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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:
2016-08-23 till 2017-01-11
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
Concentration control analyses were not performed, because a sufficiently sensitive method to determine the test substance in test water up to its saturation limit was not available.
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
yes
Remarks:
Concentration control analyses were not performed
Qualifier:
according to guideline
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Version / remarks:
2008
Deviations:
yes
Remarks:
Concentration control analyses were not performed
GLP compliance:
yes (incl. QA statement)
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
The test substance is a mixture of sparingly water soluble components. Therefore the test solutions were prepared following general guidance provided in OECD 23 in order to achieve a water accommodated fraction (WAF) of the test substance using a liquid-liquid saturator technique. For each test solution, a glass aspirator bottle (approx. 20 L) with a bottom side-outlet attached to a stopcock was filled with 20 L test media. Prior to use the test substance was homogenized by shaking of the container. Then 73.5 μL (considering density = 0.8166 g/cm³) of the test substance was pipetted carefully on the water surface. The bottle was closed and the solution was stirred on a magnetic plate for about 3 days. Undissolved test substance was observed at the water surface as oily drops and the aqueous phase of the stock solution was clear. The required volume of the WAF (approx. 10 L per test solution) was removed from the bottom port. The exposure was started after separation of the undissolved material. The control was treated in the same way, but in 10 L flasks. All test solutions were visibly clear following preparation. Fresh test solutions were prepared daily. The test vessels were conditioned with the test solution for approx. 72-h prior initiating exposure.
Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Source: Kölle Zoo, Ludwigshafen/Rhein, Germany
- Age at study initiation: Approx. 9 months
- Hatching date: Dec 2015
- Arrival at the testing facility: 25 May 2016


ACCLIMATION
- Acclimation period: yes, 14 days
- Acclimation conditions: same as test conditions
- Type and amount of food during acclimation: ”Tetramin“ (manufacturer Tetra-Werke, Melle, Germany) daily ad libitum, additionally generally on workdays live juvenile brine shrimp (Artemia nauplii).
- Health during acclimation: no mortality, no medical treatment
Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h
Hardness:
100 mg/L CaCO3
Test temperature:
23 ±1°C
pH:
8.0 - 8.3
Dissolved oxygen:
8.2 - 8.4 mg/L, >60%
Nominal and measured concentrations:
0 - 3 mg/L nominal concentrations
Details on test conditions:
TEST SYSTEM
- Test vessel: Approx. 10 Liter stainless steel aquaria (29x21x22 cm)
- Fill volume: 10 L
- Aeration: Continuous slight aeration via glass capillary
- Renewal rate of test solution: The renewal period was 24 hours
- No. of organisms per vessel: 7 fish/ test vessel
- No. of vessels per concentration: 1 replicate
- No. of vessels per control: 1 replicate
- Biomass loading rate: 0.25 g fish/L

TEST MEDIUM / WATER PARAMETERS
- Preparation of dilution water: non chlorinated charcoal-filtered drinking water from the municipal water works of the city of Frankenthal mixed with deionized water
- Hardness: 100 mg/L as CaCO3
- pH: 7.5 - 8.5

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16h light, 8h dark
- Light intensity: 114–431 Lux

EFFECT PARAMETERS MEASURED:
- Fish: mortality and toxic signs (at hour 1, 6, 24, 48, 72, 96)

TEST CONCENTRATIONS
In the interest of animal welfare, the concentrations for this study were selected based on the recommendations in OECD 126 on a threshold approach for acute fish toxicity. The value derived from the results of algal growth study with the test substance (i.e. the threshold concentration). If fish toxicity is above this level, a further fish test is not required since the environmental assessment will be based on the more sensitive species.
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
>= 3 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other:
Remarks:
threshold approach
Duration:
96 h
Dose descriptor:
LC0
Effect conc.:
>= 3 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other:
Remarks:
threshold approach
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
> 3 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other:
Remarks:
threshold approach
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
> 3 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other:
Remarks:
threshold approach
Details on results:
- Behavioural abnormalities: no
- Observations on body length and weight: lenght (3.0 cm – 3.9 cm, mean 3.5 cm), weight (0.20 g – 0.48 g, mean 0.35g)
- Mortality of control: no
- Other adverse effects control: no
- Abnormal responses: no
Reported statistics and error estimates:
Since the study is designed as a limit test, the data were not appropriate for the use of computerized methods for the calculation of point estimates. Any reported point estimates (NOEC and LCx values) are estimated based on expert judgment in accordance with test guideline recommendations.
Sublethal observations / clinical signs:

Table 1: Mortality of Fish

Loading rate [mg/L]

Inihital number of fish

Cumulative mortality (number of dead animals)

1 h

6 h

24 h

48 h

72 h

96 h

0

7

0

0

0

0

0

0

3

7

0

0

0

0

0

0

Table 2: Toxic signs of Fish

Loading rate [mg/L]

Inihital number of fish

Observed signs of toxicity

1 h

6 h

24 h

48 h

72 h

96 h

0

7

n

n

n

n

n

n

3

7

n

n

n

n

n

n

n: no toxic signs observed

Validity criteria fulfilled:
yes
Conclusions:
In this study the test item was found to cause no lethal effects to zebrafish after 96 hours at a nominal concentration of 3 mg test item/L.
Executive summary:

The acute toxicity of the test item to fish (zebrafish) was determined according to the principles of the OECD-Guideline for Testing of Chemicals No. 203 (1992) and EU Council Regulation (EC) No. 440/2008/ Method C.1 (2008) from 2016-08-29 to 2016-09-02. An acute toxicity test under semi-static conditions with renewal of the test media each 24 hours was conducted with the nominal test item concentration of 0 and 3 mg/L. Duration of the test was 96 hours. Seven test organisms were exposed to each test concentration and the control. Water quality parameters (temperature, pH-value and oxygen- saturation) measured at 0, 24, 48, 72 and 96 hours were determined to be within the acceptable limits. All validity criteria of the test guideline were met. Since the study is designed as a limit test, the data were not appropriate for the use of computerized methods for the calculation of point estimates. Any reported point estimates (NOEC and LCx values) are estimated based on expert judgment in accordance with test guideline recommendations. In this study the test item was found to cause no lethal effects to zebrafish after 96 hours at a nominal concentration of 3 mg test item/L, so the NOEC and LC100 are >3 mg/L, the LC50 and LC0 are >=3 mg/L.

Description of key information

Key study: In this study according to OECD Guideline Test No. 203 the test item was found to cause no lethal effects to zebrafish after 96 hours at a nominal concentration of 3 mg test item/L, so the LC 50 is above 3 mg/L.

Key value for chemical safety assessment

Additional information

Key study: The acute toxicity of the test item to fish (zebrafish) was determined according to the principles of the OECD-Guideline for Testing of Chemicals No. 203 (1992) and EU Council Regulation (EC) No. 440/2008/ Method C.1 (2008) from 2016-08-29 to 2016-09-02. An acute toxicity test under semi-static conditions with renewal of the test media each 24 hours was conducted with the nominal test item concentration of 0 and 3 mg/L. Duration of the test was 96 hours. Seven test organisms were exposed to each test concentration and the control. Water quality parameters (temperature, pH-value and oxygen- saturation) measured at 0, 24, 48, 72 and 96 hours were determined to be within the acceptable limits. All validity criteria of the test guideline were met. Since the study is designed as a limit test, the data were not appropriate for the use of computerized methods for the calculation of point estimates. Any reported point estimates (NOEC and LCx values) are estimated based on expert judgment in accordance with test guideline recommendations. In this study the test item was found to cause no lethal effects to zebrafish after 96 hours at a nominal concentration of 3 mg test item/L,so the NOEC and LC100 are >3 mg/L, the LC50 and LC0 are >=3 mg/L.