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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:
from 2014-04-11 to 2014-10-22
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Qualifier:
according to guideline
Guideline:
other: Guidelines for the Testing of Chemical, SEPA, P.R. China No. 203 " Fish, Acute Toxicity Test" (2004)
Deviations:
no
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
yes
Remarks:
gas chromatography
Details on sampling:
The concentrations of the solutions were tested at the begining and at the end of the test.
Vehicle:
no
Details on test solutions:
WAF solutions were prepared by adding 0.0253 g, 0.0355 g, 0.0507 g, 0.0709 g and 0.1012 g of test item to 10L test medium respectively and adding 0.0759 g, 0.1063 g, 0.1517 g, 0.2124 g and 0.3033 g of test item to 30L test medium respectively, and the solutions with loading rate of 2.5 mg/L, 3.5 mg/L, 5 mg/L, 7 mg/L and 10 mg/L were obtained. The solutions were mixed for 48 h at the speed of 300 rpm using electric stirrer at 30 ± 2 °C. Then the 10L and 30L solutions were combined after mixing and a total of 40L solutions with chosen loading rate were obtained. Following cessation of mixing and settling for 4 hours, the WAFs were filtered with glass wool and the final test solutions were obtained.
Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: Zebra fish
- Length at study initiation: 2.1 to 3.0 cm

ACCLIMATION
- Acclimation period: 170 days
- Acclimation conditions: same as test
- Feeding frequency during acclimation: once a day, ceasing 24 hours before the start of the test
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Hardness:
10 to 250 mg CaCO3 per L
Test temperature:
23 ± 2 °C
pH:
8.28 - 8.31
Dissolved oxygen:
5.27 - 8.58 mg/L
Nominal and measured concentrations:
Nominal concentrations: 2.5, 3.5, 5, 7 and 10 mg/L
Measured concentrations: 1.61, 2.15, 3.61, 4.67 and 5.52 mg/L
Details on test conditions:
TEST SYSTEM
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 3
- Biomass loading rate: 0.25 g fish/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: tap water treated with an active carbon water filter, the quartz sand water filter, micro-filtration water filter, ultraviolet light and aeration

OTHER TEST CONDITIONS
- Photoperiod: 12 hours light photopcriod daily
- Light intensity: lamplight and natural light

OBSERVATIONS
- at 3, 6, 24, 48, 72 and 96 hours
Reference substance (positive control):
yes
Remarks:
Potassium dichromate
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
4.1 mg/L
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Details on results:
- Nomortality in the blank control was observed.
- Highest concentration producing 0 % mortality during test was 2.15 mg/L based on measured concentration.
- Lowest concentration producing 100 % mortality during test was 5.52 mg/L based on measured concentration.
Results with reference substance (positive control):
The 24h LC50 value of Potassium dichromate was determined to be 248.542 mg/L.
Sublethal observations / clinical signs:

Table 1. Toxicity symptoms and mortality (Preliminary test)

Concentration (mg/L)

Fish Number

Time (hr)

Cumulative Mortality

Abnormalities

Blank control

5

24

0

None

48

0

72

0

96

0

1.25

5

24

0

None

48

0

72

0

96

0

2.5

5

24

0

None

48

0

72

0

96

0

5

5

24

0

Death, loss of equilibrium, swimming behavior decreased

48

0

72

1

96

1

10

5

24

0

Death, loss of equilibrium

48

0

72

4

96

5

20

5

24

0

Death, loss of equilibrium

48

4

72

5

96

5

Table 2. Cumulative Mortality at Each Concentration

Concentration (mg/L)

(loading rate/measured)

Cumulative mortality (%)

24 h

48 h

72 h

96 h

2.5/1.61

0 %

0 %

0 %

0 %

3.5/2.15

0 %

0 %

0 %

0 %

5/3.61

0 %

13 %

17 %

30 %

7/4.67

0 %

33 %

33 %

63 %

10/5.52

0 %

70 %

93 %

100 %

The acute toxicity of the test item was calculated in terms of measured concentrations and the results were as follows:

24-h LC50: > 5.52 mg/L

48-h LC50: 4.968 mg/L with 95 % confidence interval of 4.617 - 5.504 mg/L

72-h LC50: 4.604 mg/L with 95 % confidence interval of 2.645 - 9.898 mg/L

96-h LC50: 4.100 mg/L with 95 % confidence interval of 3.813 - 4.348 mg/L

Table 3. Toxicity symptoms and mortality(Maintest)

Nominal Concentration (mg/L)

Fish Number

Time (hr)

Cumulative Mortality

Abnormalities

Replicate

Average

1

2

3

Blank control

30

3

0

0

0

0

None

6

0

0

0

0

24

0

0

0

0

48

0

0

0

0

72

0

0

0

0

96

0

0

0

0

2.5 mg/L

30

3

0

0

0

0

None

6

0

0

0

0

24

0

0

0

0

48

0

0

0

0

72

0

0

0

0

96

0

0

0

0

3.5 mg/L

30

3

0

0

0

0

None

6

0

0

0

0

24

0

0

0

0

48

0

0

0

0

72

0

0

0

0

96

0

0

0

0

5 mg/L

30

3

0

0

0

0

Death, loss of equilibrium, swimming behavior decreased

6

0

0

0

0

24

0

0

0

0

48

2

1

1

1.3

72

3

1

1

1.7

96

4

2

3

3

7 mg/L

30

3

0

0

0

0

Death, loss of equilibrium, swimming behavior decreased

6

0

0

0

0

24

0

0

0

0

48

4

3

3

3.3

72

4

3

3

3.3

96

7

5

7

6.3

10mg/L

30

3

0

0

0

0

Death, loss of equilibrium, swimming behavior decreased

6

0

0

0

0

24

0

0

0

0

48

7

8

6

7

72

10

9

9

9.3

96

10

10

10

10

 

Analytical results

According to the results of the stability test, the concentration of the test item in water could maintain within 80 - 120 % of initial. So the test was conducted with static regime and the test vessel was covered during the test.

Table 4. Concentration measurement in the main test

Nominal concentration (mg/L)

Replicate

Measured concentration 0 h (mg/L)

Measured concentration 96 h (mg/L)

2.5

1

1.57

1.88

2

1.63

1.62

3

1.63

1.45

3.5

1

2.17

2.23

2

2.13

1.86

3

2.16

1.94

5

1

3.53

3.68

2

3.61

3.00

3

3.69

3.02

7

1

4.64

4.06

2

4.73

4.42

3

4.65

4.12

10

I

5.34

5.69

2

5.78

5.86

3

5.43

5.49

 

Validity criteria fulfilled:
yes
Conclusions:
The toxicity of the test item on Zebrafish was determined in a 96-h study. The LC50 was determined to be 4.1 mg/L.
Executive summary:

The acute toxicity of the test item to fish (zebrafish) was determined according to The Guidelines for the Testing of Chemical, SEPA, P.R. China No. 203 "Fish, Acute Toxicity Test" (2004)and OECD Guideline 203. A static test was performed with nominal concentrations of 2.5 mg/L, 3.5 mg/L, 5 mg/L, 7 mg/L and 10 mg/L. The test item solution was prepared by WAF method in filtered tap water. 30 test organisms in 3 replicates were exposed to the test item solutions and the control each for 96 hours. The measured concentration of the test item at the start and at the end of the exposure period was in the range 80 - 120 %. Therefore, the biological endpoints were based on the measured concentrations. The LC50 after 96 h was determined to be 4.1 mg/L.

Description of key information

The toxicity of the test item on Zebra fish was determined in a 96-h study. The LC50 was determined to be 4.1 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Effect concentration:
4.1 mg/L

Additional information

The acute toxicity of the test item to fish (zebra fish) was determined according to The Guidelines for the Testing of Chemical, SEPA, P.R. China No. 203 "Fish, Acute Toxicity Test" (2004) and OECD Guideline 203. A static test was performed with nominal concentrations of 2.5 mg/L, 3.5 mg/L, 5 mg/L, 7 mg/Land 10 mg/L. The test item solution was prepared by WAF method in filtered tap water. 30 test organisms in 3 replicates were exposed to the test item solutions and the control each for 96 hours. The measured concentration of the test item at the start and at the end of the exposure period was in the range 80 - 120 %. Therefore, the biological endpoints were based on the measured concentrations. The LC50 after 96 h was determined to be 4.1 mg/L.