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Ecotoxicological information

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:
October 1987
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
GLP and test guideline compliant study, oxygen concentration too low at higher concentrations, good documentation, no dose verification performed
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
GLP compliance:
yes
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The definitive test concentrations were obtained by transferring appropriate weights of test item directly to the test chambers.
- Controls: blank controls
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): NA
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): NA
- Evidence of undissolved material (e.g. precipitate, surface film, etc): There was some undissolved compound in each test vessel at 0-hour. All material had completely dissolved after 24 hours, although the 1000 mg/L solution was slightly cloudy.
- Test solution preparation:
The compound was allowed to stay in the water sample until it became partly dissolved at which time the partly dissolved compound solution was added to the test vessels and vigorously stirred. Fish were not added to the test solutions until the very end of the 30 minutes time limit in order to allow for more complete dissolution of the test compound. All test concentrations were based on teh total compound, i.e., not corrected for purity or undissolved material.
Test organisms (species):
Lepomis macrochirus
Details on test organisms:
TEST ORGANISM
- Common name: bluegill sunfish
- Source: Osage Catfisheries, Osage Beach
- Length at study initiation (length definition, mean, range and SD): 22 (+/- 1.1 ) mm
- Weight at study initiation (mean and range, SD): 0.23 (+/- 0.043) g/fish
- Feeding during test: the fish were not fed during the exposure



ACCLIMATION
- Acclimation period: at least 14 days
- Acclimation conditions (same as test or not): same as test
- Type and amount of food: standard commercial fish food
- Feeding frequency: daily, discontinued 48 to 96 hours prior to testing


QUARANTINE (wild caught)
- Duration: at least 14 days
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Hardness:
40-48 mg/L as CaCO3
Test temperature:
21 - 22°C
pH:
6.7 - 7.3
Dissolved oxygen:
1.1 - 8.5 mg/L
Nominal and measured concentrations:
nominal concentrations: 0 (control), 240, 320, 420, 560, 750 and 1000 mg CT-316-87/L
Details on test conditions:
TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 5 gallon glass vessels containing 15 L test solution
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): NA
- Biomass loading rate: 0.15 g/L


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: soft reconstituted water
Following amounts were added to deionized water:
NaHCO3: 48 mg/L
CaSO4*2H2O: 30 mg/L
MgSO4: 30 mg/L
KCl: 2 mg/L
- Metals: measured, levels below levels of concern
- Pesticides: measured, levels below levels of concern
- Alkalinity: 25 - 35 mg/L as CaCO3
- Intervals of water quality measurement: measured at 24, 48 and 96 hours


OTHER TEST CONDITIONS
- Adjustment of pH: no



EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
mortality and behavioural changes

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.3
- Justification for using less concentrations than requested by guideline: NA
- Range finding study: two range finding studies were performed:
- Test concentrations:
first range finder: 1, 10, 100 mg CT-316-87/L
second range finder: 1000 mg CT-316-87/L
third range finder: 320 mg CT-316-87/L
- Results used to determine the conditions for the definitive study:
first range finder: 1, 10, 100 mg CT-316-87/L with 0/5, 0/5, 0/5 dead fish after 96 hours;
second range finder: 1000 mg CT-316-87/L with 3/5 dead fish after 48 hours
third range finder: 320 mg CT-316-87/L with 0/5 dead fish after 96 hours
Reference substance (positive control):
no
Duration:
24 h
Dose descriptor:
LC50
Effect conc.:
777.6 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
mortality (fish)
Duration:
24 h
Dose descriptor:
LC50
Effect conc.:
810 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: 95% C.I.: 710 - 960 mg test material/L
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
451.2 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
mortality (fish)
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
470 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: 95% C.I.: 420 - 530 mg test material/L
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
451.2 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
mortality (fish)
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
470 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: 95% C.I.: 420 - 530 mg test material/L
Details on results:
- Behavioural abnormalities: none reported
- Observations on body length and weight: NA
- Other biological observations: no behavioural changes at 240 mg CT-316-87/L
- Mortality of control: none
- Other adverse effects control: none
- Abnormal responses: At concentrations of 320 mg/L and higher surfacing, loss of equilibrium, fish on the bottom of the test vessel, quiescence and / or laboured respiration was observed.
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values:
After preparation of the test solutions there was undissolved compound in all (treated) test vessels. After 24 hours, this compound was dissolved in all test vessels. At 1000 mg/L the test solution was slightly cloudy. After 48 hours and for the reminder of the study, all test solutions from 240 to 1000 mg/L were slightly cloudy. The opacity increased slightly with the increase in concentration.
- Effect concentrations exceeding solubility of substance in test medium: Apparently, all test solutions were above the limit of water solubility.
Reported statistics and error estimates:
The LC50 values for CT-316-87 were determined using the probit method.
Sublethal observations / clinical signs:

Table 1 Nominal concentration of CT-316-87 and corresponding concentrations of CH-SSS in the test solutions (calculated for 96% purity)

 

Nominal concentration

[mg CT-316-87/L]

Nominal concentration

[mg a.i./L]

0 (Control)

0 (Control)

240

230.4

320

307.2

420

403.2

560

537.6

750

720

 1000  960

 

 

Table 2   Temperature, dissolved oxygen and pH values measured in the test vessels at different time intervals

 

Nominal concentration

[mg CT-316-87 /L]

0 hours

48 hours

96 hours

Temp

[°C]

DO

[mg/L]

pH

Temp

[°C]

DO

[mg/L]

pH

Temp

[°C]

DO

[mg/L]

pH

0 (Control)

21

8.5

7.5

22

6.5

7.3

22

6.2

7.2

240

21

8.5

7.7

22

3.2

6.9

22

4.0

6.9

320

-

-

-

22

2.1

6.8

22

3.4

6.8

 420  21  8.5  7.7  22  1.2  6.7  22  3.3  6.8
 560  -  -  -  22  1.1  6.7  22  1.6  6.7
 750  -  -  -  -
 1000  21  8.5  7.6  -

 

 

Table 3 Percent mortality at the individual observation intervals

 

Nominal concentration

[mg CT-316-87 /L]

Cumulative Percent Mortality

24 hours

48 hours

96 hours

0 (Control)

0

0

0

240

0

0

0

320

0

0

0

420

0

20

20

560

10

90

100

750

20

100

100

 1000  90  100  100

 

Validity criteria fulfilled:
yes
Remarks:
the control validity criteria were fulfilled, the DO concentration criteria not at higher concentrations
Conclusions:
96-hour LC50 (Lepomis macrochirus)=451.2 mga.i./L
Executive summary:

The acute toxicity of CT-316 -87 - which contains 96% a.i. - to bluegill sunfish, Lepomis macrochirus, was determined in a static test. The test was performed according to OECD 203. Deviations to the guideline were significant oxygen reduction in the higher test groups which might have resulted in mortality. The test solutions were not aerated. No dose-verification analysis was performed. The test was performed with a control and nominal 240, 320, 420, 560, 750 and 1000 mg CT-316 -87/L corresponding to 230.4, 307.2, 403,2, 537,6, 720 and 960 mg a.i./L. During the study, the temperature of the test solutions ranged from 21 to 22°C. During the study the oxygen concentration declined with increasing test substance concentration. The lowest dissolved oxygen concentration measured was 1.1 mg/L. After 96 hours of exposure, 0, 0, 0, 2, 9, 10 and 10 out of 10 fish had died in the control, and 240, 320, 420, 560, 750 and 1000 mg CT-316 -87/L treatment groups. The 96 h LC50 was determined to be 470 mg CT-316 -87/L corresponding to 451.2 mg a.i./L. The concentrations were based on nominal concentrations.

The results of this study need to be interpreted with care since the dissolved oxygen concentrations might have had a significant impact on the mortality observed during the test. However, since the potential mortality caused by oxygen reduction is considered to increase the mortality and hence represents the worst case scenario the results are considered to be reliable with restrictions for the risk assessment.

Description of key information

LC50 bluegill sunfish: 451.2 mga.i./L

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Dose descriptor:
LC50
Effect concentration:
451.2 mg/L

Additional information

The study from Cytec (Bowman 1987) was performed under GLP was scored as Klimisch 2 study. The acute toxicity of CT-316 -87- which contains 96% a.i. - to bluegill sunfish, Lepomis macrochirus, was determined in a static test. The test was performed according to OECD 203. Deviations to the guideline were significant oxygen reduction in the higher test groups which might have resulted in mortality. The test solutions were not aerated. No dose-verification analysis was performed. The test was performed with a control and nominal 240, 320, 420, 560, 750 and 1000 mg CT-316 -87/L corresponding to 230.4, 307.2, 403,2, 537,6, 720 and 960 mg a.i./L. During the study, the temperature of the test solutions ranged from 21 to 22°C. During the study the oxygen concentration declined with increasing test substance concentration. The lowest dissolved oxygen concentration measured was 1.1 mg/L. After 96 hours of exposure, 0, 0, 0, 2, 9, 10 and 10 out of 10 fish had died in the control, and 240, 320, 420, 560, 750 and 1000 mg CT-316 -87/L treatment groups. The 96 h LC50 was determined to be 470 mg CT-316 -87/L corresponding to 451.2 mg a.i./L.The concentrations were based on nominal concentrations.

The results of this study need to be interpreted with care since the dissolved oxygen concentrations might have had a significant impact on the mortality observed during the test. However, since the potential mortality caused by oxygen reduction is considered to increase the mortality and hence represents the worst case scenario the results are considered to be reliable with restrictions for the risk assessment.