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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
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Experimental test result performed according to the guideline.
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Principles of method if other than guideline:
Study was conducted to access the effect of test chemical on the growth of fish Danio rerio. Test conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test).
GLP compliance:
not specified
Analytical monitoring:
yes
Vehicle:
not specified
Details on test solutions:
The test substance was partially soluble in water. The in house water solubility in DI water was 21.6523 mg/L.Therefore, the stock solution was prepared by dissolving 5 g of the test substance in 5 liters of potable water (passed through reverse osmosis system) with continuous 72 hrs stirring. After stirring the stock analytically detected and solubility in RO water found to be 122.575 mg/L.By using this value of solubility following test concentrations were prepared 6.25mg/L,12.5mg/L,25mg/L,50mg/L,100mg/L ,respectively.
Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: Zebra Fish
- Strain: Danio rerio
- Source: Anytime Pet,Jaripatka, Nagpur
- Length at study initiation (length definition, mean, range and SD): average 1.8 cm
- Weight at study initiation (mean and range, SD): average 0.082 g
-Housing:The fishes were kept in a static tank in tap water passed through reverse osmosis system, under natural conditions along with proper feed and aeration.
-Duration of housing: 15 days
-Photoperiod: 16 hours light and 8 hours dark
-Diet : Standard brand feed, once daily
-Acclimatization: seven days
-feeding frequency: Once in a day
Test type:
static
Water media type:
freshwater
Total exposure duration:
96 h
Post exposure observation period:
24, 48, 72 and 96 hours
Hardness:
155 mg of CaCO3
Test temperature:
23 °C
pH:
7.2 to 7.4
Dissolved oxygen:
7.0 mg/l
Nominal and measured concentrations:
6.25 mg/L,12.5 mg/L, 25 mg/L, 50 mg/L and 100 mg/L.
Details on test conditions:
TEST SYSTEM
- Test vessel: Plastic aquaria
- Material, size, headspace, fill volume: 5 liters of plastic aquaria filled with 2 liter of water
- Aeration: Aeration in test vessels was provided 1 day before the start of experiment
- No. of organisms per vessel: 8 fishes

OTHER TEST CONDITIONS
- Photoperiod: 16 hours light and 8 hours dark

Reference substance (positive control):
not specified
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
6.25 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: Other details not known
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
12.5 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: Mortality were observed

1. Mortality and symptoms:

Nominal

Concentration

(mg/l)

Number of

fish

Mortality (cumulated) after

 

 

 

3hr

6hr

24hr

48hr

72hr

96hr

 

 

 

V1

V2

V1

V2

V1

V2

V1

V2

V1

V2

V1

V2

Control

4

4

-

-

-

-

-

-

-

-

-

-

-

-

6.25

4

4

-

-

-

-

-

-

-

-

1

1

2

2

12.5

4

4

-

-

-

-

4

4

4

4

4

4

4

4

25

4

4

-

-

-

-

4

4

4

4

4

4

4

4

50

4

4

-

-

4

4

4

4

4

4

4

4

4

4

100

4

4

-

-

4

4

4

4

4

4

4

4

4

4

2. Visible Symptoms:

Nominal

Concentration

(mg/l)

Number of

fish

symptom (cumulated) after

 

 

 

3hr

6hr

24hr

48hr

72hr

96hr

 

4

4

V1

V2

V1

V2

V1

V2

V1

V2

V1

V2

V1

V2

Control

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

6.25

4

4

NS4

NS4

M4

M4

M4

M4

M4

M4

M3

M3

M2

M2

12.5

4

4

NS4

NS4

M4

M4

-

-

-

-

-

-

-

-

25

4

4

NS4

NS4

M4

M4

-

-

-

-

-

-

-

-

50

4

4

M4

M4

-

-

-

-

-

-

-

-

-

-

100

4

4

M4

M4

-

-

-

-

-

-

-

-

-

-

Explanation of symptoms:

A: apathy

N: Narcotic-like state

C: Swimming near water surface

M: Slow movements compared to control

NS: No abnormal symptoms

S: Surfacing behaviour

L: Loss of equilibrium

SM: Shimming movement

H1: Hyperactivity

H2: Hypo activity

B: Bending behavior

F: Fin rot

IS: Invert Swimming

SB: Swimming behaviour

RF: Respiratory Function

P: Pigmentation

Fig 3: pH and oxygen content and temperature:

 

 

Number of

pH readings after

 

 

 

 

Nominal

fish

24h

 

48h

 

72h

 

96h

 

Concentration

 

 

V1

V2

V1

V2

V1

V2

V1

V2

(mg/l)

 

 

 

 

 

 

 

 

 

 

Control

4

4

7.5

7.5

7.6

7.6

7.8

7.8

7.9

7.9

6.25

4

4

7.8

7.8

7.9

7.9

7.9

8.0

8.0

8.0

12.5

4

4

-

-

-

-

-

-

-

-

25

4

4

-

-

-

-

-

-

-

-

50

4

4

-

-

-

-

-

-

-

-

100

4

4

-

-

-

-

-

-

-

-

 

Nominal

Number of

Oxygen content (mg/L) after

 

 

 

Concentrationfish

 

24h

 

48h

 

72h

 

96h

 

(mg/l)

 

 

V1

V2

V1

V2

V1

V2

V1

V2

Control

4

4

6.9

6.9

6.8

6.8

6.6

6.6

6.6

6.6

6.25

4

4

6.9

6.9

6.8

6.8

6.6

6.6

6.5

6.5

12.5

4

4

-

-

-

-

-

-

-

-

25

4

4

-

-

-

-

-

-

-

-

50

4

4

-

-

-

-

-

-

-

-

100

4

4

-

-

-

-

-

-

-

-

Test temperature

Nominal

Number of

Temperature (°C) after

 

 

 

 

Concentrationfish

 

24h

 

48h

 

72h

 

96h

 

(mg/l)

 

 

V1

V2

V1

V2

V1

V2

V1

V2

Control

4

4

23.4

23.4

23.5

23.5

23.3

23.3

23.5

23.5

6.25

4

4

23.4

23.4

23.5

23.5

23.3

23.3

23.5

23.5

12.5

4

4

-

-

-

-

-

-

-

-

25

4

4

-

-

-

-

-

-

-

-

50

4

4

-

-

-

-

-

-

-

-

100

4

4

-

-

-

-

-

-

-

-

Validity criteria fulfilled:
yes
Conclusions:
Based on nominal concentrations, experimental median lethal Concentrations [LC-50 (96 h)] for test chemical on Zebra Fish Danio rerio was determine to be 6.25 mg/l.
Executive summary:

Study was conducted to access the effect of test chemical on the growth of fish Danio rerio. Test conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test). The test substance was partially soluble in water. The in house water solubility in DI water was 21.6523 mg/L.Therefore, the stock solution was prepared by dissolving 5 g of the test substance in 5 liters of potable water (passed through reverse osmosis system) with continuous 72 hrs stirring. After stirring the stock analytically detected and solubility in RO water found to be 122.575 mg/L. By using this value of solubility following test concentrations were prepared 6.25mg/L, 12.5mg/L, 25mg/L,50mg/L,100mg/L ,respectively and Zebra Fish Danio rerio were exposed to these concentration for 96 hours. Bowl aquaria containing 2 liters of potable water (passed through reverse osmosis system) were loaded with 8 fishes. A static procedure was used for the study and it was conducted in compliance with the OECD guideline 203. After 96 hours of exposure to test item to various nominal test concentrations, LC50 was determine to be 6.25 mg/l. Based on the LC50, it can be consider that the chemical was toxic and can be consider to be classified as aquatic chronic 2 as per the CLP classification criteria.

Description of key information

Study was conducted to access the effect of test chemical on the growth of fish Danio rerio. Test conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test). The test substance was partially soluble in water. The in house water solubility in DI water was 21.6523 mg/L. Therefore, the stock solution was prepared by dissolving 5 g of the test substance in 5 liters of potable water (passed through reverse osmosis system) with continuous 72 hrs stirring. After stirring the stock analytically detected and solubility in RO water found to be 122.575 mg/L. By using this value of solubility following test concentrations were prepared 6.25mg/L, 12.5mg/L, 25mg/L,50mg/L,100mg/L ,respectively and Zebra Fish Danio rerio were exposed to these concentration for 96 hours. Bowl aquaria containing 2 liters of potable water (passed through reverse osmosis system) were loaded with 8 fishes. A static procedure was used for the study and it was conducted in compliance with the OECD guideline 203. After 96 hours of exposure to test item to various nominal test concentrations, LC50 was determine to be 6.25 mg/l. Based on the LC50, it can be consider that the chemical was toxic and can be consider to be classified as aquatic chronic 2 as per the CLP classification criteria.

Key value for chemical safety assessment

LC50 for freshwater fish:
6.25 mg/L

Additional information

Based on the short term toxicity studies available for the test chemical, studies were reviewed to determine the toxic nature of test chemical on the growth and mortality of fish. The studies are as mentioned below:

 

Study was conducted to access the effect of test chemical on the growth of fish Danio rerio. Test conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test). The test substance was partially soluble in water. The in house water solubility in DI water was 21.6523 mg/L. Therefore, the stock solution was prepared by dissolving 5 g of the test substance in 5 liters of potable water (passed through reverse osmosis system) with continuous 72 hrs stirring. After stirring the stock analytically detected and solubility in RO water found to be 122.575 mg/L. By using this value of solubility following test concentrations were prepared 6.25mg/L, 12.5mg/L, 25mg/L,50mg/L,100mg/L ,respectively and Zebra Fish Danio rerio were exposed to these concentration for 96 hours. Bowl aquaria containing 2 liters of potable water (passed through reverse osmosis system) were loaded with 8 fishes. A static procedure was used for the study and it was conducted in compliance with the OECD guideline 203. After 96 hours of exposure to test item to various nominal test concentrations, LC50 was determine to be 6.25 mg/l. Based on the LC50, it can be consider that the chemical was toxic and can be consider to be classified as aquatic chronic 2 as per the CLP classification criteria.

 

First study was supported by the second study. Study was conducted to access the effect of test chemical on the growth of fish Danio rerio. Test conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test). The test substance was sparingly soluble in water. Therefore, the stock solution was prepared by dissolving 2 gm of the test substance in 2 liter of potable water (passed through reverse osmosis system) with continuous stirring of 72 hrs. After the completion of 72 hrs stirring, the stock sample was analytically detected to calculate the actual solubility and the solubility obtained was then used for achieving test concentrations of 0.625 mg/L,1.25 mg/L,2.5 mg/L,5 mg/L & 10 mg/L, respectively. Bowl aquaria containing 2 liters of potable water (passed through reverse osmosis system) were loaded with 8 fishes. A static procedure was used for the study and it was conducted in compliance with the OECD guideline 203. The nominal concentration selected for the experiment was 0.625mg/L, 1.25mg/L, 2.5mg/L, 5mg/L, 10mg/L and Zebra fish were exposed to this concentration for 96 hours. The lethal concentrations LC50 was observed to be in the range of > 2.5 mg/l to < 5 mg/l. Based on nominal concentrations, experimental median lethal Concentrations [LC-50 (96 h)] for test chemical on Zebra Fish Danio rerio was determine to be > 2.5 mg/l to < 5 mg/l. Based on the LC50, it can be consider that the chemical was toxic and can be consider to be classified as aq aquatic chronic 2 as per the CLP classification criteria.

 

Thus based on the both above studies, chemical consider to be toxic and classified as aquatic chronic 2 as per the CLP classification criteria.