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Ecotoxicological information

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:
1983-06-03 to 1984-05-03
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Version / remarks:
version: adopted 12 May 1981
GLP compliance:
yes
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 153006
- Purity test date: 97.7%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material:
-- sample 1: in the dark at ambient temperature
-- sample 2: wrapped in aluminium at 4°C
Analytical monitoring:
yes
Details on sampling:
- Concentrations: control, 560, 1000, 1800 and 2400 mg/L
- Sampling method: samples were collected from each vessel at the beginning and the end of the test
- Sample storage conditions before analysis: frozen
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The appropriate weight of the test substance was placed into a volumetric flask and borehole water added (1 L). The content of the flask was equally divided and added to two vessels already containing 5 L borehole water. The preparation flasks were rinsed with water and the rinsing divided equally between the test vessels. Finally the volume of solution in each test vessel was made up to 10 L.
- Controls: 10 L dilution water was added directly to the control vessels
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): minute black particles were observed in concentrations 1800 mg/L and 2400 mg/L
Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
TEST ORGANISM
- Common name: Rainbow trout
- Source:
-- for preliminary and first definitive test: WRC Stevenage laboratory;
-- for second definitive test: Padworth Fisheries, Berkshire
- Weight at study initiation (mean of 10 fish): preliminary test: 0.85 g; first definitive test: 0.63 g; second definitive test: 0.86 g
- Maintenance of the brood fish: daily feeding with trout pellets (EWOS-BAKER Ltd., Size 2 or BP Nutrition Ltd, Mainstream Trout Fry 02) equivalent to 1% of the total weight of the fish in the holding tank

ACCLIMATION
- Acclimation period: 24 to 30 h prior starting of test
- Acclimation conditions (same as test or not): same as test
- Type and amount of food during acclimation: no feeding during acclimation
- Mortality during 14-day period prior to transferring a batch of fish to acclimation tank: preliminary test: 2.6%; first definitive test: 2.3%; second definitive test: 0.1%
Test type:
static
Water media type:
other: borehole water
Limit test:
no
Total exposure duration:
96 h
Hardness:
preliminary: 223 - 234 mg/L as CaCO3
first definitive: 200 - 240 mg/L as CaCO3
second definitive: 204 - 230 mg/L as CaCO3
Test temperature:
preliminary: 15.4 - 16.4°C
first definitive: 13.5 - 14.3°C
second definitive: 14.2 - 15.2°C
pH:
preliminary: 8.08 - 8.27
first definitive: 8.16 - 8.35
second definitive: 8.14 - 8.32
Dissolved oxygen:
preliminary: 91 - 98% ASV (air saturation value)
first definitive: 82 - 95% ASV
second definitive: 84 - 91% ASV
Nominal and measured concentrations:
preliminary test:
- nominal: control, 560, 1000 mg/L
first definitive test:
- nominal: control, 560, 1000, 1800 mg/L
second definitive test:
- nominal: control, 560, 1000, 1800, 2400 mg/L
- measured:
Details on test conditions:
TEST SYSTEM
- Test vessel: 14 L all-glass aquaria
- Material, size, headspace, fill volume: 10 L
- Aeration: through Pasteur pipette connected to an oil-free supply of compressed air
- No. of organisms per vessel: preliminary test: 5; definitive tests: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: borehole water
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Photoperiod: 14h light, 10h dark, during second definitive test: each 30 minutes dawn and dusk

TEST CONCENTRATIONS
- A second definitive test was conducted to include a higher concentration.
Reference substance (positive control):
not specified
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: preliminary test
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
1 800 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: first definitive test
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 2 400 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: second definitive test
Details on results:
- Behavioural abnormalities: some fish were overturned or swimming spirally in concentrations down to 560 mg/L until the end of the test (96 h)
- Other biological observations: some fish in all concentrations of the second definitive test showed darker pigmentation and loss of balance when swimming. The number of affected fish increased with concentration.
- Mortality of control: no mortality in any control group
- Other adverse effects control: no adverse effects were observed in the control groups.
Sublethal observations / clinical signs:

preliminary test:

nominal concentration
[mg/L]
number exposed mortality after 96h
[%]
control 10 0
560 10 0
1000 10 10

first definitive test:

test vessel number nominal concentration
[mg/L]
number exposed mortality observations
[number of died fish]
2h 16h 24h 48h 72h 96h
1 control 10 0 0 0 0 0 0
2 control 10 0 0 0 0 0 0
3 560 10 0 0 0 0 0 0
4 560 10 0 0 0 0 0 1
5 1000 10 0 0 0 0 1 3
6 1000 10 0 0 0 1 2 3
7 1800 10 0 0 0 1 4 7
8 1800 10 0 0 0 1 2 3

second definitive test:

test vessel number nominal concentration
[mg/L]
number exposed mortality observations
[number of died fish]
15h 24h 48h 72h 96h
1 control 10 0 0 0 0 0
2 control 10 0 0 0 0 0
3 560 10 0 0 0 0 0
4 560 10 0 0 0 0 0
5 1000 10 0 0 0 0 0
6 1000 10 0 0 0 0 0
7 1800 10 0 0 0 0 0
8 1800 10 0 0 0 0 0
9 2400 10 0 0 0 0 2
10 2400 10 0 0 0 0 2
Validity criteria fulfilled:
yes
Conclusions:
The toxicity of the test item on fish (Rainbow trout) was determined in a GLP study according to OECD 203. The 96-h EC50 was determined to be 1800 mg/L (nominal).

Description of key information

The toxicity of the test item on fish (Rainbow trout) was determined in a GLP study according to OECD 203. The 96-h EC50 was determined to be 1800 mg/L (nominal).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Effect concentration:
1 800 mg/L

Additional information

The toxicity of test item on the fish Oncorhynchus mykiss (Rainbow trout) was determined in a GLP study according to OECD 203 covering 2 definitive tests. Individuals of Rainbow trout exposed to 0 (control), 560, 1000, 1800 and 2400 mg/L of the test substance in chalk borehole water, equivalent to geometric mean measured concentrations of <LOQ, 543,998, 1778 and 2414 mg/L during the incubation of 96 h. Mortality of Rainbow trout was observed after 48 h at 1000 mg/L in the first definitive test and after 96 h at 2400 mg/L in the second definitive test. The 96 h EC50 value of >1800 mg/L (nominal) was concluded.