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Ecotoxicological information

Short-term toxicity to fish

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Link to relevant study record(s)

Reference
Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
March 14-18, 2005
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
- Concentrations: 3,6,1,24,48 mg/L (nominal)
- Sampling method: direct
- Sample storage conditions before analysis: same as test conditions
Vehicle:
no
Details on test solutions:
The test media (reconstituted water and test material) were freshly prepared. Therefore, the calibrated flask with test material and vehicle, reconstituted water, was treated in an ultrasonic device for 1 hour. Subsequently, the preparation was stirred with a magnetic stirrer for further 23 hours. After 24 hours the preparation was given for 6 hours through a separatory funnel. The stock preparation was diluted with reconstituted water to the different test media concentrations.
Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
Species: zebra fish (Danio rerio), originated West-Aquarium
Breeder: in house
Size: 2.0 + 1.0 cm
Body weight per group: 3:5 and 5:5 g

The species is internationally accepted for this type of study.
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Test temperature:
23 to 26 °C
pH:
7.4 - 8.1
Dissolved oxygen:
> 60 % saturation
Nominal and measured concentrations:
nominal: 3, 6, 12, 24, and 48 mg/L
measured: 0.063, 0.149, 0.266, 0.579, 1.105 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: 9L
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: headspace
- Aeration: yes, for 24 hours before start of exposure
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 1

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 12/12 hours
- Light intensity: no data

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : clinical behavior, mortality
Duration:
24 h
Dose descriptor:
LC50
Effect conc.:
>= 1.1 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
>= 1.1 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
72 h
Dose descriptor:
LC50
Effect conc.:
0.8 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: 0.6-1.0
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
0.8 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: 0.6-1.0
Key result
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
0.063 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Details on results:
No signs of toxicity were seen in the control group and in the zebra fish exposed to nominal concentrations of 3 mg/L.
Signs of toxicity were seen in the nominal concentrations 6 to 48 mg/L. They started in the higher concentrations after 48 hours and lasted up to the end of the experimental part. These consisted of tumbling during swimming, swimming behavior slightly irregular, fish at the water surface, and fish near the bottom of the aquarium.
Deaths occurred only in the highest concentration, after an exposure period of about 72 to 96 hours.
Validity criteria fulfilled:
yes
Conclusions:
Under the given conditions of this study the 96 h LC50 value for fish was 0.8 mg/L and the NOEC was 0.063 mg/L.
Executive summary:

Study Design

The objective of this study was to determine the acute fish toxicity of the test material using zebra fish (Dania rerio). There was a very poor recovery of the test material after 48 hours and therefore a closed semistatic test system was chosen. The test material was renewed in 24 hours intervals. For this purpose, 7 zebra fish per test material group and further 7 zebra fish in a control group were exposed over 96 hours, under defined conditions. The fish were observed for signs of toxicity or death. Zebra fish (Danio rerio) were exposed to aqueous test material solutions of nominal 3, 6, 12, 24, and 48 mg/L in a closed semistatic system.

Results

The limit of quantification of the analytical method was 0.016 mg/L. The analytically determined concentrations were between 2 - 4 % at the start and 1 - 3 % at the end of the 24 hours exposure period. During the experimental phase of the study, the test material concentrations could not be maintained at >= 80 % of the initial concentrations. Therefore, the LC50 values had to be calculated with the mean analytical concentrations.
No signs of toxicity were seen in the control group and in the zebra fish exposed to nominal concentrations of 3 mg/L. Signs of toxicity were seen in the nominal concentrations 6 to 48 mg/L. They started in the higher concentrations after 48 hours and lasted up to the end of the experimental part. These consisted of tumbling during swimming, swimming behavior slightly irregular, fish at the water surface, and fish near the bottom of the aquarium. Deaths occurred at highest concentration, after an exposure period of about 72 to 96 hours.
Nominal and analytical concentrations, and number of deaths per time point were given below:

Concentration [mg/L]

Dead / exposed fish
Nominal
Mean analytical
24h
48h
72h
96h
0, control
0
0/7
0/7 0/7 0/7
3
0.063
0/7 0/7 0/7 0/7
6
0.149
0/7 0/7 0/7 0/7
12
0.266
0/7 0/7 0/7 0/7
24
0.579
0/7 0/7 0/7 0/7
48
1.105
0/7 0/7 7/7
7/7


For the test material, the following analytical LC50 values and no effect concentration for zebra fish (Dania rerio) were determined:
24 h LC50 > 1.1 mg/L
48 h LC50 > 1.1 mg/L
72 h LC50 = 0.8 (0.6-1.0) mg/L
96 h LC50 = 0.8 (0.6-1.0) mg/L
NOEC = 0.063 mg/L

Conclusion

Under the given conditions of this study the 96 h LC50 value for fish was 0.8 mg/L and the NOEC was 0.063 mg/L.


Description of key information

Under the given conditions of this study the 96 h LC50 value for fish was 0.8 mg/L and the NOEC was 0.063 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Effect concentration:
0.8 mg/L

Additional information