Registration Dossier

Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
3 February 1998 - 7 February 1998
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study was conducted in compliance with OECD and EPA guidelines. Restriction due to no analytical monitoring of test material.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1998
Report Date:
1998

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OTS 797.1400 (Fish Acute Toxicity Test)
Deviations:
no
Principles of method if other than guideline:
Not applicable.
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
Not applicable.

Sampling and analysis

Analytical monitoring:
no
Details on sampling:
- Concentrations: 0, 0.11, 0.35, 3.35, 33.02, 330 mg/L

- Sampling method: TOC measured in each test treatment after 24 hour exposures. Total Organic Carbon per APHA 1992 Standard Methods for the Examination of Water and Wastewater, 18th Edition

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)

- Method: Oil-Water dispersion prepared by adding a measured volume of product to a known volume of water. The product was added to the test chamber within the confines of a glass mixing chamber. The chamber was constructed with holes from the bottom of the chamber and below the surface of the water. Holes were covered with Nytex screen to prevent fish from entering the mixing chamber. A stirring motor with polyethylene covered shaft and mixing blade were centered over the mixing chamber. The product was allowed to mix for 1 hour prior to adding the fish. Mixing was done at approximately 1000 rpm.

- Controls: untreated negative control

- Chemical name of vehicle (organic solvent, emulsifier or dispersant): reconstituted water

- Evidence of undissolved material (e.g. precipitate, surface film, etc): Concentrations of 0.11 and 0.35 mg/L had no noticeable undissolved test material present. Some product was observed at the bottom of the tank and/or as a film on the water surface in the 3.35 mg/L, 33.02, and 330 mg/L concentrations.

Test organisms

Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
TEST ORGANISM

- Common name: Rainbow trout

- Strain: Oncorhynchus mykiss identified using appropriate taxonomic key (Eddy 1969)

- Source: Single source from in house cultures

- Age at study initiation (mean and range, SD): juveniles

- Length at study initiation (length definition, mean, range and SD): range 21.5 to 33.1 mm; mean 27.5 mm

- Weight at study initiation (mean and range, SD): range 0.1038 to 0.4728 grams; mean weight of 0.2453 grams

- Method of breeding: Fish were maintained in a static renewal or flow through condition culture system prior to test initiation as appropriate and in the same water and at the same photoperiod and temperature that was used for the assay. Pretest observation data was recorded concerning the source, handling procedures, receipt date, disease treatment (if any), health, feeding and mortality.

FEEDING DURING TEST

- Food type: commercial flake food

- Amount: no data available

- Frequency: once per day up to the start of the test


ACCLIMATION

- Acclimation period: 14 days

- Acclimation conditions (same as test or not): Yes same as test. Fish were maintained in a static renewal or flow through condition culture system prior to test initiation as appropriate and in the same water and at the same photoperiod and temperature that was used for the assay. There was a 14 day acclimation period prior to starting the assay and the fish were fed during this period.

- Type and amount of food: no data available

- Feeding frequency: no data available

- Health during acclimation (any mortality observed): Pretest observation data was recorded concerning the source, handling procedures, receipt date, disease treatment (if any), health, feeding and mortality.


QUARANTINE (wild caught)

- Duration: not applicable

- Health/mortality: not applicable



Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h

Test conditions

Hardness:
171 mg/L as CaCO3 in reconstituted water controls, 176 mg/L as CaCO3 in test concentrations
Test temperature:
Mean temperature was 15.7 deg. C
pH:
pH values in the control and in the test concentrations ranged from 7.8 to 8.01. No treatment related differences in pH
Dissolved oxygen:
DO levels in controls and test concentrations ranged from 95% to 100% saturation. No treatment related differences in DO
Salinity:
Not applicable
Nominal and measured concentrations:
Nominal concentrations of 0, 0.11, 0.35, 3.35, 33.02, and 330 mg/L; concentration gradient verified by TOC analysis
Details on test conditions:
TEST SYSTEM
- Test vessel: 5 gallon aquaria, each containing 15L of test solution

- Material, size, headspace, fill volume: 15 L of test solution

- Aeration: not aerated

- Renewal rate of test solution (frequency/flow rate): Static

- No. of organisms per vessel: 10

- No. of vessels per concentration (replicates): 1

- No. of vessels per control (replicates): 1

- Biomass loading rate: Test animals were equally and randomly assigned to the test vessels; The loading rate was determined to be 0.16 g/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted water per EPA 1993 specifications

- Total organic carbon: 0.9 mg/L

- Particulate matter: Total Solids at 368 mg/L

- Metals: no data available

- Pesticides: no data available

- Chlorine: no data available

- Alkalinity: 110 mg/L CaCO3

- Ca/Mg ratio: Hardness: 171 mg/L CaCO3

- Conductivity: ranged from 467 to 505 umhos/cm in controls

- Culture medium different from test medium: No difference, the water in the culture system was the same type of water used in the bioassay. Fish were acclimated to the reconstituted water for a minimum of 14 days prior to testing.


OTHER TEST CONDITIONS

- Adjustment of pH: none

- Photoperiod: 16 hours light and 8 hours dark with 15-30 minute transition period

- Light intensity: ambient laboratory lighting, cool white fluorescent lights providing a light intensity of 60 foot candles


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality with notation of sublethal effects


TEST CONCENTRATIONS

- Spacing factor for test concentrations: roughly 10x

- Justification for using less concentrations than requested by guideline: NA

- Range finding study not conducted

- Test concentrations: 0.11, 0.35, 3.35, 33.02, 330 mg/L nominal concentrations

- Intervals of water quality measurement: Daily for temperature, pH, DO, and conductivity; TOC at 24 hours; Hardness, alkalinity, and total solids at 0 hours















Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
44.35 mg/L
Nominal / measured:
nominal
Conc. based on:
other: Nominal with concentration gradient confirmed by TOC
Basis for effect:
mortality
Remarks:
sublethal effects, if any, will be noted
Remarks on result:
other: 19.9-98.5 mg/L
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
33.02 mg/L
Nominal / measured:
nominal
Conc. based on:
other: nominal with concentration gradient confirmed by TOC
Basis for effect:
mortality
Details on results:
- Behavioural abnormalities: no sign of sublethal effects observed in controls. No sublethal effects were observed in test treatments during the 72 hour testing period. At the 96 hour exposure period, the sublethal effect of lethargy was observed in one organism in the 0.11 mg/L concentration.

- Other biological observations: no sign of sublethal effects

- Mortality of control: 100 percent survival

- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: Some product was observed on the water surface or on the bottom of the tank at the 3.35, 33.02 and 330 mg/L concentrations
Results with reference substance (positive control):
No reference tox reported
Reported statistics and error estimates:
Results of the toxicity test were interpreted using standard statistical techniques. LC50 values were computed using an EPA (Stephen 1983) computer program that uses the probit, spearman karber, and binomial methods.

Any other information on results incl. tables

Table 1: Survival and Sublethal Effect Data

Test Concentration mg/L

Replicate

Number of Survivors

at Hour

Number Affected

 at Hour

0

24

48

72

96

24

48

72

96

0.00

A

10

10

10

10

10

0

0

0

0

0.11

A

10

10

10

10

10

0

0

0

1

0.35

A

10

10

10

9

9

0

0

0

0

3.35

A

10

10

9

9

8

0

0

0

0

33.02

A

10

10

10

10

9

0

0

0

0

330

A

10

0

0

0

0

*

*

*

*

Table 2: Median Lethal Concentration (LC50), Median Effect Concentration (EC50), and No Observed Effect Concentration (NOEC)

Exposure Period (hours)

LC50 (mg/L)

EC50 (mg/L)

NOEC (mg/L)

24

104.69

104.69

33.02

48

83.07

83.07

33.02

72

68.69

68.69

33.02

96

44.35

44.35

33.02

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The acute toxicity of the test material to the freshwater fish, Oncorhynchus mykiss, was investigated and resulted in a 96 hour LC50 of 44.35 mg/L and an NOEC of 33.02 mg/L.
Executive summary:

Introduction. A study was performed to assess the acute toxicity of the test material to rainbow trout using OECD 203 and U.S. EPA 797 Guidelines.

 

Methods. 10 fish per test concentration were exposed to dispersions of the test material in reconstituted water over a range of nominal concentrations of 0, 0.11, 0.35, 3.35, 33.02, and 330 mg/L. TOC analysis was conducted that confirmed the concentration gradient. The test was conducted under static conditions for 96 hours at a temperature of 15C. The number of fish mortalities was recorded after, 24, 48, 72, and 96 hours. 

 

Results The acute toxicity of the test material to the freshwater fish, Oncorhynchus mykiss, was investigated and resulted in a 96 hour LC50 of 44.35 mg/L and an NOEC of 33.02 mg/L. No sublethal effects were observed during the 72 hour testing period in all treatments and controls. The sublethal effect of lethargy was observed in one organism in the 0.11 mg/L test concentration at the 96 hour exposure period.