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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:
2020
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Version / remarks:
30 May 2008
Deviations:
yes
Remarks:
On day 4 observations were performed only once. Because no mortality was ob-served after 96 hours, this was stated as uncritical.
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Version / remarks:
Second edition, adopted 08. Feb. 2019.
Deviations:
yes
Remarks:
On day 4 observations were performed only once. Because no mortality was ob-served after 96 hours, this was stated as uncritical.
GLP compliance:
yes (incl. QA statement)
Analytical monitoring:
yes
Details on sampling:
A stock solution containing 1000.3 mg/L in tap water was prepared. The concentration to be tested was prepared by diluting of this stock solution with tap water.
Vehicle:
yes
Details on test solutions:
A stock solution containing 1000.3 mg/L in tap water was prepared. The concentration to be tested was prepared by diluting of this stock solution with tap water.

Test Medium
Good quality drinking water is used. The composition is analysed yearly by the supplier.
Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
Specification
Species: Danio rerio HAMILTON-BUCHANAN
Age: sexually immature young fish, length 2 +/- 1 cm

Origin
The original animals are obtained in 2019 from the University Heidelberg and used for breeding and production of eggs. The offspring of the original animals is used in the test.

Husbandry
Danio rerio is routinely used for toxicity tests. The test fish are kept under following conditions:
Vessels: polyethylene aquaria
Medium: tap water
Feeding: three times a day with warmwater fishfood
Photo period: 12/12 hours, using neon tubes
Temperature: 23 +/- 2 °C
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h
Post exposure observation period:
Observations were made after 1 h, 4 h and 24 h after the start of the exposure (day 0-1). On days 2-3 of the test, all vessels with were inspected twice per day (morning and afternoon). On day 4, the vessels were inspected only at the end of the experiment.
The content of the test item in the test vessels was measured at the beginning and at the end of the study.
Hardness:
Blank control hardness in mg CaCO3/L = 106.8
Drinking water hardness = 0.88 mmol/L
Test temperature:
23 ± 2 °C
pH:
Blank control: 7.9 (0 h); 7.8 (24 h); 7.8 (48 h); 7.8 (72 h); 7.9 (96 h)
solution 100 mg/L: 7.2 (0 h); 7.7 (24 h); 7.8 (48 h); 7.8 (72 h); 7.9 (96 h)
Dissolved oxygen:
Blank control [O2 in mg/L]: 8.9 (0 h); 8.2 (24 h); 6.8 (48 h); 7.4 (72 h); 7.1 (96 h)
solution 100 mg/L [O2 in mg/L]: 8.2 (0 h); 8.5 (24 h); 7.6 (48 h); 7.8 (72 h); 7.5 (96 h)
Salinity:
sodium: 4.4 mg/L
potassium: 2.5 mg/L
calcium: 32 mg/L
magnesium: 2.1 mg/L
aluminium: 0.007 mg/L
iron: <0.005 mg/L
manganese: <0.005 mg/L
ammonium: <0.05 mg/L
nitrate: 1.4 mg/L
nitrite: <0.01 mg/L
chloride: 9.2 mg/L
sulphate: 10 mg/L
Conductivity:
Conductivity of drinking water at 25 °C = 210 µS/cm
Nominal and measured concentrations:
At the start and at the end of the test, the content of the test item in the test solutions was determined using HPLC. The measured concentrations lay between 99 % and 109 % of the nominal concentrations. Therefore, the determination of the results was based on the nominal concentrations (see OECD Guidance Doc. No.23 (§176)).
Details on test conditions:
24 hours before the start of the test, the test fish were no longer fed.
Date: 11. – 15. Nov. 2019
Treatment: 100 mg/L
Test Design: static
Medium renewal: none
Duration: 96 hours
Loading: 1 fish/L
Vessels: glass aquaria, maximal volume 12 L
Aeration: accomplished with glass tubes
Feeding: none
Photo period: 12/12 hours using neon tubes
Temperature: 23 ± 2 °C
pH adjustment: none
Replicate (Treatments): 1 vessel, each containing 7 L test solution and 7 fish
Replicate (Blank Control): 1 vessel, containing 7 L tap water and 7 fish

Observations were made after 1 h, 4 h and 24 h after the start of the exposure (day 0-1). On days 2-3 of the test, all vessels with were inspected twice per day (morning and afternoon). On day 4, the vessels were inspected only at the end of the experiment.

The content of the test item in the test vessels was measured at the beginning and at the end of the study.
Reference substance (positive control):
no
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: the tested concentration showed no mortality.
Details on results:
Please see detials on results reported in section "any other information on results incl. tables".
Results with reference substance (positive control):
Positive control has not been used.
Reported statistics and error estimates:
The tested concentration showed no mortality. Therefore, no statistical evaluation of the biological results was performed and the LC50 is given as a range.
Sublethal observations / clinical signs:

The mortalities were recorded daily and can be found in the following table.

Table. Mortality (1 h – 24 h)

Nominal Conc. in mg/L

Total number of fish used

Number of dead fish

1 h

4 h

24 h

Blank control

7

0

0

0

100

7

0

0

0

 

Table. Mortality (day 2 – 4)

Nominal Conc. in mg/L

Total number of fish used

Number of dead fish

2 d

3 d

4 d

morning

afternoon

morning

afternoon

Blank control

7

0

0

0

0

0

100

7

0

0

0

0

0

 

No mortality and no abnormal behaviour of the surviving fish was observed.

Validity criteria fulfilled:
yes
Conclusions:
The tested concentration showed no mortality; therefore, 96h LC50 for fish is > 100 mg/L.
Executive summary:

One valid experiment was performed.

The toxicity against Danio rerio was tested using a static test design according to OECD Guideline 203. The control and the tested concentration showed no mortality. No abnormal behaviour of the surviving fish was observed.

At the start and at the end of the test, the content of the test item in the test solutions was determined using HPLC. The measured concentrations lay between 99 % and 109 % of the nominal concentrations. Therefore, the determination of the results was based on the nominal concentrations (see OECD Guidance Doc. No.23 (§176)).

No observations were made which might cause doubts concerning the validity of the study outcome. All validity criteria were met.

The result of the test is considered valid.

Description of key information

The tested concentration showed no mortality; therefore, 96h LC50 for fish is > 100 mg/L.

One valid experiment was performed.

The toxicity against Danio rerio was tested using a static test designaccording to OECD Guideline 203. The control and the tested concentration showed no mortality. No abnormal behaviour of the surviving fish was observed.

At the start and at the end of the test, the content of the test item in the test solutions was determined using HPLC. The measured concentrations lay between 99 % and 109 % of the nominal concentrations. Therefore, the determination of the results was based on the nominal concentrations (see OECD Guidance Doc. No.23 (§176)).

No observations were made which might cause doubts concerning the validity of the study outcome. All validity criteria were met.

The result of the test is considered valid.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Effect concentration:
100 mg/L

Additional information

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