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Short-term toxicity to fish

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Reference
Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From June 9 to 25, 2005
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
This study was performed according to OECD Guideline 203 and EU Method C.1 with GLP certificate. All validity criteria were fulfilled
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes (incl. certificate)
Remarks:
UK GLP Compliance Programme (inspected on April 12, 2005/ signed on June 01, 2005)
Analytical monitoring:
yes
Details on sampling:
- Concentrations: The aqueous samples were diluted with methanol and/or water to bring the expected analyte concentration within the calibration range of 0 – 1.0 mg/L.
- Sampling method: Samples of 100 mL of media were taken from the control and test vessels at 0, 24, 72 and 96 hours for analysis.
- Sample storage conditions before analysis: Sample were analyzed within two days of sampling and /or stored in a refrigerator in case further analysis was required.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances):
On each occasion, the test substance (nominally 20, 21, 47, 103, 227 or 500 mg) was added directly to volumetric flasks containing diluent water and made up to volume (20 to 227 mg into two litres or 500 mg into five litres). The contents of the flasks were shaken vigorously, treated by ultrasound for 30 minutes with periods of intermittent vigorous shaking, and stirred overnight in darkness. On cessation of stirring, an aliquot (970 mL) of the medium at nominaly 10 mg/L was added to a test vessel (15 litre all glass aquarium) and the volume adjusted to 10 litres with diluent water to provide the medium at 0.970 mg/L. The contents of each of the remaining flasks were added to a test vessel and the volume adjusted to 10 litres to provide the media at 2.13 to 50 mg/L.

Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
TEST ORGANISM
- Common name: Rainbow trout (Oncorhynchus mykiss)
- Source: Commercial trout farm in the UK
- Age at study initiation: no data available
- Length at study initiation (mean): 4.07 cm
- Weight at study initiation (mean): 0.89 g
- Method of breeding: no data available
- Feeding during test: No food was given during the 19-hour period immediately before exposure or during the exposure period itself.
- Food type: not applicable
- Amount: not applicable
- Frequency: not applicable

ACCLIMATION
- Acclimation period: 14 days
- Acclimation conditions: During the 14-day period immediately before the definitive test, temperatures remained within the range 14.1-15.4 °C, pH values within the range 7.55-8.05, dissolved oxygen concentrations within the range 87-100% air saturation value (ASV) and total hardness within the range 176-210 mg/L as CaCO3.
- Feeding: The fish were fed daily with commercial fish food (TROUW (UK) Ltd; Nutra Fry 02) an amount equivalent to between 1 and 3% of the total wet-weight of fish in the holding tank.
- Health during acclimation (any mortality observed): No medication was given during the holding period, and no mortality was recorded in the 14 days before the definitive test.
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Post exposure observation period:
None
Hardness:
176-210 mg/L as CaCo3
Test temperature:
15.9-17.1 °C
pH:
7.94-8.5
Dissolved oxygen:
within the range 89 to 100% air saturation value (ASV)
Salinity:
See Table 6.1.1/1: Typical water quality characteristics of the diluent supply
Nominal and measured concentrations:
Nominal concentrations: 0.97, 2.13, 4.70, 10.3, 22.7 and 50 mg/L
Arithmetic mean measured concentrations: 0.946, 1.97, 4.29, 9.49, 21.7 and 47.3 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: Glass aquarium containing the prepared test or control media. Each vessel contained 10 litres of medium to a depth of 15 cm. This provided an initial static loading of 0.62 g bodyweight/litre.
- Aeration: Supplementary aeration was provided via narrow bore glass tubes.
- Renewal rate of test solution (frequency/flow rate): The fish were exposed to the control or test conditions for a period of 96 hours with daily batch renewal of the media.
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per vehicle control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: The water used to hold the fish and for the study was laboratory tap water, dechlorinated and softened by passage through a water purification system (Vivendi Water Systems; formerly Elga).
- Water quality characteristics: See Table 6.1.1/1: Typical water quality characteristics of the diluent supply

OTHER TEST CONDITIONS
- Photoperiod: A photoperiod of 16 hours light: 8 hours dark was maintained, with periods of subdued lighting at the beginning and end of each light phase.
- Daily records of temperature, pH and dissolved oxygen were kept for each control and test vessel together with measurements of total hardness for selected vessels at 0 hours.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Observations on mortality at approximately 15 minutes, 2, 4, 24, 48, 72 and 96 hours and subjective assessments were also made on the incidence and type of any sub-lethal effects compared with control fish.

TEST CONCENTRATIONS
- Range finding study: The range finding test was conducted with test concentrations of 1, 10 and 100 mg/L.
- Results used to determine the conditions for the definitive study: Sub-lethal effects on the fish were observed in the 1 and 10 mg/L test groups however, the effects observed at 1 mg/L were not considered to be treatment related. As a result, the definitive test concentrations, which were selected based on the results of the range finding test, were: Nominal concentrations: 0.970, 2.13, 4.70, 10.3, 22.7 and 50 mg/L.
Reference substance (positive control):
no
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
14.4 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: 95% confidence limits of 9.49 and 21.7 mg/L
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
< 0.946 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: As significant sub-lethal effects were observed in all of the test groups, hence the NOEC was not identified.
Duration:
96 h
Dose descriptor:
LC0
Effect conc.:
9.49 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
21.7 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
After 96 hours, the highest measured concentration at which no mortality had occurred was 9.49 mg/L and the lowest at which there was 100% mortality was 21.7 mg/L. Treatment-related effects were exhibited at 0.946 mg/L and higher concentrations.
The following sub-lethal effects were recorded:
Conc. 0.946 mg/L: Darkened pigmentation (body and eye orbits), coughing
Conc. 1.97 mg/L: Nervous erratic behavior, Hyperventilation, Coughing, Quiescence
Conc. 4.29 mg/L: Hyperventilation, Coughing, Nervous erratic behaviotir, Darkened pigmentation (body and eye orbits), Quiescence
Conc. 9.49 mg/L: Hyperventilation, Nervous erratic behavior, Darkened pigmentation (body and eye orbits), Reduced opercular movement, Coughing, Quiescence, Loss of orientation (nose pointing downwards)
Conc. 21.7 mg/L: Overturned and immobile (base of vessel), Darkened pigmentation (body and eye orbits)
Results with reference substance (positive control):
Not applicable
Reported statistics and error estimates:
No additional data available

The measured concentrations of test item ranged between 84 and 106% of their nominal values in samples of freshly prepared media and expired (24 hours old) media during the test, giving overall arithmetic mean measured levels of 0.946, 1.97, 4.29, 9.49, 21.7 and 47.3 mg/L.

 

Table 6.1.1/2: Cumulative mortality

 

ST 06 C 05 (mg/L)

Cumulative mortality (initial population = 7 fish/concentration)

nominal

measured

0.25 h

2 h

4 h

24 h

48 h

72 h

96 h

%

Control

nd

0

0

0

0

0

0

0

0

0.970

0.946

0

0

0

0

0

0

0

0

2.13

1.97

0

0

0

0

0

0

0

0

4.70

4.29

0

0

0

0

0

0

0

0

10.3

9.49

0

0

0

0

0

0

0

0

22.7

21.7

0

4

7

7

7

7

7

100

50

47.3

7

7

7

7

7

7

7

100

 

nd: none detected (< 0.01 mg/L).

Validity criteria fulfilled:
yes
Conclusions:
The 96-hour LC50 for test item was 14.4 mg/L, with 95% confidence limits of 9.49 and 21.7 mg/L. No observed effect concentration was < 0.946 mg/L.
Executive summary:

The acute toxicity of the test item to Rainbow trout (Oncorhynchus mykiss) was determined in a 96-hour semi-static test according to EU Method C.1 and the OECD Guideline No. 203.

Groups of seven juvenile fish were exposed to test item, dissolved in water at nominal concentrations of 0.970, 2.13, 4.70, 10.3, 22.7 and 50 mg/L. The test media were prepared by direct addition of the test substance to water; to aid dissolution, ultrasound treatment and overnight stirring were employed. The measured concentrations of test item ranged between 84 and 106% of their nominal values in samples of freshly prepared media and expired (24 hours old) media during the test, giving overall arithmetic mean measured levels of 0.946, 1.97, 4.29, 9.49, 21.7 and 47.3 mg/L. Observations of the fish were made after approximately 0.25, 2, 4, 24, 48, 72 and 96 hours of exposure.

 

After 96 hours, the highest measured concentration at which no mortality had occurred was 9.49 mg/L and the lowest at which there was 100% mortality was 21.7 mg/L. Treatment-related effects were exhibited at 0.946 mg/L and higher concentrations.

 

Under the test conditions, the 96-hour LC50 for test item was 14.4 mg/L, with 95% confidence limits of 9.49 and 21.7 mg/L. No observed effect concentration was < 0.946 mg/L.

Description of key information

OECD Guideline 203, GLP, key study, validity 1:

96h-LC50 (Oncorhynchus mykiss) = 14.4 mg/L based on measured concentrations.

Key value for chemical safety assessment

LC50 for freshwater fish:
14.4 mg/L

Additional information

The acute toxicity of the test item toRainbow trout (Oncorhynchus mykiss)was determined in a 96-hour semi-static test according to EU Method C.1 and the OECD Guideline No. 203.

Groups of seven juvenile fish were exposed to test item, dissolved in water at nominal concentrations of 0.970, 2.13, 4.70, 10.3, 22.7 and 50 mg/L. The test media were prepared by direct addition of the test substance to water; to aid dissolution, ultrasound treatment and overnight stirring were employed. The measured concentrations of test item ranged between 84 and 106% of their nominal values in samples of freshly prepared media and expired (24 hours old) media during the test, giving overall arithmetic mean measured levels of 0.946, 1.97, 4.29, 9.49, 21.7 and 47.3 mg/L. Observations of the fish were made after approximately 0.25, 2, 4, 24, 48, 72 and 96 hours of exposure.

 

After 96 hours, the highest measured concentration at which no mortality had occurred was 9.49 mg/L and the lowest at which there was 100% mortality was 21.7 mg/L. Treatment-related effects were exhibited at 0.946 mg/L and higher concentrations.

 

Under the test conditions, the 96-hour LC50 for test item was 14.4 mg/L, with 95% confidence limits of 9.49 and 21.7 mg/L. No observed effect concentration was < 0.946 mg/L.