variances. 

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

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

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

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

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

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

conducted: 

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

within-treatment variances for normal distributed data 

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

8. Normal when Log-transformed. 
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Table 1D. Statistical Analysis Results for Reproduction Endpoints. Laboratory: UNT, Test 

Chemical: Lindane, Generation: F1 

Parameters 

Normality 

Test1

Homogeneous 

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 Yes No 

Number Offspring 

(Brood 1) Normal Heterogeneous Non-Monotonic NA Yes No 

Number Offspring 

(Brood 2)

NonNormal NA Non-Monotonic NA No NA 

Number Offspring 

(Two Broods) Normal Homogeneous Non-Monotonic NA No NA 

Days to Brood 1 NonNormal NA Non-Monotonic NA Yes No 

Days to Brood 2 NonNormal NA Monotonic NA Yes NA 

Days Between Broods 1 

and 2 

NonNormal NA Non-Monotonic NA No NA 

% 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. 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: 

UNT, Test Chemical: Lindane. 

Parameters 

Normality 

Test1

Homogeneous 

of Variance 

Test2

Parameteric/ 

Nonparametric 

Concentration 

Main Effect 

Generation Main 

Effect 

Concentration 

by Generation 

Interaction 

Groups with 

Significant 

Difference 

between 

Generations3

Time to 

Mature Normal Homogeneous Parametric Yes Yes No Group 2 

Number 

Offspring 

(Brood 1

Normal Homogeneous Parametric Yes No No NA 

Number 

Offspring 

(Brood 2)

Normal Homogeneous Parametric No No No NA 

Number 

Offspring 

(Two Broods)

Normal Homogeneous Parametric No No No NA 

Days to 

Brood 1 Normal Homogeneous Parametric No Yes No Group 2 

Days to 

Brood 2 Normal4 Homogeneous Parametric Yes Yes Yes Groups 2 and 

5 

Days Between 

Broods 1 and 

2 

Normal Homogeneous Parametric No No No NA 

% of Females 

Productively 

Active 

NonNormal NA Nonparametric No No No NA 

Sex Ratio Normal Homogeneous Parametric No Yes No No 

1. Shapiro-Wilk test for normality. 

2. Levene test for homogeneous of variances. 

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

4. Normal when log-transformed. 

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

Generation: F01,2,3,4,5

Concentration Statistics 

Dry 

Weight 

(mg) 

(day 0) 

Dry 

Weight 

(mg) 

(day 7) 

Dry Weight 

(mg) 

(day 14) 

Dry Weight 

(mg) 

(Termination) 

Length 

(mm) 

(day 0) 

Length 

(mm) 

(day 7) 

Length 

(mm) 

(day 14) 

Length 

(mm) 

(Termination) 

Control 

(Group 1) 

Mean 0.045 0.119 0.573 0.905 1.638 3.144 4.680 6.311 

Standard Dev. 0.009 0.046 0.145 0.149 0.176 0.415 0.444 0.172 

N 9 9 35 36 45 18 35 36 

CV(%) 19.61 38.1 25.21 16.43 10.76 13.19 9.48 2.73 

0.032 ppb 

(Group 2) 

Mean 0.108 0.472 1.014 3.200 4.710 6.344 

Standard Dev. 0.053 0.128 0.223 0.329 0.483 0.268 

N 9 29 42 18 29 43 

CV(%) 49.38 27.09 22.04 10.28 10.25 4.23 

0.08 ppb 

(Group 3) 

Mean 0.094 0.485 1.057 3.272 4.905 6.405 

Standard Dev. 0.036 0.166 0.218 0.304 0.589 0.282 

N 9 36 41 18 37 41 

CV(%) 38.58 34.14 20.61 9.31 12.01 4.4 

0.2 ppb 

(Group 4) 

Mean 0.072 0.481 1.048 3.383 5.211 6.239 

Standard Dev. 0.026 0.138 0.255 0.287 0.634 0.376 

N 9 27 41 18 27 41 

CV(%) 36.42 28.75 24.37 8.5 12.16 6.03 

0.5 ppb 

(Group 5) 

Mean 0.149 0.563 0.961 3.744 5.195 6.313 

Standard Dev. 0.051 0.161 0.278 0.357 0.579 0.313 

N 9 38 25 18 38 24 

CV(%) 34.57 28.52 28.92 9.53 11.15 4.95 

1.25 ppb 

(Group 6) 

Mean 0.222 0.595 1.027 4.039 5.294 6.455 

Standard Dev. 0.084 0.178 0.251 0.595 0.436 0.284 

N 9 35 11 18 35 11 

CV(%) 38.02 29.88 24.46 14.74 8.23 4.4 

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. 

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

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

Generation: F11,2,3,4

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 0.022 0.247 0.719 1.030 1.482 3.956 5.641 6.273 

Standard 

Dev. 0.003 0.041 0.125 0.245 0.101 0.340 0.197 0.303 

N 9 9 22 27 45 18 22 26 

CV(%) 12.94 16.56 17.42 23.78 6.79 8.59 3.49 4.83 

0.032 ppb 

(Group 2) 

Mean 0.023 0.234 0.716 0.972 1.482 3.772 5.574 6.110 

Standard 

Dev. 0.006 0.030 0.127 0.231 0.115 0.232 0.300 0.248 

N 9 9 23 39 45 18 23 39 

CV(%) 24.96 12.77 17.79 23.78 7.78 6.15 5.39 4.06 

0.08 ppb 

(Group 3) 

Mean 0.023 0.265 0.633 1.031 1.469 4.074 5.347 6.092 

Standard 

Dev. 0.007 0.071 0.101 0.200 0.114 0.376 0.257 0.277 

N 9 9 19 27 45 17 19 26 

CV(%) 29.6 26.87 16.04 19.39 7.79 9.23 4.8 4.55 

0.2 ppb 

(Group 4) 

Mean 0.017 0.237 0.705 0.995 1.427 4.100 5.463 6.128 

Standard 

Dev. 0.005 0.040 0.104 0.202 0.116 0.428 0.279 0.266 

N 9 9 19 29 45 18 19 29 

CV(%) 31.6 16.93 14.82 20.29 8.1 10.45 5.11 4.34 

0.5 ppb 

(Group 5) 

Mean 0.023 0.256 0.672 0.993 1.467 4.194 5.585 6.352 

Standard 

Dev. 0.007 0.038 0.089 0.192 0.138 0.335 0.208 0.333 

N 9 9 13 21 45 18 13 21 

CV(%) 29.99 14.7 13.31 19.29 9.42 7.99 3.72 5.24 

1.25 ppb 

(Group 6) 

Mean 0.022 0.291 0.618 1.007 1.462 4.150 5.525 6.711 

Standard 

Dev. 0.006 0.134 0.133 0.131 0.185 0.338 0.238 0.203 

N 9 9 8 9 45 18 8 9 

CV(%) 25.75 46.16 21.5 13.02 12.65 8.15 4.3 3.02 

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. 

4. Replicate tank within group information is not provided for the growth endpoints. 
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Table 2C. Statistical Analysis Results for Reproduction Endpoints. Laboratory: UNT, Test 

Chemical: Lindane, Generation: F0 

Parameters 

Normality 

Test1

Homogeneous of 

Variance Test2

Monotonicity 

Test3

Concentration 

Trend Tests4

Overall 

Concentration 

Effect5

Pairwise 

Comparisons to 

Control6,7

Dry Weight(mg) 

 (day 7) Normal8 Homogeneous Non-Monotonic NA Yes Group 6 

Dry Weight(mg) 

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

Dry Weight (mg) 

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

Length (mm) 

(day 7) Non-Normal NA Non-Monotonic NA Yes Groups 5 and 6 

Length (mm) 

(day 14) Non-Normal NA Monotonic 

Increasing trends for 

the four highest 

groups 

Yes NA 

Length (mm) 

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

1. Shapiro-Wilk test for normality. 

2. LeveneTest for Homogeneity of Variances. 

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

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

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

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

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

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

conducted: 

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

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

8. Normal when square root-transformed. 

 
DRAFT 

Summary Tables and Figures for Mysid: UNT Lindane Data

23 

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

Chemical: Lindane, Generation: F1 

Parameters 

Normality 

Test1

Homogeneous of 

Variance Test2

Monotonicity 

Test3

Concentration 

Trend Tests4

Overall 

Concentration 

Effect5

Pairwise 

Comparisons to 

Control6,7

Dry Weight (mg) 

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

Dry Weight (mg) 

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

Dry Weight (mg) 

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

Dry Weight (mg) 

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

Length (mm) 

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

Length (mm) 

(day 7) Normal Homogeneous Monotonic 

Increasing 

concentration trends 

for the two highest 

concentration 

groups. 

Yes NA 

Length (mm) 

(day 14) Normal Homogeneous Non-Monotonic NA Yes Group 3 

Length (mm) 

(Termination) Normal Homogeneous Non-Monotonic NA Yes Group 6 

1. Shapiro-Wilk test for normality. 

2. Levene Test for homogeneous of variances. 

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

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

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

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

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

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

conducted: 

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

within-treatment variances for normal distributed data 

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

 
DRAFT 

Summary Tables and Figures for Mysid: UNT Lindane Data

24 

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

Test Chemical: Lindane. 

Parameters 

Normality 

Test1

Homogeneous 

of Variance 

Test2

Parameteric/ 

Nonparametric 

Concentration 

Main Effect 

Generation 

Main Effect 

Concentration 

by Generation 

Interaction 

Groups with 

Significant 

Difference 

between 

Generations3

Dry Weight(mg) 

(day 7)

NonNormal NA Nonparametric Yes No Yes Group 6 

Dry Weight 

(mg) 

(day 14)

Normal Homogeneous Parametric Yes Yes Yes Groups 1, 2, 

3, 4 and 5 

Dry Weight 

(mg) 

(Termination)

NonNormal NA Nonparametric No No No NA 

Length (mm) 

(day 7) Normal Homogeneous Parametric Yes Yes Yes Groups 1, 2, 

3, 4 and 5 

Length (mm) 

(day 14)

NonNormal NA Nonparametric Yes No Yes Groups 1, 2 

and 4 

Length (mm) 

(Termination)

NonNormal NA Nonparametric Yes No Yes Groups 3 and 

6 

1. Shapiro-Wilk test for normality. 

2. Levene test for homogeneous of variances. 

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

 
DRAFT 

Summary Tables and Figures for Mysid: UNT Lindane Data

25 

Table 3A. Summary of Survival Endpoints. Laboratory: UNT, 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) 

Control 

(Group 1) 

Mean 96.667 78.333 77.302 96.667 80.392 

Standard 

Dev. 2.887 2.887 10.060 3.819 11.135 

N 3 3 3 3 3 

CV(%) 2.99 3.69 13.01 3.95 13.85 

0.032 ppb 

(Group 2) 

Mean 98.333 86.667 83.056 96.667 79.412 

Standard 

Dev. 2.887 2.887 6.989 3.819 10.605 

N 3 3 3 3 3 

CV(%) 2.94 3.33 8.41 3.95 13.35 

0.08 ppb 

(Group 3) 

Mean 98.333 88.333 71.528 99.167 90.196 

Standard 

Dev. 2.887 7.638 21.700 1.443 6.123 

N 3 3 3 3 3 

CV(%) 2.94 8.65 30.34 1.46 6.79 

0.2 ppb 

(Group 4) 

Mean 100.000 95.000 67.703 99.167 86.275 

Standard 

Dev. 0.000 8.660 10.994 1.443 7.402 

N 3 3 3 3 3 

CV(%) 0 9.12 16.24 1.46 8.58 

0.5 ppb 

(Group 5) 

Mean 95.000 80.000 64.128 97.500 86.275 

Standard 

Dev. 0.000 8.660 7.606 0.000 6.123 

N 3 3 3 3 3 

CV(%) 0 10.83 11.86 0 7.1 

1.25 ppb 

(Group 6) 

Mean 100.000 80.000 46.296 98.333 84.314 

Standard 

Dev. 0.000 18.028 9.601 1.443 8.985 

N 3 3 3 3 3 

CV(%) 0 22.53 20.74 1.47 10.66 

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

2. Survival rates were measured per tank. 

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

 
DRAFT 

Summary Tables and Figures for Mysid: UNT Lindane Data

26 

Table 3B. Summary of Survival Endpoints. Laboratory: UNT, 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) 

Control 

(Group 1) 

Mean 97.778 84.444 66.667 98.889 86.111 

Standard 

Dev. 3.849 13.878 14.434 1.925 6.365 

N 3 3 3 3 3 

CV(%) 3.94 16.43 21.65 1.95 7.39 

0.032 ppb 

(Group 2) 

Mean 100.000 95.556 82.165 98.889 91.667 

Standard 

Dev. 0.000 3.849 16.109 1.925 4.167 

N 3 3 3 3 3 

CV(%) 0 4.03 19.61 1.95 4.55 

0.08 ppb 

(Group 3) 

Mean 93.333 88.889 75.354 85.556 75.611 

Standard 

Dev. 11.547 7.698 7.044 17.105 18.097 

N 3 3 3 3 3 

CV(%) 12.37 8.66 9.35 19.99 23.93 

0.2 ppb 

(Group 4) 

Mean 100.000 93.333 70.476 100.000 93.056 

Standard 

Dev. 0.000 11.547 10.034 0.000 8.674 

N 3 3 3 3 3 

CV(%) 0 12.37 14.24 0 9.32 

0.5 ppb 

(Group 5) 

Mean 86.667 82.222 64.545 82.222 70.833 

Standard 

Dev. 11.547 10.184 18.350 3.849 11.024 

N 3 3 3 3 3 

CV(%) 13.32 12.39 28.43 4.68 15.56 

1.25 ppb 

(Group 6) 

Mean 68.889 46.667 58.333 67.778 44.444 

Standard 

Dev. 13.878 24.037 14.434 8.389 15.775 

N 3 3 3 3 3 

CV(%) 20.15 51.51 24.74 12.38 35.49 

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

2. Survival rates were measured per tank. 

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

Summary Tables and Figures for Mysid: UNT Lindane Data

27 

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

Chemical: Lindane, Generation: F0 

Parameters Normality 

Test1

Homogeneous of 

Variance Test2

Monotonicity 

Test3

Concentration 

Trend Tests4

Overall 

Concentration 

Effect5

Pairwise 

Comparisons to 

Control6,7

Reproduction 

Survival Rate (%) 

(day 7)

Non-Normal NA Non-Monotonic NA No NA 

Reproduction 

Survival Rate (%) 

(day 14)

Normal Homogeneous Non-Monotonic NA No NA 

Reproduction 

Survival Rate (%) 

(Termination)

Normal Homogeneous Monotonic 

Decreasing trend for 

the highest 

concentration group 

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 

1. Shapiro-Wilk test for normality. 

2. Levene Test for homogeneous of variances. 

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

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

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

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

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

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

conducted: 

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

within-treatment variances for normal distributed data 

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

Summary Tables and Figures for Mysid: UNT Lindane Data

28 

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

Chemical: Lindane, Generation: F1 

Parameters 

Normality 

Test1

Homogeneous of 

Variance Test2

Monotonicity 

Test3

Concentration 

Trend Tests4

Overall 

Concentration 

Effect5

Significant 

Pairwise 

Comparisons 

to Control6,7

Reproduction 

Survival Rate (%) 

(day 7)

Normal Homogeneous Non-Monotonic NA Yes Group 6 

Reproduction 

Survival Rate (%) 

(day 14)

Normal Homogeneous Non-Monotonic NA Yes Group 6 

Reproduction 

Survival Rate (%) 

(Termination)

Normal Homogeneous Non-Monotonic NA No NA 

Total 

Survival Rate (%) 

(day 7)

Non-Normal NA Monotonic 

Decreasing trend for 

the highest 

concentration group 

Yes NA 

Total 

Survival Rate 

(%) (day 14)

Normal Homogeneous Non-Monotonic NA Yes Group 6 

1. Shapiro-Wilk test for normality. 

2. Levene Test for homogeneous of variances. 

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

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

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

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

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

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

conducted: 

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

within-treatment variances for normal distributed data 

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

 
DRAFT 

Summary Tables and Figures for Mysid: UNT Lindane Data

29 

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

Chemical: Lindane. 

Parameters 

Normality 

Test1

Homogeneous 

of Variance 

Test2

Parameteric/ 

Nonparametric 

Concentration 

Main Effect 

Generation 

Main Effect 

Concentration 

by Generation 

Interaction 

Groups with 

Significant 

Difference 

between 

Generations3

Reproduction 

Survival Rate 

(%) 

(day 7)

NonNormal NA Nonparametric Yes No Yes Group 6 

Reproduction 

Survival Rate 

(%) 

(day 14)

Normal Homogeneous Parametric Yes No No NA 

Reproduction 

Survival Rate 

(%) 

(Termination)

Normal Homogeneous Parametric Yes No No NA 

Total 

Survival Rate 

(%) 

(day 7)

NonNormal NA Nonparametric No No No NA 

Total 

Survival Rate 

(%) (day 14)

Normal Homogeneous Parametric Yes Yes Yes Group 6 

1. Shapiro-Wilk test for normality. 

2. Levene test for homogeneous of variances. 

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

 
DRAFT 

Summary Tables and Figures for Mysid: UNT Lindane Data

30 

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

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

From Lab UNT. 

Time To Mature (Days)

9.0

9.5

10.0

10.5

11.0

11.5

12.0

12.5

13.0

13.5

14.0

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Group

Generation 0 Generation 1

Number Offspring (brood 1)

3

4

5

6

7

8

9

10

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Group

Generation 0 Generation 1
DRAFT 

Summary Tables and Figures for Mysid: UNT Lindane Data

31 

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

From Lab UNT. 

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

From Lab UNT. 

Number Offspring (brood 2)

3

4

5

6

7

8

9

10

11

12

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Group

Generation 0 Generation 1

Number Offspring (two broods)

8

10

12

14

16

18

20

22

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Group

Generation 0 Generation 1
DRAFT 

Summary Tables and Figures for Mysid: UNT Lindane Data

32 

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

Figure 6. Days to Brood 2 By Generation and Group For Chemical Lindane From Lab UNT. Days to Brood 1

15

16

17

18

19

20

21

22

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Group

Generation 0 Generation 1

Days to Brood 2

19

20

21

22

23

24

25

26

27

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Group

Generation 0 Generation 1
DRAFT 

Summary Tables and Figures for Mysid: UNT Lindane Data

33 

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

From Lab UNT. 

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

Lindane From Lab UNT. 

Days Between Broods 1 and 2

3

4

5

6

7

8

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Group

Generation 0 Generation 1

% of Females Productively Active

60

65

70

75

80

85

90

95

100

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Group

Generation 0 Generation 1
DRAFT 

Summary Tables and Figures for Mysid: UNT Lindane Data

34 

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

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

Lab UNT. 

Sex Ratio

0.0

0.5

1.0

1.5

2.0

2.5

3.0

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Group

Generation 0 Generation 1

Dry Weight (mg)(day 0)

0.01

0.02

0.03

0.04

0.05

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Group

Generation 0 Generation 1
DRAFT 

Summary Tables and Figures for Mysid: UNT Lindane Data

35 

 
DRAFT 

Summary Tables and Figures for Mysid: UNT Lindane Data

36 

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

Lab UNT. 

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

Lab UNT. 

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

0.34

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Group

Generation 0 Generation 1

Dry Weight (mg)(day 14)

0.40

0.45

0.50

0.55

0.60

0.65

0.70

0.75

0.80

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Group

Generation 0 Generation 1
DRAFT 

Summary Tables and Figures for Mysid: UNT Lindane Data

37 

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

From Lab UNT. 

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

UNT. 

Dry Weight (mg)(Termination)

0.80

0.85

0.90

0.95

1.00

1.05

1.10

1.15

1.20

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Group

Generation 0 Generation 1

Length (mm)(day 0)

1.30

1.35

1.40

1.45

1.50

1.55

1.60

1.65

1.70

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Group

Generation 0 Generation 1
DRAFT 

Summary Tables and Figures for Mysid: UNT Lindane Data

38 

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

UNT. 

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

UNT. 

Length (mm)(day 7)

2.5

3.0

3.5

4.0

4.5

5.0

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Group

Generation 0 Generation 1

Length (mm)(day 14)

4.0

4.5

5.0

5.5

6.0

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Group

Generation 0 Generation 1
DRAFT 

Summary Tables and Figures for Mysid: UNT Lindane Data

39 
DRAFT 

Summary Tables and Figures for Mysid: UNT Lindane Data

40 

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

Lab UNT. 

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

Lindane From Lab UNT. 

Length (mm)(Termination)

5.8

6.0

6.2

6.4

6.6

6.8

7.0

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Group

Generation 0 Generation 1

Reproduction Survival Rate (%)(day 7)

60

70

80

90

100

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Group

Generation 0 Generation 1
DRAFT 

Summary Tables and Figures for Mysid: UNT Lindane Data

41 

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

Lindane From Lab UNT. 

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

Chemical Lindane From Lab UNT. 

Reproduction Survival Rate (%)(day 14)

30

40

50

60

70

80

90

100

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Group

Generation 0 Generation 1

Reproduction Survival Rate (%)(Termination)

30

40

50

60

70

80

90

100

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Group

Generation 0 Generation 1
DRAFT 

Summary Tables and Figures for Mysid: UNT Lindane Data

42 
DRAFT 

Summary Tables and Figures for Mysid: UNT Lindane Data

43 

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

From Lab UNT. 

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

From Lab UNT. 

Total Survival Rate (%)(day 7)

60

65

70

75

80

85

90

95

100

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Group

Generation 0 Generation 1

Total Survival Rate (%)(day 14)

30

40

50

60

70

80

90

100

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Control 0.032

ppb

0.08

ppb

0.2

ppb

0.5

ppb

1.25

ppb

Group

Generation 0 Generation 1