DATA EVALUATION RECORD 

FISH LIFE-CYCLE TOXICITY TEST 

§72-5 

I. CHEMICAL: Ethoprop PC Code No.: 041 101 

2. TEST MATERIAL: Ethoprophos Technical Purity: 94.8% (w:w) 

3. CITATION: 

Author: Dionne, E. 

Title: Ethoprop - Chroni c Toxi city to the Sheepshead Minnow 

(Cyprinodon variegatus) During a Full Life-Cycle 

Exposure. 

Study Completion Date: June 29, 2004 

Laboratories: Springborn Smithers Laboratories 

790 Main Street 

Wareham, MA 0257 1-1075 

Sponsor: Bayer CropScience 

17745 South Metcalf Avenue 

Stilwell, KS 66085 

Laboratory Report ID: 13798.6 154 

~ MRlD No.: 463 15401 

DP Barcode: D359201, D306508 

4. REVIEWED BY: Christie E. Padova, Staff Scientist, Dynamac Corporation 

Signature: Date: 11 /23/04 

APPROVED BY: Teri Myers, Ph.D., Staff Scientist, Dynamac Corporation 

Signature: 

5. APPROVED BY: ;:: p:ing~PP~B-V 

Signature: j V'J'~ 

Date: 1217/04 

Date: 4-30-09 
DATA EVALUATION RECORD

FISH LIFE-CYCLE TOXICITY TEST

§72-5

1. CHEMICAL: Ethoprop PC Code No.: 041101

2. TEST MATERIAL: Ethoprophos Technical Purity: 94.8% (w:w)

3. CITATION:

Author: Dionne, E.

Title: Ethoprop - Chronic Toxicity to the Sheepshead Minnow 

(Cyprinodon variegatus) During a Full Life-Cycle 

Exposure.

Study Completion Date: June 29, 2004

Laboratories: Springborn Smithers Laboratories

790 Main Street

Wareham, MA 02571-1075

Sponsor: Bayer CropScience

17745 South Metcalf Avenue

Stilwell, KS 66085 

Laboratory Report ID: 13798.6154

MRID No.: 46315401

DP Barcode: D359201, D306508

4. REVIEWED BY: Christie E. Padova, Staff Scientist, Dynamac Corporation

Signature: Date: 11/23/04

 APPROVED BY: Teri Myers, Ph.D., Staff Scientist, Dynamac Corporation

Signature: Date: 12/7/04

5. APPROVED BY: Keith Sappington, OPP/EFED/ERB-V

Signature: Date: 4-30-09
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6. STUDY PARAMETERS:

Scientific Name of Test Organism: Cyprinodon variegatus

Age of Test Organism: Embryos, >24 to <48 hours old (F0 generation)

Definitive Test Duration: 112 Days (Approx. 3.5 months)

Study Method: Flow-through

Type of Concentrations: Mean-measured

7. CONCLUSIONS:

The 3.5-month chronic toxicity of Ethoprophos Technical (ethoprop) to the full life stage 

of Sheepshead minnow (Cyprinodon variegatus) was studied under flow-through 

conditions. Fertilized eggs (280 embryos/treatment, >24 to <48 hours old) were exposed to 

the test material at nominal concentrations of 0 (negative control), 1.0, 2.0, 4.0, 8.0, and 16 

ppb a.i. Mean-measured concentrations were <0.22 (control), 1.2, 2.0, 3.7, 7.4, and 15 ppb 

a.i., respectively, and were generally stable during the study. 

Following the completion of hatching on Day 6, larvae were reduced to 100 per treatment 

level. At 4-weeks post-hatch, the juveniles were again reduced to 50 per treatment level. 

Spawning was initiated 8 weeks following hatching: four groups of two male and five 

female per test level were assigned to spawning aquaria, and hatchability trials and early 

life stage studies were performed for the F1 generation. Following hatching, the F1

generation was maintained for 4 weeks. The F0 portion of test was terminated 16 weeks 

after hatching (Day 118).

This study is classified as SUPPLEMENTAL because of several protocol deviations 

involving measurement of embryo viability and poor statistical power for the F1 generation 

endpoints. 

Results Synopsis:

NOAEC: 2.0

LOAEC: 3.7

Endpoint(s) Affected: growth (weight) of 4-week old F1 minnows (32% reduction at 

LOAEC)

8. ADEQUACY OF THE STUDY:

A. Classification: SUPPLEMENTAL
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B. Rationale: 

C. Repairability: This study is not considered upgradable. 

9. GUIDELINE DEVIATIONS:

1. The temperature ranged from 24-27°C throughout the study, which is less than the 

recommended temperature of 30°C for this species. 

2. It was not reported if aeration of the test aquaria was necessary during the study.

3. F1-generation fish were maintained for only 4 weeks, instead of the required 8 

weeks.

4. The reviewer-calculated high-low ratio of mean-measured concentrations was 1.7 

for the nominal 2.0 ppb treatment level, exceeding the 1.5 limit.

5. This study design only included 2 true replicates. Although consistent with the 

minimum number of replicates recommended in the study guideline, the small 

number of replicates may have compromised the statistical power of the test, 

particularly for F1 generation endpoints (see Table 1 below). Since the F1 

endpoints are only addressed by full life cycle studies, these endpoints are of 

particular importance in the study evaluation. For ethoprop, however, the general 

lack of a dose-response was observed for all but one F1 generation endpoint (i.e., 

modest % reductions in mean responses in the treatments relative to controls). 

Thus, for those F1 endpoints where a dose-response was not observed, the poor 

statistical power of the F1 endpoint comparisons is moot. The one exception is 

growth of F1 generation fish measured at 4 weeks post hatch. For this endpoint, a 

32% reduction in mean wet weight of 4-week post hatch minnows relative to 

negative controls occurred at 3.7, 7.4, 15 ppb but these were not found to be 

statistically significant. Reductions in length were of up to 11% were also 

observed in these treatments relative to controls. Since the variance in control 

treatments for these endpoints is very small, the lack of statistical power appears 

to be due to the small number of replicates (2) used in the study. Rather than 

invalidating the study, this reviewer is re-interpreting the study to consider the 

32% reduction in wet weight of 4-week post-hatch F1 minnows at 3.7, 7.4, and 15 

ppb to be biologically significant. Therefore, the NOAEC from this study is 

now interpreted to be 2.0 ppb and the LOAEC is interpreted to be 3.7 ppb, 

based on mean measured concentrations.
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6. Embryo viability measurements were based on a single subsample of 100 

embryos and appears to have introduced bias in the % hatching success 

calculation (see Figure 1 below). However, control % hatching success still 

meets the guideline recommended 75% minimum for sheepshead minnow, when 

Day 1 embryo viability measurements are used instead of the Day 0 subsample. 

Table 1. Minimum Significant Differences (MSD) based on Dunnetts Statistical 

Comparisons from the Ethoprop Fish Full Life Cycle Study (MRID MRID 46315401)

MSD (%) Endpoint Type Generation Comments

18.7 % hatch_F0 hatch F0 Reasonable MSD observed

17.4 % viability surv F0 Reasonable MSD observed

2.9 length_d28 growth F0 Reasonable MSD observed

11.6 weight_d28 growth F0 Reasonable MSD observed

52 % survival_F0 surv F0 Reasonable MSD observed

4.9 length_d28_male growth F0 Reasonable MSD observed

7.6 length_d28_fem growth F0 Reasonable MSD observed

9.4 length_fem growth F0 Reasonable MSD observed

16.6 weight_male growth F0 Reasonable MSD observed

52.9 # eggs_F1 repro F1

17% reduction at 15 ppb, no 

consistent dose-response < 15 ppb (1)

48.3 # eggs/fem_F1 repro F1

16% reduction at 15 ppb, no 

consistent dose-response < 15 ppb(1)

35.4 % hatch_F1 repro F1

25% reduction @ 15 ppb, no 

consistent dose-response < 15 ppb

27.2 length_F1 growth F1

12% reduction @ 15 ppb, 10% @ 7.4 

& 3.7 ppb

62.6 # hatch_d7 hatch F1

2% reduction @ 15 ppb, no

consistent dose-response

>32 weight_F1 growth F1

32% reduction in F1 weight @ 3.7, 

7.4 and 15 ppb (Kruskal Walis)

(1) a cursory analysis of data for this endpoint from 5 fish full life cycle studies suggest that it is typically 

subject to high variability in controls
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Figure 1. Comparison of Corrected Sheepshead Minnow % Hatch: 

Mean Day 1 vs. Day 0 Subsample Embryo Viability 

Relative Difference Between MEAN D0 and D1-Corrected 

Percent Hatch for Sheepshead Minnow FLC

11%

2% 5%

14% 11% 12%

7%

0%

20%

40%

1 2 3 4 5 6 7

Compound 

% Difference Rel. to Day 1

SHM % Hatch Correction:

Day 0 Subsample vs. D1 Viability

60

70

80

90

100

110

120

60 80 100 120

Day 0 Subsample-Corrected

Day 1 ViabilityCorrected

Data

1:1 line

10. SUBMISSION PURPOSE: This study was submitted to provide data on the toxicity of 

ethoprop to the full life-cycle of sheepshead minnows for the purposes of chemical reregistration.

11. MATERIALS AND METHODS:

A. Test Organisms
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Guideline Criteria Reported Information

Species

Prefer Sheepshead minnow (Cyprinodon 

variegatus) or Fathead minnow 

(Pimephales promelas).

Sheepshead minnow (Cyprinodon 

variegatus)

Source and Acclimation Embryos were obtained from Aquatic 

Biosystems, Fort Collins, CO. The 

embryos were held in natural filtered 

seawater at 22°C for approx. 1 hour, then 

at 25°C until assignment. 

Age at beginning of test

Embryos, 2 to 24 hours old Embryos, >24 to <48 hours old

Feeding

Fish should be fed at least twice daily and 

should not be fed for at least 24 hours 

prior to test termination.

F0 and F1 larvae were fed live brine 

shrimp nauplii three times daily during the 

first 28 days post-hatch. After 14 days 

post-hatch, small amounts of dry flaked 

food was also offered daily. The juvenile 

and adult fish were fed frozen brine 

shrimp and Zeigler Prime Flakes twice 

daily. Feedings were withheld 24 hours 

before weight determinations.

Embryo Exposure (4 to 5 Days)

Embryos (≤24 hours old) from at least 3 

separate spawns should be randomly 

distributed to embryo cups.

A minimum of 50 embryos (≤24 hrs old) 

per replicate cup, 4 cups per treatment 

should be used.

Parameters measured:

⋅ Survival of embryos 

⋅ Time required to hatch

⋅ Hatching success

Days 0-6

Embryos (> 24 to <48 hours old) were 

randomly assigned into embryo incubation 

cups; the number of origin spawns was not 

reported.

Each cup contained 70 embryos, with two 

cups per replicate and two replicate 

aquaria per treatment level (total of 280 

embryos per treatment).

Parameters measured:

⋅ Hatching success

⋅ Time to hatch

⋅ Survival of fry at 4 weeks post-hatch
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Guideline Criteria Reported Information

⋅ Survival of fry for 4 weeks

Dead and fungused embryos should be 

counted and removed daily.

Mortality was determined daily. Dead 

embryos were removed.

Larval-Juvenile Exposure (From Hatch 

to 8 Weeks)

After hatching, each group of larvae is 

randomly reduced to a minimum of 25 fish 

and released in replicate larval growth 

chambers. The random selection must 

include any fish that are lethargic or 

deformed.

Parameters measured: 

⋅ Fish survival (determined by 

counting the number of live fish in 

each replicate growth chamber 

weekly).

⋅ Total lengths (mm) of all fish at 4 

and 8 weeks after hatching.

Hatch to 8 Weeks Post-Hatch

When hatching was complete (on Day 6), 

25 larvae were impartially selected from 

each cup and transferred to the larval 

growth chambers (two chambers within 

each replicate aquarium, 100 larvae per 

treatment). At 4 weeks post-hatch, 

juvenile fish from the two growth 

chambers were combined and impartially 

reduced to 25 per replicate (50 per 

treatment). 

Parameters measured:

⋅ Survival of fry/juvenile fish at 8 

weeks post-hatch

⋅ Total lengths (mm) of all surviving 

fish at 4 and 8 weeks post-hatch 

(gender-specific at 8 weeks).

⋅ Wet weights (mg) of fish 

discontinued from exposure (at 

thinning) at 4 weeks post-hatch

Juvenile-Adult Exposure (From 8 weeks 

posthatch to the end of the spawning 

phase [32-40 weeks])

At 20-24 weeks after hatching, mature fish 

are placed in a spawning tank of the same 

concentration (4 males and 4 females 

randomly chosen and assigned). The 

spawning tank is divided into 4 individual 

spawning chambers with appropriate 

spawning substrates.

The substrates are examined daily and 

8 to 16 Weeks Post-Hatch

Two spawning groups (2 male and 5 

female per group) were established for 

each replicate aquarium. The first group 

was established on Day 61 (55 days posthatch), and the second 14 days following 

the first. Females killed by male 

aggression were not replaced; however, 

males were replaced in order to maximize 

egg fertilization success. 

The spawning substrates are examined 

daily and embryos removed, counted, and 
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Guideline Criteria Reported Information

embryos removed, counted, and recorded 

separately for each pair.

For fathead minnow, adult exposure 

should be terminated when no spawning 

occurs for one week. For sheepshead 

minnow, testing should be terminated after 

spawning is observed for 2 weeks.

examined for fertility. 

Adult exposure was terminated on Day 

118 (112 days post-hatch). 

Parameters measured:

⋅ Survival of adult fish

⋅ Fecundity (eggs/female/day)

⋅ Total lengths (mm) and wet weights 

(g) of all surviving fish at Day 118 

(gender-specific)

Second Generation Embryo Exposure

(4 to 5 days)

50 embryos from each conc. level are 

randomly selected and transferred to 

incubation cups for hatch. Use the same 

test procedures as those for parental 

generation.

Embryos not selected are discarded.

F1 Embryo Exposure

50 embryos from spawns of ≥50 eggs were 

incubated in each incubation cup as 

previously described (200 embryos per 

treatment level). 

Second Generation Larval-Juvenile 

Exposure (From Hatch to 4-8 weeks)

After hatching, 25 larvae are released in 

each growth chambers (2 chambers per 

treatment).

Each group of 2nd generation fish is 

terminated 8 weeks after hatching.

Fish are blotted, weighed, and measured 

before being discarded.

F1 Larval-Juvenile Exposure

Groups of 25 newly-hatched larvae were 

randomly released into each larval growth 

chamber (100 larvae per treatment level). 

Each group of F1-generation fish was 

terminated 28 days after hatching.

Fish were weighed (wet) and measured for 

total length.

Comments: 850.1400 guideline requirements specify that the hatching success of the controls for 

sheepshead minnows should be >75%; however, the mean hatching success of the controls in this 

study (57%; page 27 of report) does not meet guideline requirements. A sub-sample of 100 

embryos was examined for viability, and based on visible developmental stages, it was estimated 

that 68% of the eggs were viable (p. 18). Embryo hatching success values were adjusted by the 
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study authors using this estimated percent viability. 

B. Test System

Guideline Criteria Reported Information

Test Water

Sheepshead Minnow

1. Natural seawater (sterilized and 

filtered) or a commercial mixture.

2. Natural seawater with a salinity of 

>15 parts per thousand (weekly range 

of salinity <6% and monthly pH range 

<0.8 pH units).

Fathead Minnow

1. Reconstituted water or water from 

unpolluted well or spring (sterilized 

and tested for pollutants).

2. Hardness of 40 to 48 mg/L as CaCO3

and pH of 7.2 to 7.6.

1. Natural filtered seawater collected 

from Cape Cod Canal, Bourne, MA. 

2. Salinity of 31-33 ‰ and pH 7.6-8.0.

N/A

Test Temperature

Sheepshead: 30°C.

Fathead: 25°C and should not remain 

outside the range of 24 to 26°C for more 

than 48 hours.

24-27°C

N/A

Photo-period

16-hour light/8-hour dark.

Light intensity of 10-100 lumens at water 

surface.

16-hour light/8-hour dark cycle

Light intensity of 52-116 foot-candles 

(560-1200 lux)

Dosing Apparatus

1. Intermittent flow proportional 

diluters or continuous flow serial 

diluters.

2. A minimum of 5 toxicant 

1. Intermittent-flow proportional diluter.

2. Five toxicant concentrations with a 

dilution factor of 0.5.
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Guideline Criteria Reported Information

concentrations with a dilution factor 

<0.5.

3. One control should be used.

3. A dilution water (negative) control was 

used.

Toxicant Mixing

1. Mixing chamber recommended but 

not required.

2. Test solution completely mixed 

before introduction into the test system 

(aeration should not be used for 

mixing).

3. Flow splitting accuracy must be 

within 10% and periodically checked.

1. The diluter system incorporated a 

mixing chamber.

2. Yes

3. The flow-splitting accuracy was <4% 

(p. 19).

Exposure System/Test Vessels

Exposure tanks should be all glass or 

glass with a plastic or stainless steel frame 

(30.5 x 30.5 x 91.4 cm or 30.5 x 30.5 x 61 

cm for fathead, and 45 x 90 x 26 cm for 

sheepshead).

Larval chambers should have glass 

bottoms and drains that allow water to be 

drawn down to 3 cm.

Test water depth in adult tanks and larval 

chambers should be a minimum of 15 cm.

Glass exposure aquaria (60 x 30 x 30 cm) 

were used, with a fill volume of 27 L and 

depth of 15 cm. 

The larval growth chambers were 

constructed of glass and Nitex screening 

and measured 30 x 13 x 25 cm; two larval 

growth chambers were positioned within 

each aquarium. 

During spawning, designated aquaria each 

contained two spawning baskets made of 

8.5-mesh nylon screening. Each basket 

measured 30 cm2 (depth of approx. 12 

cm) and was placed over a removable egg 

collection tray. The trays were 

constructed with 3-cm high glass sides 

and 40-mesh nylon screening bottom. 

Embryo and Fry Chambers

120 mL glass jars with bottoms replaced 

with 40 mesh stainless steel or nylon 

screen. Chambers can be oscillated 

The embryo incubation cups were 5 cm 

diameter glass jars with 40-mesh Nitex 

screen bottoms and stainless steel wire 
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Guideline Criteria Reported Information

vertically using rocker arm apparatus (2 

rpm motor) or placed in separate 

chambers with self-starting siphons.

handles. 

Flow Rate

Flow rates to adult tanks or larval 

chambers should provide 90% 

replacement in 8-12 hours, and maintain 

DO at above 75% of saturation. The 

toxicant level cannot drop below 20% 

with fish in the tank.

During the pre-spawning phase, the flow 

rate was 7.9 volume additions per day.

During the spawning phase, the flow rate 

was 7.7 volume additions per day.

Aeration

Dilution water should be aerated to insure 

dissolved oxygen concentrations at or 

near 100% saturation. Test tanks and 

embryo chambers should not be aerated.

Not specified
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C. Chemical System

Guideline Criteria Reported Information

Nominal Concentrations

Minimum of 5 concentrations and a 

control, all replicated; plus solvent control 

if appropriate.

Toxicant conc. must be measured in one 

tank at each toxicant level every week.

0 (negative control), 1.0, 2.0, 4.0, 8.0, and 

16 ppb a.i.

Toxicant concentrations were measured 

weekly from alternating replicate aquaria 

in each test group. 

Other Variables

1. DO must be measured at each conc. 

at least once a week.

2. Test water temp. must be recorded 

continuously.

3. Freshwater: A control and one conc. 

must be analyzed weekly for pH, 

alkalinity, hardness, and conductance.

Natural seawater: must maintain a 

constant salinity and not fluctuate 

more than 6% weekly; monthly pH 

range <0.8 pH units.

1. DO was measured daily in each 

replicate aquarium.

2. Temperature was measured daily in 

each replicate aquarium, and was also 

continuously monitored in one aquarium 

on both levels of the diluter system. 

3. pH and salinity were measured daily in 

each replicate aquarium. The salinity did 

not fluctuate.

Solvents

Should not exceed 0.1 ml/L in a flowthrough system. Acceptable solvents are: 

dimethylformamide, triethylene glycol, 

methanol, acetone, ethanol.

None used.

Comments: None.

12. REPORTED RESULTS:

A. General Results

Guideline Criteria Reported Information
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Guideline Criteria Reported Information

Quality assurance and GLP compliance 

statements were included in the report? Yes

Data Endpoints must include:

⋅ survival of P and F1 embryos, time 

required to hatch, and hatching 

success;

⋅ survival and total length of P fish at 4 

and 8 weeks after hatching;

⋅ weights and lengths of F1 fish at 8 

weeks;

⋅ incidence of pathological or 

histological effects; and

⋅ observations of other effects or 

clinical signs.

Data Endpoints included:

⋅ survival of F0 and F1 embryos, time 

required to hatch, and hatching 

success;

⋅ survival and total lengths of F0 fish at 

4 and 8 weeks after hatching; 

⋅ wet weights of fish discontinued from 

exposure at thinning 4 weeks after 

hatching; 

⋅ survival of F0 fish at 16 weeks after 

hatching (112 days post-hatch; test 

termination); 

⋅ total lengths and weights (gender 

specific) of surviving F0 fish at 16 

weeks after hatching; 

⋅ F0 fecundity (eggs/female/day)

⋅ total lengths and wet weights of F1

fish at 4 weeks after hatching

⋅ incidence of pathological or 

histological effects;

⋅ observation of other effects or 

clinical signs

Raw data included? Yes
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F0 Results:

Nominal 

Conc.

(ppb a.i.)

Mean Measured 

Conc.

(ppb a.i.) (SD)

%

Hatch1

4-Week PostHatch % 

Survival

8-Week PostHatch % 

Survival2

Terminal

% Survival2, 3

Negative 

Control

<0.22 82 98 98 86

1.0 1.2 ± 0.10 86 100 100 84

2.0 2.0 ± 0.25 82 98 98 76

4.0 3.7 ± 0.28 85 99 98 80

8.0 7.4 ± 0.64 90 99 100 82

16 15 ± 1.3 77 100 98 94

Data obtained from Tables 2, p. 34, and Tables 4-6, pp. 36-38. 1 Adjusted for a mean 68% viability of the eggs used to initiate the study.

2 Based on 50 fish/treatment, thinned at 4 weeks post-hatch. 3 16 weeks post-hatch. 

Mean 

Measured 

Conc.

(ppb a.i.)

Mean Total Length (mm) Wet Weight (g)

4 Weeks 

Posthatch

8 Weeks Post-hatch Test Termination 4 Weeks 

Posthatch

Test Termination

_ _ _ _ _ _

Negative 

Control

28.9 45.2 42.5 53.1 47.5 0.387 3.1 2.1

1.2 28.6 44.7 41.6 53.3 46.9 0.382 3.1 1.9

2.0 28.5 45.1 42.0 53.2 47.3 0.382 3.0 2.1

3.7 28.5 45.6 40.8 53.2 46.8 0.382 3.0 2.1

7.4 28.4 45.5 42.4 54.1 47.4 0.373 3.1 2.0

15 28.0* 44.4 41.9 50.9* 46.1 0.371 2.6* 1.9

Data obtained from Tables 4 and 5, pp. 36-37.

* Significantly reduced compared to the control, based on Williams’ Test.
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Mean Measured Conc.

(ppb a.i.)

Total Number of Eggs1 Number of Eggs/

Female/Day

Negative Control 6888 25

1.2 6622 25

2.0 6770 25

3.7 8309 30

7.4 6876 25

15 5656 22

Data obtained from Table 6, p. 38.

1 Values represents mean of two replicates.

Toxicity Observations: A slight (3%), but statistically-significant reduction in mean total 

length of larval fish was observed at the 15 ppb a.i. group compared to the control (28.0 

versus 28.9 mm) at 4 weeks post-hatch (Table 4, p. 36). No differences in lengths were 

observed at the 8-week measurement; however, at study termination, the mean total length 

and wet weight of males at the 15 ppb a.i. level (50.9 mm and 2.6 g) were statistically less 

than the controls (53.1 mm and 3.1 g). No other treatment-related effects were observed on 

any F0 parameter. All embryo groups, at all exposure levels and the control, hatched in 6 

days (p. 27). No deformities or internal abnormalities were observed among the terminated 

F0 adult fish (p. 28).

F1 Results:

Mean Measured

Concentration

(ppb a.i.)

% Hatch 4 Week

Post-Hatch

% Survival

4 Week

Post-Hatch

Length (mm)

4 Week

Post-Hatch

Wet Weight (g)

Negative Control 68 95 25.1 0.256

1.2 60* 95 21.1 0.160

2.0 71 98 25.0 0.251

3.7 74 93 22.6 0.173

7.4 66 100 22.5 0.175

15 52* 97 21.9 0.173

Data obtained from Table 7, p. 39.

* Significantly reduced compared to the control, based on Fisher’s Exact Test.
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Mean Measured

Concentration

(ppb a.i.)

Percent Hatched

5 Days 6 Days 7 Days 8 Days

Negative Control 0 0 94 6

1.2 0 6 92 2

2.0 4 25 71 0

3.7 0 0 94 6

7.4 0 0 81 19

15 0 8 92 0

Data obtained from Table 8, p. 40.

Toxicity Observations: A statistically-significant reduction in hatching success was 

observed at the 15 ppb a.i. test group compared to the control (52 versus 68%; Table 7, p. 

39). The reduced hatching success at the highest concentration was the result of the 

performance of one spawning group in a total of four maintained during the study (p. 28). 

The mean percent hatching success for the other three groups was 70%. Embryos exposed at 

1.2 ppb a.i. had a statistically-significant reduction in hatching success compared to the 

control (60 versus 68%, respectively); however, this difference was not considered to be 

treatment-related as no concentration response was observed at the three higher levels. No 

other statistical differences were observed on any parameter assessed. 

B. Reported Statistical Results

Data obtained for the F0 generation that were statistically analyzed included hatching 

success; 28-day post-hatch survival, total length and wet weight; 55-day post-hatch survival, 

male total length, and female total length; fecundity (eggs/female/day); 112-day post-hatch 

survival, male total length, male wet weight, female total length, and female wet weight. 

The time to hatch data were empirically estimated. Data obtained for the F1 generation that 

were statistically analyzed included hatching success; time-to-hatch; and 28-day post-hatch 

survival, total length, and wet weight. 

Continuous data (growth and reproduction endpoints) were analyzed for assumptions of 

normality and homogeneity, followed by William’s Test using TOXSTAT (1996) statistical 

software. Binomial data (survival and hatching success endpoints) were analyzed by 

Fisher’s Exact Test using SYSTAT (1999) statistical software. All statistical conclusions 

were made at the 95% level of certainty. 
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The no observed effect concentration (NOEC) is the highest test concentration causing no 

adverse effects. The lowest observed effect concentration (LOEC) is the lowest test 

concentration causing adverse effects. The maximum acceptable toxicant concentration 

(MATC) was calculated as the geometric mean of the NOEC and the LOEC.

Biological Endpoint NOEC

(ppb a.i.)

LOEC

(ppb a.i.)

MATC 

(ppb a.i.)

F0 Generation

Hatching success 15 >15 >15

Time to hatch 15 >15 >15

4-week survival 15 >15 >15

4-week length 7.4 15 11

4-week wet weight 15 >15 >15

8-week survival 15 >15 >15

8-week length, male 15 >15 >15

8-week length, female 15 >15 >15

16-week survival 15 >15 >15

16-week length, male 7.4 15 11

16-week length, female 15 >15 >15

16-week wet weight, male 7.4 15 11

16-week wet weight, female 15 >15 >15

Fecundity (eggs/female/day) 15 >15 >15

F1 Generation

Hatching success 7.4 15 11

Time to hatch 15 >15 >15

4-week survival 15 >15 >15

4-week length 15 >15 >15
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Biological Endpoint NOEC

(ppb a.i.)

LOEC

(ppb a.i.)

MATC 

(ppb a.i.)

4-week weight 15 >15 >15

8-week survival Not 

determined

Not 

determined

Not 

determined

8-week length Not 

determined

Not 

determined

Not 

determined

8-week weight Not 

determined

Not 

determined

Not 

determined

NOEC: 7.4 ppb a.i.

LOEC: 15 ppb a.i.

MATC: 11 ppb a.i. 

Endpoint(s) Affected: F0 4-week post-hatch length; F0 16-week (terminal) post-hatch length 

and wet weight of males; and F1 hatching success (same conclusions)

13. REVIEWER'S STATISTICAL RESULTS: 

Data obtained for the F0 generation that were statistically analyzed included hatching success, 

28-day post-hatch survival, total length and wet weight, 55-day post-hatch survival, male total 

length, and female total length, fecundity (eggs/female/day), 112-day post-hatch survival, male 

total length, male wet weight, female total length, and female wet weight. The time to hatch data 

were also statistically analyzed by comparing the number of groups hatching following treatment 

on days 6, 7, and 8. Data obtained for the F1 generation that were statistically analyzed included 

hatching success, 28-day post-hatch survival, total length, and wet weight. 

All data were analyzed to determine if they satisfied the assumptions of ANOVA (i.e., normality 

and homogeneity of variances). If they did, the NOEC and LOEC were determined using this 

test via TOXSTAT statistical software. If data did not satisfy these assumptions, the NOEC and 

LOEC values were determined using the non-parametric Kruskal-Wallis test. These analyses 

were conducted using TOXSTAT statistical software.
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Biological Endpoint NOEC

(ppb a.i.)

LOEC

(ppb a.i.)

F0 Generation

Hatching success 15 >15

Time to hatch 15 >15

4-week survival 15 >15

4-week length 15 >15

4-week wet weight 15 >15

8-week survival 15 >15

8-week length, male 15 >15

8-week length, female 15 >15

16-week survival 15 >15

16-week length, male 15 >15

16-week length, female 15 >15

16-week wet weight, male 15 >15

16-week wet weight, female 15 >15

Fecundity (eggs/female/day) 15 >15

F1 Generation

Hatching success 15 >15

Time to hatch 15 >15

4-week survival 15 >15

4-week length 15 >15

4-week weight 15 >15

The reviewer’s analyses revealed no significant effects of treatment on any parameter.
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14. REVIEWER'S COMMENTS:

Quality control samples were prepared at each sampling interval and remained with the set 

of exposure solution samples throughout the analytical process; samples were prepared in 

saltwater at a nominal concentrations of 0.500, 4.00, and 20.0 ppb a.i. (p. 24 and Table 3, p. 

35). Recoveries ranged from 82.6-128% (n=51, includes two outliers >120%). 

A method validation study conducted prior to initiation of the definitive test established a 

mean recovery of 91.8 ± 7.47% for ethoprop from filtered seawater (Table 1B, p. 91). The 

LOQ was 0.136 ppb a.i.

This study was performed according to U.S. EPA (FIFRA) Good Laboratory Practice 

Standards (40 CFR 160, 1993) with the exception of the collection for the water and food 

contaminant screening analyses. A Quality Assurance Statement was provided. 

The reviewer’s conclusions differed from the study author’s, in that the reviewer’s analyses 

revealed no significant effects of treatment on any endpoint, while the study author’s 

analyses revealed significant reductions in F0 4-week post-hatch length, F0 16-week 

(terminal) post-hatch length and wet weight of males, and F1 hatching success. Differences 

in these conclusions are attributed to the different statistical methods that these results are 

based on. In the reviewer’s analysis, ANOVA did not reveal any significant differences 

among the control and treatment groups, so post-hoc multiple comparison tests (i.e., 

William’s test) were not warranted. 

However, the number of true replicates used in the study is small (2) but does comply with 

the minimum specifications in the supporting documents of the 850.1500 guideline (Hansen 

et al., 1978). Regardless of the number of replicates used in a test, the primary issue of 

concern is the extent to which statistically significant differences can be detected relative to 

the control. An analysis of the minimum significant difference (MSD) reported for the 

ethoprop study indicates that the MSD is very large for some endpoints particularly from the 

F1 generation fish (see Table 1 above). Since the F1 endpoints are only addressed by full 

life cycle studies, these endpoints are of particular importance in the study evaluation. For 

ethoprop, however, the general lack of a dose-response was observed for all but one F1 

generation endpoint (i.e., modest % reductions in mean responses in the treatments relative 

to controls). Thus, for those F1 endpoints where a dose-response was not observed, the poor 

statistical power of the F1 endpoint comparisons is moot. The one exception is growth of F1 

generation fish measured at 4 weeks post hatch. For this endpoint, a 32% reduction in 

mean wet weight of 4-week post hatch minnows relative to negative controls occurred 

at 3.7, 7.4, 15 ppb but these were not found to be statistically significant. Reductions in 

length were of up to 11% were also observed in these treatments relative to controls. Since 

the variance in control treatments for these endpoints is very small, the lack of statistical 

power appears to be due to the small number of replicates (2) used in the study. Rather than 
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21

invalidating the study, this reviewer is re-interpreting the study to consider the 32% 

reduction in wet weight of 4-week post-hatch F1 minnows at 3.7, 7.4, and 15 ppb to be 

biologically significant. Therefore, the NOAEC from this study is now interpreted to 

be 2.0 ppb and the LOAEC is interpreted to be 3.7 ppb, based on mean measured 

concentrations.

15. REFERENCES:

APHA, AWWA, WPCF. 1995. Standard Methods for the Examination of Water and 

Wastewater. 19th Edition, Washington, DC, 2-23-25.

ASTM. 2002. Standard practice for conducting acute toxicity tests with fishes, 

macroinvertebrates and amphibians. Standard E729-96. American Society for Testing and 

Materials, 100 Barr Harbor Drive, West Conshohocken, PA 19428.

Hansen, D.L., et al. 1978. Life-cycle toxicity test using sheepshead minnow (Cyprinodon 

variegatus). In: Bioassay Procedures for the Ocean Disposal Permit Program. EPA-600/9-

78-010, March 1978.

Macek, K.J. and B.H. Sleight, III. 1977. Utility of toxicity tests with embryo and fry of fish in 

evaluating hazards associated with chronic toxicity of chemicals to fishes. Symposium 

Proceedings, ASTM, Memphis, Tennessee, October, 1976: 137-146.

McKim, J.N. and D.A. Benoit. 1971. Effect of long-term exposure to copper on survival, 

reproduction and growth of brook trout (Salvelinus fontinalis). J. Fish. Res. Bd. Can. 28: 

655-662.

McKim, J.M. 1977. Evaluation of tests with early-life-stage of fish for predicting long-term 

toxicity. J. Fish. Res. Board Can. 34: 1148-1154.

Mount, D.I. and W.A. Brungs. 1967. A simplified dosing apparatus for fish toxicological 

studies. Water Res. 1:20-29.

Rexrode, M. and T.M. Armitage. 1986. Standard Evaluation Procedure; Fish Life-Cycle 

Toxicity Tests. Hazard Evaluation Division, Office of Pesticide Programs. Washington, 

D.C. 20460. EPA 540/9-86-137.

Sprague, J.B. 1969. Measurement of pollutant toxicity to fish. 1. Bioassay methods for acute 

toxicity. Water Res. 1: 20-29.

SYSTAT®. 1999. SPSS, Inc.
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22

U.S. EPA. 1989. Federal Insecticide, Fungicide and Rodenticide Act (FIFRA); Good Laboratory 

Practice Standards; Final Rule (40 CFR, Part 160).

U.S. Environmental Protection Agency. 1996. Ecological Effects Test Guidelines, OPPTS 

850.1400: Fish Early-Life Stage Toxicity Test, (Public Draft), Environmental Protection 

Agency, Office of Pesticide Programs. Washington, D.C.

West, Inc., and D.D. Gulley. 1996. Toxstat, Release 3.5. West, Inc., Cheyenne, WY.
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APPENDIX I. OUTPUT OF REVIEWER’S STATISTICAL VERIFICATION:

hatching success

File: 5401hs Transform: NO TRANSFORMATION

 ANOVA TABLE

------------------------------------------------------------------------------

SOURCE DF SS MS F

------------------------------------------------------------------------------

Between 5 80.667 16.133 1.152

Within (Error) 6 84.000 14.000

------------------------------------------------------------------------------

Total 11 164.667

------------------------------------------------------------------------------

 Critical F value = 4.39 (0.05,5,6)

 Since F < Critical F FAIL TO REJECT Ho:All groups equal

hatching success

File: 5401hs Transform: NO TRANSFORMATION

 DUNNETTS TEST - TABLE 1 OF 2 Ho:Control<Treatment

----------------------------------------------------------------------------

 TRANSFORMED MEAN CALCULATED IN 

GROUP IDENTIFICATION MEAN ORIGINAL UNITS T STAT SIG

----- -------------------- ----------- ------------------ ------ ---

 1 control 56.500 56.500

 2 1.2 59.000 59.000 -0.668

 3 2.0 56.500 56.500 0.000

 4 3.7 57.500 57.500 -0.267

 5 7.4 61.500 61.500 -1.336

 6 15 53.000 53.000 0.935

----------------------------------------------------------------------------

Dunnett table value = 2.83 (1 Tailed Value, P=0.05, df=6,5)
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hatching success

File: 5401hs Transform: NO TRANSFORMATION

 DUNNETTS TEST - TABLE 2 OF 2 Ho:Control<Treatment

----------------------------------------------------------------------------

 NUM OF Minimum Sig Diff % of DIFFERENCE

GROUP IDENTIFICATION REPS (IN ORIG. UNITS) CONTROL FROM CONTROL

----- -------------------- ------- ---------------- ------- ------------

 1 control 2

 2 1.2 2 10.589 18.7 -2.500

 3 2.0 2 10.589 18.7 0.000

 4 3.7 2 10.589 18.7 -1.000

 5 7.4 2 10.589 18.7 -5.000

 6 15 2 10.589 18.7 3.500

------------------------------------------------------------------------------

hatching success

File: 5401hs Transform: NO TRANSFORMATION

 WILLIAMS TEST (Isotonic regression model) TABLE 1 OF 2

----------------------------------------------------------------------------

GROUP ORIGINAL TRANSFORMED ISOTONIZED 

 IDENTIFICATION N MEAN MEAN MEAN 

------ -------------------- --- ----------- ----------- -----------

 1 control 2 56.500 56.500 58.200

 2 1.2 2 59.000 59.000 58.200

 3 2.0 2 56.500 56.500 58.200

 4 3.7 2 57.500 57.500 58.200

 5 7.4 2 61.500 61.500 58.200

 6 15 2 53.000 53.000 53.000

----------------------------------------------------------------------------
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25

hatching success

File: 5401hs Transform: NO TRANSFORMATION

 WILLIAMS TEST (Isotonic regression model) TABLE 2 OF 2

----------------------------------------------------------------------------

 ISOTONIZED CALC. SIG TABLE DEGREES OF

 IDENTIFICATION MEAN WILLIAMS P=.05 WILLIAMS FREEDOM

-------------------- ----------- ----------- ----- ----------- -------------

 control 58.200

 1.2 58.200 0.454 1.94 k= 1, v= 6

 2.0 58.200 0.454 2.06 k= 2, v= 6

 3.7 58.200 0.454 2.10 k= 3, v= 6

 7.4 58.200 0.454 2.12 k= 4, v= 6

 15 53.000 0.935 2.13 k= 5, v= 6

----------------------------------------------------------------------------

s = 3.742

Note: df used for table values are approximate when v > 20.

hatching success (% viability)

File: 5401hsv Transform: NO TRANSFORMATION

 ANOVA TABLE

------------------------------------------------------------------------------

SOURCE DF SS MS F

------------------------------------------------------------------------------

Between 5 194.667 38.933 1.557

Within (Error) 6 150.000 25.000

------------------------------------------------------------------------------

Total 11 344.667

------------------------------------------------------------------------------

 Critical F value = 4.39 (0.05,5,6)

 Since F < Critical F FAIL TO REJECT Ho:All groups equal
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26

hatching success (% viability)

File: 5401hsv Transform: NO TRANSFORMATION

 DUNNETTS TEST - TABLE 1 OF 2 Ho:Control<Treatment

----------------------------------------------------------------------------

 TRANSFORMED MEAN CALCULATED IN 

GROUP IDENTIFICATION MEAN ORIGINAL UNITS T STAT SIG

----- -------------------- ----------- ------------------ ------ ---

 1 control 81.500 81.500

 2 1.2 85.500 85.500 -0.800

 3 2.0 81.500 81.500 0.000

 4 3.7 84.500 84.500 -0.600

 5 7.4 90.000 90.000 -1.700

 6 15 77.000 77.000 0.900

----------------------------------------------------------------------------

Dunnett table value = 2.83 (1 Tailed Value, P=0.05, df=6,5)

hatching success (% viability)

File: 5401hsv Transform: NO TRANSFORMATION

 DUNNETTS TEST - TABLE 2 OF 2 Ho:Control<Treatment

----------------------------------------------------------------------------

 NUM OF Minimum Sig Diff % of DIFFERENCE

GROUP IDENTIFICATION REPS (IN ORIG. UNITS) CONTROL FROM CONTROL

----- -------------------- ------- ---------------- ------- ------------

 1 control 2

 2 1.2 2 14.150 17.4 -4.000

 3 2.0 2 14.150 17.4 0.000

 4 3.7 2 14.150 17.4 -3.000

 5 7.4 2 14.150 17.4 -8.500

 6 15 2 14.150 17.4 4.500

------------------------------------------------------------------------------

hatching success (% viability)
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27

File: 5401hsv Transform: NO TRANSFORMATION

 WILLIAMS TEST (Isotonic regression model) TABLE 1 OF 2

----------------------------------------------------------------------------

GROUP ORIGINAL TRANSFORMED ISOTONIZED 

 IDENTIFICATION N MEAN MEAN MEAN 

------ -------------------- --- ----------- ----------- -----------

 1 control 2 81.500 81.500 84.600

 2 1.2 2 85.500 85.500 84.600

 3 2.0 2 81.500 81.500 84.600

 4 3.7 2 84.500 84.500 84.600

 5 7.4 2 90.000 90.000 84.600

 6 15 2 77.000 77.000 77.000

----------------------------------------------------------------------------

hatching success (% viability)

File: 5401hsv Transform: NO TRANSFORMATION

 WILLIAMS TEST (Isotonic regression model) TABLE 2 OF 2

----------------------------------------------------------------------------

 ISOTONIZED CALC. SIG TABLE DEGREES OF

 IDENTIFICATION MEAN WILLIAMS P=.05 WILLIAMS FREEDOM

-------------------- ----------- ----------- ----- ----------- -------------

 control 84.600

 1.2 84.600 0.620 1.94 k= 1, v= 6

 2.0 84.600 0.620 2.06 k= 2, v= 6

 3.7 84.600 0.620 2.10 k= 3, v= 6

 7.4 84.600 0.620 2.12 k= 4, v= 6

 15 77.000 0.900 2.13 k= 5, v= 6

----------------------------------------------------------------------------

s = 5.000

Note: df used for table values are approximate when v > 20.

day 28 length

File: 5401l28 Transform: NO TRANSFORMATION

 ANOVA TABLE
DP Barcode: D306508 MRID No: 46315401

28

------------------------------------------------------------------------------

SOURCE DF SS MS F

------------------------------------------------------------------------------

Between 5 0.998 0.200 2.198

Within (Error) 6 0.545 0.091

------------------------------------------------------------------------------

Total 11 1.543

------------------------------------------------------------------------------

 Critical F value = 4.39 (0.05,5,6)

 Since F < Critical F FAIL TO REJECT Ho:All groups equal

day 28 length

File: 5401l28 Transform: NO TRANSFORMATION

 DUNNETTS TEST - TABLE 1 OF 2 Ho:Control<Treatment

----------------------------------------------------------------------------

 TRANSFORMED MEAN CALCULATED IN 

GROUP IDENTIFICATION MEAN ORIGINAL UNITS T STAT SIG

----- -------------------- ----------- ------------------ ------ ---

 1 control 28.950 28.950

 2 1.2 28.650 28.650 0.994

 3 2.0 28.450 28.450 1.657

 4 3.7 28.450 28.450 1.657

 5 7.4 28.350 28.350 1.989

 6 15 28.000 28.000 3.149 *

----------------------------------------------------------------------------

Dunnett table value = 2.83 (1 Tailed Value, P=0.05, df=6,5)

day 28 length

File: 5401l28 Transform: NO TRANSFORMATION
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29

 DUNNETTS TEST - TABLE 2 OF 2 Ho:Control<Treatment

----------------------------------------------------------------------------

 NUM OF Minimum Sig Diff % of DIFFERENCE

GROUP IDENTIFICATION REPS (IN ORIG. UNITS) CONTROL FROM CONTROL

----- -------------------- ------- ---------------- ------- ------------

 1 control 2

 2 1.2 2 0.854 2.9 0.300

 3 2.0 2 0.854 2.9 0.500

 4 3.7 2 0.854 2.9 0.500

 5 7.4 2 0.854 2.9 0.600

 6 15 2 0.854 2.9 0.950

------------------------------------------------------------------------------

day 28 length

File: 5401l28 Transform: NO TRANSFORMATION

 WILLIAMS TEST (Isotonic regression model) TABLE 1 OF 2

----------------------------------------------------------------------------

GROUP ORIGINAL TRANSFORMED ISOTONIZED 

 IDENTIFICATION N MEAN MEAN MEAN 

------ -------------------- --- ----------- ----------- -----------

 1 control 2 28.950 28.950 28.950

 2 1.2 2 28.650 28.650 28.650

 3 2.0 2 28.450 28.450 28.450

 4 3.7 2 28.450 28.450 28.450

 5 7.4 2 28.350 28.350 28.350

 6 15 2 28.000 28.000 28.000

----------------------------------------------------------------------------

day 28 length

File: 5401l28 Transform: NO TRANSFORMATION

 WILLIAMS TEST (Isotonic regression model) TABLE 2 OF 2

----------------------------------------------------------------------------
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30

 ISOTONIZED CALC. SIG TABLE DEGREES OF

 IDENTIFICATION MEAN WILLIAMS P=.05 WILLIAMS FREEDOM

-------------------- ----------- ----------- ----- ----------- -------------

 control 28.950

 1.2 28.650 0.995 1.94 k= 1, v= 6

 2.0 28.450 1.659 2.06 k= 2, v= 6

 3.7 28.450 1.659 2.10 k= 3, v= 6

 7.4 28.350 1.991 2.12 k= 4, v= 6

 15 28.000 3.152 * 2.13 k= 5, v= 6

----------------------------------------------------------------------------

s = 0.301

Note: df used for table values are approximate when v > 20.

day 28 weight

File: 5401w28 Transform: NO TRANSFORMATION

 ANOVA TABLE

------------------------------------------------------------------------------

SOURCE DF SS MS F

------------------------------------------------------------------------------

Between 5 387.000 77.400 0.308

Within (Error) 6 1510.000 251.667

------------------------------------------------------------------------------

Total 11 1897.000

------------------------------------------------------------------------------

 Critical F value = 4.39 (0.05,5,6)

 Since F < Critical F FAIL TO REJECT Ho:All groups equal

day 28 weight

File: 5401w28 Transform: NO TRANSFORMATION

 DUNNETTS TEST - TABLE 1 OF 2 Ho:Control<Treatment

----------------------------------------------------------------------------
DP Barcode: D306508 MRID No: 46315401

31

 TRANSFORMED MEAN CALCULATED IN 

GROUP IDENTIFICATION MEAN ORIGINAL UNITS T STAT SIG

----- -------------------- ----------- ------------------ ------ ---

 1 control 387.500 387.500

 2 1.2 381.500 381.500 0.378

 3 2.0 382.000 382.000 0.347

 4 3.7 382.000 382.000 0.347

 5 7.4 372.500 372.500 0.946

 6 15 371.500 371.500 1.009

----------------------------------------------------------------------------

Dunnett table value = 2.83 (1 Tailed Value, P=0.05, df=6,5)

day 28 weight

File: 5401w28 Transform: NO TRANSFORMATION

 DUNNETTS TEST - TABLE 2 OF 2 Ho:Control<Treatment

----------------------------------------------------------------------------

 NUM OF Minimum Sig Diff % of DIFFERENCE

GROUP IDENTIFICATION REPS (IN ORIG. UNITS) CONTROL FROM CONTROL

----- -------------------- ------- ---------------- ------- ------------

 1 control 2

 2 1.2 2 44.895 11.6 6.000

 3 2.0 2 44.895 11.6 5.500

 4 3.7 2 44.895 11.6 5.500

 5 7.4 2 44.895 11.6 15.000

 6 15 2 44.895 11.6 16.000

------------------------------------------------------------------------------

day 28 weight

File: 5401w28 Transform: NO TRANSFORMATION

 WILLIAMS TEST (Isotonic regression model) TABLE 1 OF 2

----------------------------------------------------------------------------

GROUP ORIGINAL TRANSFORMED ISOTONIZED 
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32

 IDENTIFICATION N MEAN MEAN MEAN 

------ -------------------- --- ----------- ----------- -----------

 1 control 2 387.500 387.500 387.500

 2 1.2 2 381.500 381.500 381.833

 3 2.0 2 382.000 382.000 381.833

 4 3.7 2 382.000 382.000 381.833

 5 7.4 2 372.500 372.500 372.500

 6 15 2 371.500 371.500 371.500

----------------------------------------------------------------------------

day 28 weight

File: 5401w28 Transform: NO TRANSFORMATION

 WILLIAMS TEST (Isotonic regression model) TABLE 2 OF 2

----------------------------------------------------------------------------

 ISOTONIZED CALC. SIG TABLE DEGREES OF

 IDENTIFICATION MEAN WILLIAMS P=.05 WILLIAMS FREEDOM

-------------------- ----------- ----------- ----- ----------- -------------

 control 387.500

 1.2 381.833 0.357 1.94 k= 1, v= 6

 2.0 381.833 0.357 2.06 k= 2, v= 6

 3.7 381.833 0.357 2.10 k= 3, v= 6

 7.4 372.500 0.946 2.12 k= 4, v= 6

 15 371.500 1.009 2.13 k= 5, v= 6

----------------------------------------------------------------------------

s = 15.864

Note: df used for table values are approximate when v > 20.

day 55 length males

File: 5401l55 Transform: NO TRANSFORM

 KRUSKAL-WALLIS ANOVA BY RANKS - TABLE 1 OF 2

----------------------------------------------------------------------------

 TRANSFORMED MEAN CALCULATED IN RANK 

GROUP IDENTIFICATION MEAN ORIGINAL UNITS SUM 

----- -------------------- ----------- ------------------ -----------
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33

 1 control 45.250 45.250 13.000

 2 1.2 44.750 44.750 10.500

 3 2.0 45.100 45.100 9.500

 4 3.7 45.600 45.600 18.500

 5 7.4 45.500 45.500 21.000

 6 15 44.450 44.450 5.500

----------------------------------------------------------------------------

 Calculated H Value = 6.592 Critical H Value Table = 11.070

 Since Calc H < Crit H FAIL TO REJECT Ho:All groups are equal.

day 55 length males

File: 5401l55 Transform: NO TRANSFORM

DUNNS MULTIPLE COMPARISON - KRUSKAL-WALLIS - TABLE 2 OF 2

----------------------------------------------------------------------------

 GROUP

 TRANSFORMED ORIGINAL 0 0 0 0 0 0 

GROUP IDENTIFICATION MEAN MEAN 6 2 3 1 5 4 

----- --------------- ----------- --------- - - - - - - 

 6 15 44.450 44.450 \

 2 1.2 44.750 44.750 . \

 3 2.0 45.100 45.100 . . \

 1 control 45.250 45.250 . . . \

 5 7.4 45.500 45.500 . . . . \

 4 3.7 45.600 45.600 . . . . . \

----------------------------------------------------------------------------

* = significant difference (p=0.05) . = no significant difference

Table q value (0.05,6) = 2.936 SE = 3.580

day 55 female length

File: 540155f Transform: NO TRANSFORMATION

 ANOVA TABLE

------------------------------------------------------------------------------
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34

SOURCE DF SS MS F

------------------------------------------------------------------------------

Between 5 4.537 0.907 0.696

Within (Error) 6 7.820 1.303

------------------------------------------------------------------------------

Total 11 12.357

------------------------------------------------------------------------------

 Critical F value = 4.39 (0.05,5,6)

 Since F < Critical F FAIL TO REJECT Ho:All groups equal

day 55 female length

File: 540155f Transform: NO TRANSFORMATION

 DUNNETTS TEST - TABLE 1 OF 2 Ho:Control<Treatment

----------------------------------------------------------------------------

 TRANSFORMED MEAN CALCULATED IN 

GROUP IDENTIFICATION MEAN ORIGINAL UNITS T STAT SIG

----- -------------------- ----------- ------------------ ------ ---

 1 control 42.550 42.550

 2 1.2 41.600 41.600 0.832

 3 2.0 42.000 42.000 0.482

 4 3.7 40.750 40.750 1.577

 5 7.4 42.550 42.550 0.000

 6 15 41.850 41.850 0.613

----------------------------------------------------------------------------

Dunnett table value = 2.83 (1 Tailed Value, P=0.05, df=6,5)

day 55 female length

File: 540155f Transform: NO TRANSFORMATION

 DUNNETTS TEST - TABLE 2 OF 2 Ho:Control<Treatment
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35

----------------------------------------------------------------------------

 NUM OF Minimum Sig Diff % of DIFFERENCE

GROUP IDENTIFICATION REPS (IN ORIG. UNITS) CONTROL FROM CONTROL

----- -------------------- ------- ---------------- ------- ------------

 1 control 2

 2 1.2 2 3.230 7.6 0.950

 3 2.0 2 3.230 7.6 0.550

 4 3.7 2 3.230 7.6 1.800

 5 7.4 2 3.230 7.6 0.000

 6 15 2 3.230 7.6 0.700

------------------------------------------------------------------------------

day 55 female length

File: 540155f Transform: NO TRANSFORMATION

 WILLIAMS TEST (Isotonic regression model) TABLE 1 OF 2

----------------------------------------------------------------------------

GROUP ORIGINAL TRANSFORMED ISOTONIZED 

 IDENTIFICATION N MEAN MEAN MEAN 

------ -------------------- --- ----------- ----------- -----------

 1 control 2 42.550 42.550 42.550

 2 1.2 2 41.600 41.600 41.800

 3 2.0 2 42.000 42.000 41.800

 4 3.7 2 40.750 40.750 41.717

 5 7.4 2 42.550 42.550 41.717

 6 15 2 41.850 41.850 41.717

----------------------------------------------------------------------------

day 55 female length

File: 540155f Transform: NO TRANSFORMATION

 WILLIAMS TEST (Isotonic regression model) TABLE 2 OF 2

----------------------------------------------------------------------------

 ISOTONIZED CALC. SIG TABLE DEGREES OF
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36

 IDENTIFICATION MEAN WILLIAMS P=.05 WILLIAMS FREEDOM

-------------------- ----------- ----------- ----- ----------- -------------

 control 42.550

 1.2 41.800 0.657 1.94 k= 1, v= 6

 2.0 41.800 0.657 2.06 k= 2, v= 6

 3.7 41.717 0.730 2.10 k= 3, v= 6

 7.4 41.717 0.730 2.12 k= 4, v= 6

 15 41.717 0.730 2.13 k= 5, v= 6

----------------------------------------------------------------------------

s = 1.142

Note: df used for table values are approximate when v > 20.

F0 survival

File: 5401s Transform: NO TRANSFORMATION

 ANOVA TABLE

------------------------------------------------------------------------------

SOURCE DF SS MS F

------------------------------------------------------------------------------

Between 5 374.667 74.933 0.301

Within (Error) 6 1496.000 249.333

------------------------------------------------------------------------------

Total 11 1870.667

------------------------------------------------------------------------------

 Critical F value = 4.39 (0.05,5,6)

 Since F < Critical F FAIL TO REJECT Ho:All groups equal

F0 survival

File: 5401s Transform: NO TRANSFORMATION

 DUNNETTS TEST - TABLE 1 OF 2 Ho:Control<Treatment

----------------------------------------------------------------------------

 TRANSFORMED MEAN CALCULATED IN 
DP Barcode: D306508 MRID No: 46315401

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GROUP IDENTIFICATION MEAN ORIGINAL UNITS T STAT SIG

----- -------------------- ----------- ------------------ ------ ---

 1 control 86.000 86.000

 2 1.2 84.000 84.000 0.127

 3 2.0 76.000 76.000 0.633

 4 3.7 80.000 80.000 0.380

 5 7.4 82.000 82.000 0.253

 6 15 94.000 94.000 -0.507

----------------------------------------------------------------------------

Dunnett table value = 2.83 (1 Tailed Value, P=0.05, df=6,5)

F0 survival

File: 5401s Transform: NO TRANSFORMATION

 DUNNETTS TEST - TABLE 2 OF 2 Ho:Control<Treatment

----------------------------------------------------------------------------

 NUM OF Minimum Sig Diff % of DIFFERENCE

GROUP IDENTIFICATION REPS (IN ORIG. UNITS) CONTROL FROM CONTROL

----- -------------------- ------- ---------------- ------- ------------

 1 control 2

 2 1.2 2 44.686 52.0 2.000

 3 2.0 2 44.686 52.0 10.000

 4 3.7 2 44.686 52.0 6.000

 5 7.4 2 44.686 52.0 4.000

 6 15 2 44.686 52.0 -8.000

------------------------------------------------------------------------------

F0 survival

File: 5401s Transform: NO TRANSFORMATION

 WILLIAMS TEST (Isotonic regression model) TABLE 1 OF 2

----------------------------------------------------------------------------

GROUP ORIGINAL TRANSFORMED ISOTONIZED 

 IDENTIFICATION N MEAN MEAN MEAN 
DP Barcode: D306508 MRID No: 46315401

38

------ -------------------- --- ----------- ----------- -----------

 1 control 2 86.000 86.000 81.500

 2 1.2 2 84.000 84.000 81.500

 3 2.0 2 76.000 76.000 81.500

 4 3.7 2 80.000 80.000 81.500

 5 7.4 2 82.000 82.000 82.000

 6 15 2 94.000 94.000 94.000

----------------------------------------------------------------------------

F0 survival

File: 5401s Transform: NO TRANSFORMATION

 WILLIAMS TEST (Isotonic regression model) TABLE 2 OF 2

----------------------------------------------------------------------------

 ISOTONIZED CALC. SIG TABLE DEGREES OF

 IDENTIFICATION MEAN WILLIAMS P=.05 WILLIAMS FREEDOM

-------------------- ----------- ----------- ----- ----------- -------------

 control 81.500

 1.2 81.500 0.285 1.94 k= 1, v= 6

 2.0 81.500 0.285 2.06 k= 2, v= 6

 3.7 81.500 0.285 2.10 k= 3, v= 6

 7.4 82.000 0.253 2.12 k= 4, v= 6

 15 94.000 0.507 2.13 k= 5, v= 6

----------------------------------------------------------------------------

s = 15.790

Note: df used for table values are approximate when v > 20.

F0 male length

File: 5401ml Transform: NO TRANSFORMATION

 ANOVA TABLE

------------------------------------------------------------------------------

SOURCE DF SS MS F

------------------------------------------------------------------------------

Between 5 12.867 2.573 3.034
DP Barcode: D306508 MRID No: 46315401

39

Within (Error) 6 5.090 0.848

------------------------------------------------------------------------------

Total 11 17.957

------------------------------------------------------------------------------

 Critical F value = 4.39 (0.05,5,6)

 Since F < Critical F FAIL TO REJECT Ho:All groups equal

F0 male length

File: 5401ml Transform: NO TRANSFORMATION

 DUNNETTS TEST - TABLE 1 OF 2 Ho:Control<Treatment

----------------------------------------------------------------------------

 TRANSFORMED MEAN CALCULATED IN 

GROUP IDENTIFICATION MEAN ORIGINAL UNITS T STAT SIG

----- -------------------- ----------- ------------------ ------ ---

 1 control 53.000 53.000

 2 1.2 53.350 53.350 -0.380

 3 2.0 53.300 53.300 -0.326

 4 3.7 53.400 53.400 -0.434

 5 7.4 54.200 54.200 -1.303

 6 15 50.850 50.850 2.335

----------------------------------------------------------------------------

Dunnett table value = 2.83 (1 Tailed Value, P=0.05, df=6,5)

F0 male length

File: 5401ml Transform: NO TRANSFORMATION

 DUNNETTS TEST - TABLE 2 OF 2 Ho:Control<Treatment

----------------------------------------------------------------------------

 NUM OF Minimum Sig Diff % of DIFFERENCE

GROUP IDENTIFICATION REPS (IN ORIG. UNITS) CONTROL FROM CONTROL

----- -------------------- ------- ---------------- ------- ------------
DP Barcode: D306508 MRID No: 46315401

40

 1 control 2

 2 1.2 2 2.606 4.9 -0.350

 3 2.0 2 2.606 4.9 -0.300

 4 3.7 2 2.606 4.9 -0.400

 5 7.4 2 2.606 4.9 -1.200

 6 15 2 2.606 4.9 2.150

------------------------------------------------------------------------------

F0 male length

File: 5401ml Transform: NO TRANSFORMATION

 WILLIAMS TEST (Isotonic regression model) TABLE 1 OF 2

----------------------------------------------------------------------------

GROUP ORIGINAL TRANSFORMED ISOTONIZED 

 IDENTIFICATION N MEAN MEAN MEAN 

------ -------------------- --- ----------- ----------- -----------

 1 control 2 53.000 53.000 53.450

 2 1.2 2 53.350 53.350 53.450

 3 2.0 2 53.300 53.300 53.450

 4 3.7 2 53.400 53.400 53.450

 5 7.4 2 54.200 54.200 53.450

 6 15 2 50.850 50.850 50.850

----------------------------------------------------------------------------

F0 male length

File: 5401ml Transform: NO TRANSFORMATION

 WILLIAMS TEST (Isotonic regression model) TABLE 2 OF 2

----------------------------------------------------------------------------

 ISOTONIZED CALC. SIG TABLE DEGREES OF

 IDENTIFICATION MEAN WILLIAMS P=.05 WILLIAMS FREEDOM

-------------------- ----------- ----------- ----- ----------- -------------

 control 53.450

 1.2 53.450 0.489 1.94 k= 1, v= 6
DP Barcode: D306508 MRID No: 46315401

41

 2.0 53.450 0.489 2.06 k= 2, v= 6

 3.7 53.450 0.489 2.10 k= 3, v= 6

 7.4 53.450 0.489 2.12 k= 4, v= 6

 15 50.850 2.334 * 2.13 k= 5, v= 6

----------------------------------------------------------------------------

s = 0.921

Note: df used for table values are approximate when v > 20.

F0 female length

File: 5401fl Transform: NO TRANSFORMATION

 ANOVA TABLE

------------------------------------------------------------------------------

SOURCE DF SS MS F

------------------------------------------------------------------------------

Between 5 2.907 0.581 0.236

Within (Error) 6 14.760 2.460

------------------------------------------------------------------------------

Total 11 17.667

------------------------------------------------------------------------------

 Critical F value = 4.39 (0.05,5,6)

 Since F < Critical F FAIL TO REJECT Ho:All groups equal

F0 female length

File: 5401fl Transform: NO TRANSFORMATION

 DUNNETTS TEST - TABLE 1 OF 2 Ho:Control<Treatment

----------------------------------------------------------------------------

 TRANSFORMED MEAN CALCULATED IN 

GROUP IDENTIFICATION MEAN ORIGINAL UNITS T STAT SIG

----- -------------------- ----------- ------------------ ------ ---

 1 control 47.400 47.400

 2 1.2 47.000 47.000 0.255
DP Barcode: D306508 MRID No: 46315401

42

 3 2.0 47.100 47.100 0.191

 4 3.7 46.300 46.300 0.701

 5 7.4 47.300 47.300 0.064

 6 15 46.100 46.100 0.829

----------------------------------------------------------------------------

Dunnett table value = 2.83 (1 Tailed Value, P=0.05, df=6,5)

F0 female length

File: 5401fl Transform: NO TRANSFORMATION

 DUNNETTS TEST - TABLE 2 OF 2 Ho:Control<Treatment

----------------------------------------------------------------------------

 NUM OF Minimum Sig Diff % of DIFFERENCE

GROUP IDENTIFICATION REPS (IN ORIG. UNITS) CONTROL FROM CONTROL

----- -------------------- ------- ---------------- ------- ------------

 1 control 2

 2 1.2 2 4.439 9.4 0.400

 3 2.0 2 4.439 9.4 0.300

 4 3.7 2 4.439 9.4 1.100

 5 7.4 2 4.439 9.4 0.100

 6 15 2 4.439 9.4 1.300

------------------------------------------------------------------------------

F0 female length

File: 5401fl Transform: NO TRANSFORMATION

 WILLIAMS TEST (Isotonic regression model) TABLE 1 OF 2

----------------------------------------------------------------------------

GROUP ORIGINAL TRANSFORMED ISOTONIZED 

 IDENTIFICATION N MEAN MEAN MEAN 

------ -------------------- --- ----------- ----------- -----------

 1 control 2 47.400 47.400 47.400

 2 1.2 2 47.000 47.000 47.050

 3 2.0 2 47.100 47.100 47.050
DP Barcode: D306508 MRID No: 46315401

43

 4 3.7 2 46.300 46.300 46.800

 5 7.4 2 47.300 47.300 46.800

 6 15 2 46.100 46.100 46.100

----------------------------------------------------------------------------

F0 female length

File: 5401fl Transform: NO TRANSFORMATION

 WILLIAMS TEST (Isotonic regression model) TABLE 2 OF 2

----------------------------------------------------------------------------

 ISOTONIZED CALC. SIG TABLE DEGREES OF

 IDENTIFICATION MEAN WILLIAMS P=.05 WILLIAMS FREEDOM

-------------------- ----------- ----------- ----- ----------- -------------

 control 47.400

 1.2 47.050 0.223 1.94 k= 1, v= 6

 2.0 47.050 0.223 2.06 k= 2, v= 6

 3.7 46.800 0.383 2.10 k= 3, v= 6

 7.4 46.800 0.383 2.12 k= 4, v= 6

 15 46.100 0.829 2.13 k= 5, v= 6

----------------------------------------------------------------------------

s = 1.568

Note: df used for table values are approximate when v > 20.

F0 male weight

File: 5401mw Transform: NO TRANSFORMATION

 ANOVA TABLE

------------------------------------------------------------------------------

SOURCE DF SS MS F

------------------------------------------------------------------------------

Between 5 0.467 0.093 2.906

Within (Error) 6 0.190 0.032

------------------------------------------------------------------------------

Total 11 0.657
DP Barcode: D306508 MRID No: 46315401

44

------------------------------------------------------------------------------

 Critical F value = 4.39 (0.05,5,6)

 Since F < Critical F FAIL TO REJECT Ho:All groups equal

F0 male weight

File: 5401mw Transform: NO TRANSFORMATION

 DUNNETTS TEST - TABLE 1 OF 2 Ho:Control<Treatment

----------------------------------------------------------------------------

 TRANSFORMED MEAN CALCULATED IN 

GROUP IDENTIFICATION MEAN ORIGINAL UNITS T STAT SIG

----- -------------------- ----------- ------------------ ------ ---

 1 control 3.050 3.050

 2 1.2 3.050 3.050 0.000

3 2.0 3.050 3.050 0.000

 4 3.7 3.050 3.050 0.000

 5 7.4 3.150 3.150 -0.559

 6 15 2.550 2.550 2.795

----------------------------------------------------------------------------

Dunnett table value = 2.83 (1 Tailed Value, P=0.05, df=6,5)

F0 male weight

File: 5401mw Transform: NO TRANSFORMATION

 DUNNETTS TEST - TABLE 2 OF 2 Ho:Control<Treatment

----------------------------------------------------------------------------

 NUM OF Minimum Sig Diff % of DIFFERENCE

GROUP IDENTIFICATION REPS (IN ORIG. UNITS) CONTROL FROM CONTROL

----- -------------------- ------- ---------------- ------- ------------

 1 control 2

 2 1.2 2 0.506 16.6 0.000

 3 2.0 2 0.506 16.6 0.000

 4 3.7 2 0.506 16.6 0.000
DP Barcode: D306508 MRID No: 46315401

45

 5 7.4 2 0.506 16.6 -0.100

 6 15 2 0.506 16.6 0.500

------------------------------------------------------------------------------

F0 male weight

File: 5401mw Transform: NO TRANSFORMATION

 WILLIAMS TEST (Isotonic regression model) TABLE 1 OF 2

----------------------------------------------------------------------------

GROUP ORIGINAL TRANSFORMED ISOTONIZED 

 IDENTIFICATION N MEAN MEAN MEAN 

------ -------------------- --- ----------- ----------- -----------

 1 control 2 3.050 3.050 3.070

 2 1.2 2 3.050 3.050 3.070

 3 2.0 2 3.050 3.050 3.070

 4 3.7 2 3.050 3.050 3.070

 5 7.4 2 3.150 3.150 3.070

 6 15 2 2.550 2.550 2.550

----------------------------------------------------------------------------

F0 male weight

File: 5401mw Transform: NO TRANSFORMATION

 WILLIAMS TEST (Isotonic regression model) TABLE 2 OF 2

----------------------------------------------------------------------------

 ISOTONIZED CALC. SIG TABLE DEGREES OF

 IDENTIFICATION MEAN WILLIAMS P=.05 WILLIAMS FREEDOM

-------------------- ----------- ----------- ----- ----------- -------------

 control 3.070

 1.2 3.070 0.112 1.94 k= 1, v= 6

 2.0 3.070 0.112 2.06 k= 2, v= 6

 3.7 3.070 0.112 2.10 k= 3, v= 6

 7.4 3.070 0.112 2.12 k= 4, v= 6

 15 2.550 2.810 * 2.13 k= 5, v= 6
DP Barcode: D306508 MRID No: 46315401

46

----------------------------------------------------------------------------

s = 0.178

Note: df used for table values are approximate when v > 20.

F0 female weight

File: 5401fw Transform: NO TRANSFORM

 KRUSKAL-WALLIS ANOVA BY RANKS - TABLE 1 OF 2

----------------------------------------------------------------------------

 TRANSFORMED MEAN CALCULATED IN RANK 

GROUP IDENTIFICATION MEAN ORIGINAL UNITS SUM 

----- -------------------- ----------- ------------------ -----------

 1 control 2.100 2.100 17.500

 2 1.2 2.000 2.000 13.000

 3 2.0 2.050 2.050 14.500

 4 3.7 2.000 2.000 13.000

 5 7.4 2.000 2.000 14.000

 6 15 1.850 1.850 6.000

----------------------------------------------------------------------------

 Calculated H Value = 2.879 Critical H Value Table = 11.070

 Since Calc H < Crit H FAIL TO REJECT Ho:All groups are equal.

F0 female weight

File: 5401fw Transform: NO TRANSFORM

DUNNS MULTIPLE COMPARISON - KRUSKAL-WALLIS - TABLE 2 OF 2

----------------------------------------------------------------------------

 GROUP

 TRANSFORMED ORIGINAL 0 0 0 0 0 0 

GROUP IDENTIFICATION MEAN MEAN 6 4 5 2 3 1 

----- --------------- ----------- --------- - - - - - - 

 6 15 1.850 1.850 \

 4 3.7 2.000 2.000 . \

 5 7.4 2.000 2.000 . . \

 2 1.2 2.000 2.000 . . . \
DP Barcode: D306508 MRID No: 46315401

47

 3 2.0 2.050 2.050 . . . . \

 1 control 2.100 2.100 . . . . . \

----------------------------------------------------------------------------

* = significant difference (p=0.05) . = no significant difference

Table q value (0.05,6) = 2.936 SE = 3.548

total eggs

File: 5401e Transform: NO TRANSFORMATION

 ANOVA TABLE

------------------------------------------------------------------------------

SOURCE DF SS MS F

------------------------------------------------------------------------------

Between 5 1807373.750 361474.750 0.874

Within (Error) 6 2482450.500 413741.750

------------------------------------------------------------------------------

Total 11 4289824.250

------------------------------------------------------------------------------

 Critical F value = 4.39 (0.05,5,6)

 Since F < Critical F FAIL TO REJECT Ho:All groups equal

total eggs

File: 5401e Transform: NO TRANSFORMATION

 DUNNETTS TEST - TABLE 1 OF 2 Ho:Control<Treatment

----------------------------------------------------------------------------

 TRANSFORMED MEAN CALCULATED IN 

GROUP IDENTIFICATION MEAN ORIGINAL UNITS T STAT SIG

----- -------------------- ----------- ------------------ ------ ---

 1 control 3444.000 3444.000

 2 1.2 3311.000 3311.000 0.207

 3 2.0 3385.000 3385.000 0.092
DP Barcode: D306508 MRID No: 46315401

48

 4 3.7 4154.500 4154.500 -1.105

 5 7.4 3438.000 3438.000 0.009

 6 15 2828.000 2828.000 0.958

----------------------------------------------------------------------------

Dunnett table value = 2.83 (1 Tailed Value, P=0.05, df=6,5)

total eggs

File: 5401e Transform: NO TRANSFORMATION

 DUNNETTS TEST - TABLE 2 OF 2 Ho:Control<Treatment

----------------------------------------------------------------------------

 NUM OF Minimum Sig Diff % of DIFFERENCE

GROUP IDENTIFICATION REPS (IN ORIG. UNITS) CONTROL FROM CONTROL

----- -------------------- ------- ---------------- ------- ------------

 1 control 2

 2 1.2 2 1820.334 52.9 133.000

 3 2.0 2 1820.334 52.9 59.000

 4 3.7 2 1820.334 52.9 -710.500

 5 7.4 2 1820.334 52.9 6.000

 6 15 2 1820.334 52.9 616.000

------------------------------------------------------------------------------

total eggs

File: 5401e Transform: NO TRANSFORMATION

 WILLIAMS TEST (Isotonic regression model) TABLE 1 OF 2

----------------------------------------------------------------------------

GROUP ORIGINAL TRANSFORMED ISOTONIZED 

 IDENTIFICATION N MEAN MEAN MEAN 

------ -------------------- --- ----------- ----------- -----------

 1 control 2 3444.000 3444.000 3573.625

 2 1.2 2 3311.000 3311.000 3573.625

 3 2.0 2 3385.000 3385.000 3573.625

 4 3.7 2 4154.500 4154.500 3573.625
DP Barcode: D306508 MRID No: 46315401

49

 5 7.4 2 3438.000 3438.000 3438.000

 6 15 2 2828.000 2828.000 2828.000

----------------------------------------------------------------------------

total eggs

File: 5401e Transform: NO TRANSFORMATION

 WILLIAMS TEST (Isotonic regression model) TABLE 2 OF 2

----------------------------------------------------------------------------

 ISOTONIZED CALC. SIG TABLE DEGREES OF

 IDENTIFICATION MEAN WILLIAMS P=.05 WILLIAMS FREEDOM

-------------------- ----------- ----------- ----- ----------- -------------

 control 3573.625

 1.2 3573.625 0.202 1.94 k= 1, v= 6

 2.0 3573.625 0.202 2.06 k= 2, v= 6

 3.7 3573.625 0.202 2.10 k= 3, v= 6

 7.4 3438.000 0.009 2.12 k= 4, v= 6

 15 2828.000 0.958 2.13 k= 5, v= 6

----------------------------------------------------------------------------

s = 643.228

Note: df used for table values are approximate when v > 20.

eggs/female/day

File: 5401ed Transform: NO TRANSFORMATION

 ANOVA TABLE

------------------------------------------------------------------------------

SOURCE DF SS MS F

------------------------------------------------------------------------------

Between 5 74.750 14.950 0.790

Within (Error) 6 113.500 18.917

------------------------------------------------------------------------------

Total 11 188.250

------------------------------------------------------------------------------
DP Barcode: D306508 MRID No: 46315401

50

 Critical F value = 4.39 (0.05,5,6)

 Since F < Critical F FAIL TO REJECT Ho:All groups equal

eggs/female/day

File: 5401ed Transform: NO TRANSFORMATION

 DUNNETTS TEST - TABLE 1 OF 2 Ho:Control<Treatment

----------------------------------------------------------------------------

 TRANSFORMED MEAN CALCULATED IN 

GROUP IDENTIFICATION MEAN ORIGINAL UNITS T STAT SIG

----- -------------------- ----------- ------------------ ------ ---

 1 control 25.500 25.500

 2 1.2 24.500 24.500 0.230

 3 2.0 25.000 25.000 0.115

 4 3.7 30.000 30.000 -1.035

 5 7.4 25.000 25.000 0.115

 6 15 21.500 21.500 0.920

----------------------------------------------------------------------------

Dunnett table value = 2.83 (1 Tailed Value, P=0.05, df=6,5)

eggs/female/day

File: 5401ed Transform: NO TRANSFORMATION

 DUNNETTS TEST - TABLE 2 OF 2 Ho:Control<Treatment

----------------------------------------------------------------------------

 NUM OF Minimum Sig Diff % of DIFFERENCE

GROUP IDENTIFICATION REPS (IN ORIG. UNITS) CONTROL FROM CONTROL

----- -------------------- ------- ---------------- ------- ------------

 1 control 2

 2 1.2 2 12.309 48.3 1.000

 3 2.0 2 12.309 48.3 0.500

 4 3.7 2 12.309 48.3 -4.500

 5 7.4 2 12.309 48.3 0.500
DP Barcode: D306508 MRID No: 46315401

51

 6 15 2 12.309 48.3 4.000

------------------------------------------------------------------------------

eggs/female/day

File: 5401ed Transform: NO TRANSFORMATION

 WILLIAMS TEST (Isotonic regression model) TABLE 1 OF 2

----------------------------------------------------------------------------

GROUP ORIGINAL TRANSFORMED ISOTONIZED 

 IDENTIFICATION N MEAN MEAN MEAN 

------ -------------------- --- ----------- ----------- -----------

 1 control 2 25.500 25.500 26.250

 2 1.2 2 24.500 24.500 26.250

 3 2.0 2 25.000 25.000 26.250

 4 3.7 2 30.000 30.000 26.250

 5 7.4 2 25.000 25.000 25.000

 6 15 2 21.500 21.500 21.500

----------------------------------------------------------------------------

eggs/female/day

File: 5401ed Transform: NO TRANSFORMATION

 WILLIAMS TEST (Isotonic regression model) TABLE 2 OF 2

----------------------------------------------------------------------------

 ISOTONIZED CALC. SIG TABLE DEGREES OF

 IDENTIFICATION MEAN WILLIAMS P=.05 WILLIAMS FREEDOM

-------------------- ----------- ----------- ----- ----------- -------------

 control 26.250

 1.2 26.250 0.172 1.94 k= 1, v= 6

 2.0 26.250 0.172 2.06 k= 2, v= 6

 3.7 26.250 0.172 2.10 k= 3, v= 6

 7.4 25.000 0.115 2.12 k= 4, v= 6

 15 21.500 0.920 2.13 k= 5, v= 6

----------------------------------------------------------------------------
DP Barcode: D306508 MRID No: 46315401

52

s = 4.349

Note: df used for table values are approximate when v > 20.

hatching success F1

File: 5401h1 Transform: NO TRANSFORMATION

 ANOVA TABLE

------------------------------------------------------------------------------

SOURCE DF SS MS F

------------------------------------------------------------------------------

Between 5 616.417 123.283 1.702

Within (Error) 6 434.500 72.417

------------------------------------------------------------------------------

Total 11 1050.917

------------------------------------------------------------------------------

 Critical F value = 4.39 (0.05,5,6)

 Since F < Critical F FAIL TO REJECT Ho:All groups equal

hatching success F1

File: 5401h1 Transform: NO TRANSFORMATION

 DUNNETTS TEST - TABLE 1 OF 2 Ho:Control<Treatment

----------------------------------------------------------------------------

 TRANSFORMED MEAN CALCULATED IN 

GROUP IDENTIFICATION MEAN ORIGINAL UNITS T STAT SIG

----- -------------------- ----------- ------------------ ------ ---

 1 control 68.000 68.000

 2 1.2 59.000 59.000 1.058

 3 2.0 70.000 70.000 -0.235

 4 3.7 74.000 74.000 -0.705

 5 7.4 66.000 66.000 0.235

 6 15 52.500 52.500 1.821

----------------------------------------------------------------------------
DP Barcode: D306508 MRID No: 46315401

53

Dunnett table value = 2.83 (1 Tailed Value, P=0.05, df=6,5)

hatching success F1

File: 5401h1 Transform: NO TRANSFORMATION

 DUNNETTS TEST - TABLE 2 OF 2 Ho:Control<Treatment

----------------------------------------------------------------------------

 NUM OF Minimum Sig Diff % of DIFFERENCE

GROUP IDENTIFICATION REPS (IN ORIG. UNITS) CONTROL FROM CONTROL

----- -------------------- ------- ---------------- ------- ------------

 1 control 2

 2 1.2 2 24.083 35.4 9.000

 3 2.0 2 24.083 35.4 -2.000

 4 3.7 2 24.083 35.4 -6.000

 5 7.4 2 24.083 35.4 2.000

 6 15 2 24.083 35.4 15.500

------------------------------------------------------------------------------

hatching success F1

File: 5401h1 Transform: NO TRANSFORMATION

 WILLIAMS TEST (Isotonic regression model) TABLE 1 OF 2

----------------------------------------------------------------------------

GROUP ORIGINAL TRANSFORMED ISOTONIZED 

 IDENTIFICATION N MEAN MEAN MEAN 

------ -------------------- --- ----------- ----------- -----------

 1 control 2 68.000 68.000 68.000

 2 1.2 2 59.000 59.000 67.667

 3 2.0 2 70.000 70.000 67.667

 4 3.7 2 74.000 74.000 67.667

 5 7.4 2 66.000 66.000 66.000

 6 15 2 52.500 52.500 52.500

----------------------------------------------------------------------------
DP Barcode: D306508 MRID No: 46315401

54

hatching success F1

File: 5401h1 Transform: NO TRANSFORMATION

 WILLIAMS TEST (Isotonic regression model) TABLE 2 OF 2

----------------------------------------------------------------------------

 ISOTONIZED CALC. SIG TABLE DEGREES OF

 IDENTIFICATION MEAN WILLIAMS P=.05 WILLIAMS FREEDOM

-------------------- ----------- ----------- ----- ----------- -------------

 control 68.000

 1.2 67.667 0.039 1.94 k= 1, v= 6

 2.0 67.667 0.039 2.06 k= 2, v= 6

 3.7 67.667 0.039 2.10 k= 3, v= 6

 7.4 66.000 0.235 2.12 k= 4, v= 6

 15 52.500 1.821 2.13 k= 5, v= 6

----------------------------------------------------------------------------

s = 8.510

Note: df used for table values are approximate when v > 20.

survival F1

File: 5401s1 Transform: NO TRANSFORM

 KRUSKAL-WALLIS ANOVA BY RANKS - TABLE 1 OF 2

----------------------------------------------------------------------------

 TRANSFORMED MEAN CALCULATED IN RANK 

GROUP IDENTIFICATION MEAN ORIGINAL UNITS SUM 

----- -------------------- ----------- ------------------ -----------

 1 control 95.000 95.000 12.000

 2 1.2 95.000 95.000 10.500

 3 2.0 98.000 98.000 15.500

 4 3.7 93.000 93.000 6.500

 5 7.4 100.000 100.000 21.000

 6 15 97.000 97.000 12.500

----------------------------------------------------------------------------

 Calculated H Value = 4.889 Critical H Value Table = 11.070

 Since Calc H < Crit H FAIL TO REJECT Ho:All groups are equal.
DP Barcode: D306508 MRID No: 46315401

55

survival F1

File: 5401s1 Transform: NO TRANSFORM

DUNNS MULTIPLE COMPARISON - KRUSKAL-WALLIS - TABLE 2 OF 2

----------------------------------------------------------------------------

 GROUP

 TRANSFORMED ORIGINAL 0 0 0 0 0 0 

GROUP IDENTIFICATION MEAN MEAN 4 1 2 6 3 5 

----- --------------- ----------- --------- - - - - - - 

 4 3.7 93.000 93.000 \

 1 control 95.000 95.000 . \

 2 1.2 95.000 95.000 . . \

 6 15 97.000 97.000 . . . \

 3 2.0 98.000 98.000 . . . . \

 5 7.4 100.000 100.000 . . . . . \

----------------------------------------------------------------------------

* = significant difference (p=0.05) . = no significant difference

Table q value (0.05,6) = 2.936 SE = 3.503

length F1

File: 5401l1 Transform: NO TRANSFORMATION

 ANOVA TABLE

------------------------------------------------------------------------------

SOURCE DF SS MS F

------------------------------------------------------------------------------

Between 5 27.064 5.413 0.935

Within (Error) 6 34.725 5.787

------------------------------------------------------------------------------

Total 11 61.789

------------------------------------------------------------------------------
DP Barcode: D306508 MRID No: 46315401

56

 Critical F value = 4.39 (0.05,5,6)

 Since F < Critical F FAIL TO REJECT Ho:All groups equal

length F1

File: 5401l1 Transform: NO TRANSFORMATION

 DUNNETTS TEST - TABLE 1 OF 2 Ho:Control<Treatment

----------------------------------------------------------------------------

 TRANSFORMED MEAN CALCULATED IN 

GROUP IDENTIFICATION MEAN ORIGINAL UNITS T STAT SIG

----- -------------------- ----------- ------------------ ------ ---

 1 control 25.050 25.050

 2 1.2 21.000 21.000 1.684

 3 2.0 24.950 24.950 0.042

 4 3.7 22.650 22.650 0.998

 5 7.4 22.450 22.450 1.081

 6 15 21.950 21.950 1.289

----------------------------------------------------------------------------

Dunnett table value = 2.83 (1 Tailed Value, P=0.05, df=6,5)

length F1

File: 5401l1 Transform: NO TRANSFORMATION

 DUNNETTS TEST - TABLE 2 OF 2 Ho:Control<Treatment

----------------------------------------------------------------------------

 NUM OF Minimum Sig Diff % of DIFFERENCE

GROUP IDENTIFICATION REPS (IN ORIG. UNITS) CONTROL FROM CONTROL

----- -------------------- ------- ---------------- ------- ------------

 1 control 2

 2 1.2 2 6.808 27.2 4.050

 3 2.0 2 6.808 27.2 0.100

 4 3.7 2 6.808 27.2 2.400

 5 7.4 2 6.808 27.2 2.600

 6 15 2 6.808 27.2 3.100
DP Barcode: D306508 MRID No: 46315401

57

------------------------------------------------------------------------------

length F1

File: 5401l1 Transform: NO TRANSFORMATION

 WILLIAMS TEST (Isotonic regression model) TABLE 1 OF 2

----------------------------------------------------------------------------

GROUP ORIGINAL TRANSFORMED ISOTONIZED 

 IDENTIFICATION N MEAN MEAN MEAN 

------ -------------------- --- ----------- ----------- -----------

 1 control 2 25.050 25.050 25.050

 2 1.2 2 21.000 21.000 22.975

 3 2.0 2 24.950 24.950 22.975

 4 3.7 2 22.650 22.650 22.650

 5 7.4 2 22.450 22.450 22.450

 6 15 2 21.950 21.950 21.950

----------------------------------------------------------------------------

length F1

File: 5401l1 Transform: NO TRANSFORMATION

 WILLIAMS TEST (Isotonic regression model) TABLE 2 OF 2

----------------------------------------------------------------------------

 ISOTONIZED CALC. SIG TABLE DEGREES OF

 IDENTIFICATION MEAN WILLIAMS P=.05 WILLIAMS FREEDOM

-------------------- ----------- ----------- ----- ----------- -------------

 control 25.050

 1.2 22.975 0.863 1.94 k= 1, v= 6

 2.0 22.975 0.863 2.06 k= 2, v= 6

 3.7 22.650 0.998 2.10 k= 3, v= 6

 7.4 22.450 1.081 2.12 k= 4, v= 6

 15 21.950 1.289 2.13 k= 5, v= 6

----------------------------------------------------------------------------

s = 2.406
DP Barcode: D306508 MRID No: 46315401

58

Note: df used for table values are approximate when v > 20.

weight F1

File: 5401w1 Transform: NO TRANSFORMATION

 KRUSKAL-WALLIS ANOVA BY RANKS - TABLE 1 OF 2

----------------------------------------------------------------------------

 TRANSFORMED MEAN CALCULATED IN RANK 

GROUP IDENTIFICATION MEAN ORIGINAL UNITS SUM 

----- -------------------- ----------- ------------------ -----------

 1 control 256.000 256.000 21.000

 2 1.2 157.150 157.150 9.500

 3 2.0 251.000 251.000 20.500

 4 3.7 175.000 175.000 8.500

 5 7.4 175.500 175.500 9.500

 6 15 173.000 173.000 9.000

----------------------------------------------------------------------------

 Calculated H Value = 7.035 Critical H Value Table = 11.070

 Since Calc H < Crit H FAIL TO REJECT Ho:All groups are equal.

weight F1

File: 5401w1 Transform: NO TRANSFORMATION

DUNNS MULTIPLE COMPARISON - KRUSKAL-WALLIS - TABLE 2 OF 2

----------------------------------------------------------------------------

 GROUP

 TRANSFORMED ORIGINAL 0 0 0 0 0 0 

GROUP IDENTIFICATION MEAN MEAN 2 6 4 5 3 1 

----- --------------- ----------- --------- - - - - - - 

 2 1.2 157.150 157.150 \

 6 15 173.000 173.000 . \

 4 3.7 175.000 175.000 . . \

 5 7.4 175.500 175.500 . . . \

 3 2.0 251.000 251.000 . . . . \

 1 control 256.000 256.000 . . . . . \
DP Barcode: D306508 MRID No: 46315401

59

----------------------------------------------------------------------------

* = significant difference (p=0.05) . = no significant difference

Table q value (0.05,6) = 2.936 SE = 3.587

%hatch 6 days

File: 5401h6 Transform: NO TRANSFORMATION

 KRUSKAL-WALLIS ANOVA BY RANKS - TABLE 1 OF 2

----------------------------------------------------------------------------

 TRANSFORMED MEAN CALCULATED IN RANK 

GROUP IDENTIFICATION MEAN ORIGINAL UNITS SUM 

----- -------------------- ----------- ------------------ -----------

 1 control 0.000 0.000 9.000

 2 1.2 5.500 5.500 19.000

 3 2.0 25.000 25.000 16.500

 4 3.7 0.000 0.000 9.000

 5 7.4 0.000 0.000 9.000

 6 15 7.500 7.500 15.500

----------------------------------------------------------------------------

 Calculated H Value = 5.582 Critical H Value Table = 11.070

 Since Calc H < Crit H FAIL TO REJECT Ho:All groups are equal.

%hatch 6 days

File: 5401h6 Transform: NO TRANSFORMATION

DUNNS MULTIPLE COMPARISON - KRUSKAL-WALLIS - TABLE 2 OF 2

----------------------------------------------------------------------------

 GROUP

 TRANSFORMED ORIGINAL 0 0 0 0 0 0 

GROUP IDENTIFICATION MEAN MEAN 1 4 5 2 6 3 

----- --------------- ----------- --------- - - - - - - 

 1 control 0.000 0.000 \

 4 3.7 0.000 0.000 . \

 5 7.4 0.000 0.000 . . \
DP Barcode: D306508 MRID No: 46315401

60

 2 1.2 5.500 5.500 . . . \

 6 15 7.500 7.500 . . . . \

 3 2.0 25.000 25.000 . . . . . \

----------------------------------------------------------------------------

* = significant difference (p=0.05) . = no significant difference

Table q value (0.05,6) = 2.936 SE = 3.030

#hatch 7 days

File: 5401h7 Transform: NO TRANSFORMATION

 ANOVA TABLE

------------------------------------------------------------------------------

SOURCE DF SS MS F

------------------------------------------------------------------------------

Between 5 889.417 177.883 0.407

Within (Error) 6 2619.500 436.583

------------------------------------------------------------------------------

Total 11 3508.917

------------------------------------------------------------------------------

 Critical F value = 4.39 (0.05,5,6)

 Since F < Critical F FAIL TO REJECT Ho:All groups equal

#hatch 7 days

File: 5401h7 Transform: NO TRANSFORMATION

 DUNNETTS TEST - TABLE 1 OF 2 Ho:Control<Treatment

----------------------------------------------------------------------------

 TRANSFORMED MEAN CALCULATED IN 

GROUP IDENTIFICATION MEAN ORIGINAL UNITS T STAT SIG

----- -------------------- ----------- ------------------ ------ ---

 1 control 94.500 94.500

 2 1.2 92.000 92.000 0.120
DP Barcode: D306508 MRID No: 46315401

61

 3 2.0 71.000 71.000 1.125

 4 3.7 94.000 94.000 0.024

 5 7.4 81.500 81.500 0.622

 6 15 92.500 92.500 0.096

----------------------------------------------------------------------------

Dunnett table value = 2.83 (1 Tailed Value, P=0.05, df=6,5)

#hatch 7 days

File: 5401h7 Transform: NO TRANSFORMATION

 DUNNETTS TEST - TABLE 2 OF 2 Ho:Control<Treatment

----------------------------------------------------------------------------

 NUM OF Minimum Sig Diff % of DIFFERENCE

GROUP IDENTIFICATION REPS (IN ORIG. UNITS) CONTROL FROM CONTROL

----- -------------------- ------- ---------------- ------- ------------

 1 control 2

 2 1.2 2 59.132 62.6 2.500

 3 2.0 2 59.132 62.6 23.500

 4 3.7 2 59.132 62.6 0.500

 5 7.4 2 59.132 62.6 13.000

 6 15 2 59.132 62.6 2.000

------------------------------------------------------------------------------

#hatch 7 days

File: 5401h7 Transform: NO TRANSFORMATION

 WILLIAMS TEST (Isotonic regression model) TABLE 1 OF 2

----------------------------------------------------------------------------

GROUP ORIGINAL TRANSFORMED ISOTONIZED 

 IDENTIFICATION N MEAN MEAN MEAN 

------ -------------------- --- ----------- ----------- -----------

 1 control 2 94.500 94.500 94.500

 2 1.2 2 92.000 92.000 92.000

 3 2.0 2 71.000 71.000 84.750
DP Barcode: D306508 MRID No: 46315401

62

 4 3.7 2 94.000 94.000 84.750

 5 7.4 2 81.500 81.500 84.750

 6 15 2 92.500 92.500 84.750

----------------------------------------------------------------------------

#hatch 7 days

File: 5401h7 Transform: NO TRANSFORMATION

 WILLIAMS TEST (Isotonic regression model) TABLE 2 OF 2

----------------------------------------------------------------------------

 ISOTONIZED CALC. SIG TABLE DEGREES OF

 IDENTIFICATION MEAN WILLIAMS P=.05 WILLIAMS FREEDOM

-------------------- ----------- ----------- ----- ----------- -------------

 control 94.500

 1.2 92.000 0.120 1.94 k= 1, v= 6

 2.0 84.750 0.467 2.06 k= 2, v= 6

 3.7 84.750 0.467 2.10 k= 3, v= 6

 7.4 84.750 0.467 2.12 k= 4, v= 6

 15 84.750 0.467 2.13 k= 5, v= 6

----------------------------------------------------------------------------

s = 20.895

Note: df used for table values are approximate when v > 20.

%hatch 8 days

File: 5401h8 Transform: NO TRANSFORMATION

 KRUSKAL-WALLIS ANOVA BY RANKS - TABLE 1 OF 2

----------------------------------------------------------------------------

 TRANSFORMED MEAN CALCULATED IN RANK 

GROUP IDENTIFICATION MEAN ORIGINAL UNITS SUM 

----- -------------------- ----------- ------------------ -----------

 1 control 5.500 5.500 14.500

 2 1.2 2.500 2.500 13.500

 3 2.0 0.000 0.000 9.000

 4 3.7 6.500 6.500 15.500

 5 7.4 19.000 19.000 16.500
DP Barcode: D306508 MRID No: 46315401

63

 6 15 0.000 0.000 9.000

----------------------------------------------------------------------------

 Calculated H Value = 2.886 Critical H Value Table = 11.070

 Since Calc H < Crit H FAIL TO REJECT Ho:All groups are equal.

%hatch 8 days

File: 5401h8 Transform: NO TRANSFORMATION

DUNNS MULTIPLE COMPARISON - KRUSKAL-WALLIS - TABLE 2 OF 2

----------------------------------------------------------------------------

 GROUP

 TRANSFORMED ORIGINAL 0 0 0 0 0 0 

GROUP IDENTIFICATION MEAN MEAN 3 6 2 1 4 5 

----- --------------- ----------- --------- - - - - - - 

 3 2.0 0.000 0.000 \

 6 15 0.000 0.000 . \

 2 1.2 2.500 2.500 . . \

 1 control 5.500 5.500 . . . \

 4 3.7 6.500 6.500 . . . . \

 5 7.4 19.000 19.000 . . . . . \

----------------------------------------------------------------------------

* = significant difference (p=0.05) . = no significant difference

Table q value (0.05,6) = 2.936 SE = 3.030