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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:
2004-07-05 to 2004-11-01
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Qualifier:
according to guideline
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Qualifier:
according to guideline
Guideline:
other: ISO 7346-2: semi-static method
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
- Concentrations: 0, 1, 3.2 and 10 mg/L (nominal)
- Sampling method: 0, 72 h (fresh), 24 and 96 h (old), 10 mL from approximate centre of vessels
- Sample storage conditions before analysis: Not applicable, samples analysed on day of sampling.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Test concentrations prepared daily from 100 mg/L stock solution. 10-20 minutes magnetic stirring used to accelerate disolving of the test substance in the test media. Layer of foam dissolved in test medium by use of air pressure.
- Controls: Blank
- Evidence of undissolved material (e.g. precipitate, surface film, etc): No
Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: Zebra-fish
- Strain: Not specified
- Source: Gommers-Ducheine BV, Gendt, The Netherlands
- Age at study initiation (mean and range, SD): Not specified
- Length at study initiation (length definition, mean, range and SD):
1st range finding: 3.2 ± 0.2 cm
2nd range finding: 3.2 ± 0.1 cm
Final test: 3.2 ± 0.2 cm
- Weight at study initiation (mean and range, SD):
1st range finding: 0.55± 0.12 g
2nd range finding: 0.61± 0.19 g
Final test: 0.61 ± 0.15 g
- Feeding during test: no, from 48 h prior to test and throughout test period.

ACCLIMATION
- Acclimation period: 12 days
- Acclimation conditions (same as test or not): Yes
- Type and amount of food: Trouvit, amount not specified
- Feeding frequency: daily
- Health during acclimation (any mortality observed): During 7 days prior to start of test mortality was less than 5%

Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Post exposure observation period:
None
Hardness:
250 mg CaCO3/L
Test temperature:
21.3 - 22.0°C
pH:
7.9 - 8.0 mg/L
Dissolved oxygen:
6.1 - 9.0
Salinity:
Not applicable
Nominal and measured concentrations:
Final:
nominal: 1.0, 1.8, 3.2, 5.6 and 10 mg/L (see attached document: 04-015B Exposure concentrations)
Details on test conditions:
TEST SYSTEM
- Test vessel: 10.5 L glass
- Material, size, headspace, fill volume: 10L test medium
- Aeration: No
- Type of flow-through (e.g. peristaltic or proportional diluter): No data
- Renewal rate of test solution (frequency/flow rate): every 24 hours
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): one
- No. of vessels per control (replicates): one
- Biomass loading rate: 0.43 g fish/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: tap water purified by reverse osmosis
- Culture medium different from test medium: No

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 h light
- Light intensity: Not specified

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :

TEST CONCENTRATIONS
- Spacing factor for test concentrations: =< 2.2
- Range finding study: yes, 2
- Test concentrations:
1st range finder: 0.1 - 100 mg/L increasing by a factor of 10
2nd range finder: 0.1 and 1.0 mg/L
- Results used to determine the conditions for the definitive study: Yes
Reference substance (positive control):
yes
Remarks:
pentachlorophenol (PCP) - periodical check on sensitivity of test system
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
4.2 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
mortality (fish)
Remarks on result:
other: 95% CL = 3.2 - 5.6 mg/L Reached by 48 h exposure
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
3.2 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
other: clinical effects
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
5.6 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
mortality (fish)
Remarks on result:
other: Reached by 48 h exposure
Details on results:
Hypoactivity was noted in all 7 fish dosed with 10 mg/L at the 2h observation and in 2/3 fish dosed with 5.6 mg/L at the 24 h observation.
Results with reference substance (positive control):
- Results with reference substance valid? Yes
- Mortality: LC100 = 0.22 mg/L
- LC50: 0.11 mg/L (48h)
Historical data on LC50 for the test facility is in range 0.10 - 0.46 mg/L
- Other:
Sublethal observations / clinical signs:

Table 1: Incidence of mortality and total mortality (final test)

Nominal conc. (mg/L)

Initial no. fish

Cumulative mortality

Total mortality (%)

2h

24h

48h

72h

96h

Blank control

7

0

0

1*

1

1

14

1.0

7

0

0

0

0

0

0

1.8

7

0

0

0

0

0

0

3.2

7

0

0

0

0

0

0

5.6

7

0

4

7

7

7

100

10

7

0

7

7

7

7

100

*Fish jumped out of test vessel

Validity criteria fulfilled:
yes
Conclusions:
Under the conditions of the study, the test material produced no visble effects in zebra-fish at or below 3.2 mg/L (NOEC)
The 96 h LC50 was 4.2 mg/L based on analytically confirmed nominal exposure concentrations (with 0% mortality at 3.2 mg/L and 100% mortality at 5.6 mg/L) and was already reached after 48 hours of exposure.
Executive summary:

In a semi-static 96-hour acute toxicity study in zebra-fish performed in accordance with OECD TG 203 and EU Method C.1 the test material was dosed at nominal concentrations of 1.0, 1.8, 3.2, 5.6 and 10 mg/L to 7 fish per dose. The dose concentrations were chosen using two range finding studies. A blank control was run in conjunction with the final test. Results of a positive control study on pentachlorophenol (PCP) were appended to the study to confirm sensitivity of the test system.

Analysis of samples taken at the start and after 72 hours of exposure of the freshly prepared test solutions showed that measured concentrations were in agreement with nominal. The 10 mg/L solution remained stable in the 24 hours before renewal of the test solutions. At nominal 3.2 mg/L the test concentration remained fairly stable, but at nominal 1.0 mg/L actual test concentrations generally decreased significantly. The reason is unknown.

The biological results showed that the nominal test concentrations of 1.0 and 1.8 mg/L were toxicologically not relevant for determination of an LC50 value. As nominal concentrations of 3.2 and 10 mg/L were in agreement it was expected that the actual concentration at 5.6 mg/L would also be in agreement with nominal. Therefore LC50 and NOEC values were based on nominal concentrations.

The study met the acceptability criteria and was considered valid.

Under the conditions of the study, the test material produced no visble effects in zebra-fish at or below 3.2 mg/L (NOEC).

The 96 h LC50 was 4.2 mg/L (95% CL = 3.2 - 5.6 mg/L) based on analytically confirmed nominal exposure concentrations (with 0% mortality at 3.2 mg/L and 100% mortality at 5.6 mg/L) and was already reached after 48 hours of exposure.

Description of key information

The LC50 (96-h) for fish is 4.2 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Effect concentration:
4.2 mg/L

Additional information

One reliable study is available. In this study performed under GLP according to OECD TG 203 groups of seven Danio rerio (Zebra fish) were exposed to the substance under semi-static conditions for 96 hours at nominal concentrations of 0, 1.0, 1.8, 3.2, 5.6 or 10 mg/L [Bouwman LM (2004)]. The 96 -h LC50 was 4.2 mg/L based on nominal test concentrations.