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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 result using OECD recommended guidelines.
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Principles of method if other than guideline:
This study was designed to assess the toxic effects of the test compound on the Zebra fish(Denio rerio) .Bowl aquaria containing 2 liters of potable water (passed through reverse osmosis system) were loaded with 8 fishes each. A static procedure was used for the study and it was conducted in compliance with the OECD guideline 203.
GLP compliance:
no
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Batch No.of test material: 0002
- Expiration date of the lot/batch: 2021-07-13
- Purity: 98.04%


STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Ambient Temp (23 to 27 °C) (Keep container tightly closed in a dry & well-ventilated place.Store in cool pla ce. Do not expose to air, light & moisture.)
- Stability under test conditions: Stable in recommended storage conditions
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 400 mg of the test substance in 4 liters of potable water (passed through reverse osmosis system) with 1 hr.continuous stirring for achieving test concentrations of 100 mg/L, respectively.

-Test water: Potable water (passed through reverse osmosis system) was used.
Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: Zebra Fish
- Source: Koyal aquarium,Nagpur.
- Length at study initiation (length definition, mean, range and SD): average 1.94 cm
- Weight at study initiation (mean and range, SD): 0.1494g (average)
- 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

ACCLIMATION
- Acclimation period: seven days
- Acclimation conditions: Photoperiod: 16 hours light and 8 hours dark, Temperature range: 23.6 °C, pH range: 7.35, DO range: 7.8 mg/l
- Feeding frequency during acclimation: Daily once
- Health during acclimation (any mortality observed): During last 48 hours = 0% mortality observed.

FEEDING DURING TEST
- Food type: Standard brand feed
- Frequency: Once, daily
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h
Test temperature:
24.3 °C
pH:
7.12
Dissolved oxygen:
7.8 mg/l
Nominal and measured concentrations:
100 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: The test vessels used were all plastic aquaria having the water capacity of 5 liters.
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: 2 liters.
- Aeration: Continuous
- Aeration in test vessels was provided 1 day before the start of experiment
- No. of organisms per vessel: 8 fishes


TEST MEDIUM / WATER PARAMETERS
- Hardness (mg of CaCO3): 148.5

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

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : LC50 was determined. Observations (mortality, visible symptoms, pH, Temperature, dissolved oxygen content) were recorded after 24, 48, 72 and 96 hours of the start of the experiment.

TEST CONCENTRATIONS
- Test concentrations: The test concentrations selected for the study were 100 mg/L.
Reference substance (positive control):
no
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 100 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:
LC0
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: No effects were observed on fish

Table 1: Mortality and symptoms

Nominal

Concentration

(mg/l)

Number of

fish

Mortality (cumulated) after

3 h

6 h

24h

48h

72h

96h

V1

V2

V1

V2

V1

V2

V1

V2

V1

V2

V1

V2

Control

4

4

-

-

-

-

-

-

-

-

-

-

-

-

100

4

4

-

-

-

-

-

-

-

-

-

-

-

-

 

 

Table 2: Visible Symptoms:

Nominal

Concentration

(mg/l)

Number of

fish

symptom (cumulated) after

3 h

6 h

24h

48h

72h

96h

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

100

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

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

 

Table 3: pH and oxygen content:

Nominal

Concentration

(mg/l)

Number of

fish

pH readings after

24h

48h

72h

96h

V1

V2

V1

V2

V1

V2

V1

V2

Control

4

4

8.01

8.02

8.12

8.15

8.23

8.25

8.31

8.35

100

4

4

8.21

8.22

8.31

8.32

8.37

8.39

8.41

8.42

 

Nominal

Concentration

(mg/l)

Number of

fish

pH readings after

24h

48h

72h

96h

V1

V2

V1

V2

V1

V2

V1

V2

Control

4

4

7.01

7.01

6.99

6.99

6.80

6.81

6.79

6.79

100

4

4

6.98

6.98

6.79

6.78

6.71

6.72

6.70

6.71

 

Table 4: Test temperature

Nominal

Concentration

(mg/l)

Number of

fish

pH readings after

24h

48h

72h

96h

V1

V2

V1

V2

V1

V2

V1

V2

Control

4

4

23.5

23.5

23.1

23.1

22.9

22.9

22.5

22.5

100

4

4

23.5

23.5

23.5

23.5

22.9

22.9

22.5

22.5

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 > 100 mg/L.

Executive summary:

The study was designed to assess the toxic effects of the test compound on the Zebra fish (Denio rerio). Bowl aquaria containing 2 liters of potable water (passed through reverse osmosis system) were loaded with 8 fishes each. A static procedure was used for the study and it was conducted in compliance with the OECD guideline 203 on Zebra fish (Danio rerio). The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 400 mg of the test substance in 4 liters of potable water (passed through reverse osmosis system) with 1 hr.continuous stirring for achieving test concentrations of 100 mg/L, respectively. The nominal concentration selected for the experiment were 100 mg/L and test fish were exposed to these concentration for 96 hours. The median lethal concentration (LC50) for test chemical on Danio rerio in a 96 hours study on the basis of mortality effect was determine to be >100 mg/L. Based on this LC50 value and according to CLP criteria for aquatic classification of the substance, it is concluded that the substance, does not exhibit short term toxicity to aquatic fish (Danio rerio).

LC0 (96 hours) (highest loading at which no mortality was observed) = 100 mg/L

LC50 (96 hours) Experimental = >100 mg/L

Description of key information

Study was designed to assess the toxic effects of the test compound on the Zebra fish (Denio rerio). Bowl aquaria containing 2 liters of potable water (passed through reverse osmosis system) were loaded with 8 fishes each. A static procedure was used for the study and it was conducted in compliance with the OECD guideline 203 on Zebra fish (Danio rerio). The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 400 mg of the test substance in 4 liters of potable water (passed through reverse osmosis system) with 1 hr.continuous stirring for achieving test concentrations of 100 mg/L, respectively. The nominal concentration selected for the experiment were 100 mg/L and test fish were exposed to these concentration for 96 hours. The median lethal concentration (LC50) for test chemical on Danio rerio in a 96 hours study on the basis of mortality effect was determine to be >100 mg/L. Based on this LC50 value and according to CLP criteria for aquatic classification of the substance, it is concluded that the substance, does not exhibit short term toxicity to aquatic fish (Danio rerio).

LC0 (96 hours) (highest loading at which no mortality was observed) = 100 mg/L

LC50 (96 hours) Experimental = >100 mg/L

Key value for chemical safety assessment

LC50 for freshwater fish:
100 mg/L

Additional information

Based on the effects observation on the mortality of fish by the chemical exposure, studies were reviewed from various experimental sources for test chemical were studied and mention as below:

In the first key study from experimental report, study was designed to assess the toxic effects of the test compound on the Zebra fish (Denio rerio). Bowl aquaria containing 2 liters of potable water (passed through reverse osmosis system) were loaded with 8 fishes each. A static procedure was used for the study and it was conducted in compliance with the OECD guideline 203 on Zebra fish (Danio rerio). The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 400 mg of the test substance in 4 liters of potable water (passed through reverse osmosis system) with 1 hr.continuous stirring for achieving test concentrations of 100 mg/L, respectively. The nominal concentration selected for the experiment were 100 mg/L and test fish were exposed to these concentration for 96 hours. The median lethal concentration (LC50) for test chemical on Danio rerio in a 96 hours study on the basis of mortality effect was determine to be >100 mg/L. Based on this LC50 value and according to CLP criteria for aquatic classification of the substance, it is concluded that the substance, does not exhibit short term toxicity to aquatic fish (Danio rerio).

LC0 (96 hours) (highest loading at which no mortality was observed) = 100 mg/L

LC50 (96 hours) Experimental = >100 mg/L

 

First study was supported by the second study from secondary source. A marine toxicity test was conducted to determine the acute effect of test chemical on sheepshead minnows (Cyprinodon variegatus) in a static system for 96 hours study. The test was conducted in 3.8 liter covered glass jars each of which contained a final volume of 3 L. There were no aeration. Based on the results of range finding test with mysid shrimp, fish were definitively tested at maximum concentration of 1000 mg/L. Therefore 1000 mg/l test concentration used in study. This test concentration was prepared by adding an appropriately weighed amount of test chemical directly to seawater in each test jar. Control jars contained only seawater. Test chemical was not toxic to sheepshead minnows in a test concentration of 1,000 mg/l. After 96 hours of exposure, there was no mortality in the 1,000 mg/l test concentration and also in the control. Fish in both the control and test concentration appeared to be in good condition throughout the test. Therefore, the lethal concentration (LC0) of test chemical on Cyprinodon variegatus was considered to be 1000 mg/L.

 

Thus on the basis of above effects on the mortality of fish, it was concluded that the chemical was nontoxic and not classified as per the CLP classification criteria.