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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:
12/12/1996 - 23/09/1998
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
- Concentrations: Nominal: 0.50, 0.75, 1.1, 1.7 and 2.5 mg/l
- Sampling method: Duplicate 10-mL samples of each new WAF solution were collected at 0, 24, 48 and 72 hours and analyzed for OTNE by LSC. Equal volumes of old test solutions were collected from both replicates of the control, vehicle blank, and each treatment level at 24, 48, 72 and 96 hours.
- Sample storage conditions before analysis: No data
Vehicle:
yes
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The dosing solution was an isotopic dilution containing 14C-CPD 98663 and OTNE. The solution was prepared by quantitatively transferring a 1.0 mCi sample of 14C-CPD 98663 with OTNE to a volume of 10.0 mL. This standard solution was identified as PR-01554 and was analyzed by liquid scintillation counting (LSC) analysis to have an active ingredient concentration of 9.60 x 10^5 µg/mL. A 0.520-mL aliquot of this solution was brought to a 10.0 mL total volume with acetone to prepare the finai dosing solution with a concentration of 4.99 x 10^4 µg/mL as analyzed by LSC. This solution was identified as R-01870 and was used to prepare all of the exposure solutions at each time period.

Test solutions were prepared individually for each treatment by adding an appropriate amount of the 4.99 x 10^4 µg/mL dosing solution to 9.0 L of dilution water in a 9.5-L glass bottle. An additional volume of acetone was added to levels 1-4 to produce acetone concentrations equal to that in level 5 (50 µL/L). Each bottle contained a Teflon-coated stir bar and the contents of the bottles were stirred for 20 ± 1 hours. Solutions were stirred so that a vortex of <25 % of the bottle depth was achieved. When stirring was terminated, the phases were allowed to separate for 1-1.5 hours. A portion of the water phase (water accommodated fraction - WAF) in each bottle was removed via a glass siphon tube and used to fill the respective test chambers. New WAF solutions were prepared and used to renew the test solutions every 24 hours during the test.
- Controls: yes controls and vehicle controls
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): 50 ul/l
- Evidence of undissolved material (e.g. precipitate, surface film, etc): No, when stirring was terminated, the phases were allowed to separate for 1-1.5 hours. A portion of the water phase (water accommodated fraction - WAF) in each bottle was removed via a glass siphon tube and used to fill the respective test chambers.
Test organisms (species):
Lepomis macrochirus
Details on test organisms:
TEST ORGANISM
- Common name: Bluegill
- Strain: No data
- Source: Northeastern aquatics, Rhinebeck, New York
- Age at study initiation (mean and range, SD): No data
- Length at study initiation (length definition, mean, range and SD): standard length 18 +/- 1.3 mm, total length 23 +/- 1.5 mm
- Weight at study initiation (mean and range, SD): blotted wet weight 0.130 +/- 0.034 g
- Method of breeding: The bluegill were held in dilution water under environmental conditions essentially the same as those in the definitive test, until they were used for the test. Fish were fed brine shrimp nauplii twice daily during the holding period. No diseases were observed or treated during the 20-day period prior to definitive testing. Two days prior to test initiation, approximately 200 individuals were placed into acclimation. The bluegill were not fed during the acclimation period. Acclimation water temperature ranged from 24 to 25 ° C. During the 48-hour acclimation period no mortality was observed and all fish appeared to be in good condition.
- Feeding during test: no feeding during test


ACCLIMATION
- Acclimation period: 48 hours
- Acclimation conditions (same as test or not): same as test
- Type and amount of food: No feeding during acclimation
- Health during acclimation (any mortality observed): fish appeared to be in good condition
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96
Hardness:
136 - 142 mg/l CaCO3
Test temperature:
New parent solutions and replicate old solutiona: 22.6 - 23.6 degrees Celsius; replicate new solutions: 23.5 - 24.5 degrees Celsius.
pH:
8.0 - 8.3
Dissolved oxygen:
New parent solutions: 7.7 - 7.9 mg/l (94 - 96% of saturation); replicate old solutions: 5.5 - 7.7 mg/l (67 - 94% saturation).
Salinity:
Not relevant
Nominal and measured concentrations:
Nominal: 0.50, 0.75, 1.1, 1.7 and 2.5 mg/l
Measured (0h): 0.566, 0.840, 1.26, 1.87 and 2.72 mg/l; Measured (24h, old): 0.484, 0.728, 1.05, 1.63 and 2.71 mg/l
Measured (24h): 0.550, 0.836, 1.21, 1.89 mg/l; Measured (48h, old): 0.520, 0.763, 1.16 and 1.84 mg/l
Measured (48h): 0.550, 0.838, 1.24, 1.87 mg/l; Measured (72h, old): 0.541, 0.827, 1.22 and 1.89 mg/l
Measured (72h): 0.544, 0.851, 1.27 mg/l; Measured (96h old): 0.559, 0.847 and 1.26 mg/l
Measured (mean): 0.54, 0.82, 1.2, 1.8 and 2.7 mg/l
Missing values are caused by termination of experiments after 100% mortality
Details on test conditions:
TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: glass, 3.8 liter filled with as little headspace as possible
- Aeration: No data
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2
- No. of vessels per vehicle control (replicates): 2
- Biomass loading rate: 0.17 g/l


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: blend of naturally hard well water with demineralized well water
- Total organic carbon: <1.00 mg/l
- Metals:
Al <0.044 mg/l
As 0.0057 mg/l
B 0.301 mg/l
Cd <0.00042 mg/l
Cr <0.0013 mg/l
Co <0.0058 mg/l
Co <0.00045 mg/l
Fe <0.015 mg/l
Pb <0.0034 mg/l
Hg <0.000023 mg/l
Ni <0.0078 mg/l
Se <0.0037 mg/l
Ag <0.00081 mg/l
Zn 0.021 mg/l
- Pesticides: all < 3.8 ug/l
- Alkalinity: 150 - 154 mg/l CaCO3
- Conductivity: 302 - 304 uS/cm
- Culture medium different from test medium: No
- Intervals of water quality measurement: 24 hours


OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16/8 light/dark
- Light intensity: No data


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality and sublethal responses were observed every 24 hours


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.5
- Range finding study
- Test concentrations: 0.10, 1.0, 2.0 and 10.0 mg/l
- Results used to determine the conditions for the definitive study: After 96 hours of exposure, mortality was 80% at 10 mg/L and 0% at test concentrations < 2 mg/L. Mortality in the control and vehicle blank was 0%. Based upon the results of this test and solubility of the test substance, a nominal concentration range of 0.50, 0.75, 1.1, 1.7 and 2.5 mg/L was selected for the definitive test.
Reference substance (positive control):
no
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
1.3 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
other: radioactive compound
Basis for effect:
mortality
Remarks on result:
other: 95% CL: 1.2 - 1.5 mg/l
Duration:
24 h
Dose descriptor:
LC50
Effect conc.:
2.2 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
other: radioactive compound
Basis for effect:
mortality
Remarks on result:
other: 95% CL: 1.8 - 2.7 mg/l
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
1.8 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
other: Radioactive compound
Basis for effect:
mortality
Remarks on result:
other: 95% CL: 1.6 - 2.0 mg/l
Duration:
72 h
Dose descriptor:
LC50
Effect conc.:
1.4 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
other: Radioactive compound
Basis for effect:
mortality
Remarks on result:
other: 95% CL: 1.3 - 1.5 mg/l
Details on results:
- Mortality of control: 0%
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: No
- Effect concentrations exceeding solubility of substance in test medium: No
Reported statistics and error estimates:
Statistical analysis of the concentration versus effect data (mortality) was performed by a computerized LC50 program using Trimmed Spearman-Karber techniques. The slope of the concentration-response line was calculated by transforming percent mortality to probit values and plotting probit values against the log of the concentration. Linear regression was used for the calculation of the slope of the concentration-response line.

The slope of the dose-response line was 13.67.
Validity criteria fulfilled:
yes
Remarks:
mortality in controls <10%, oxygen saturation > 60% during test, test substance concentration maintained within 20% of nominal concentration, constant testing conditions maintained
Conclusions:
The 96h-LC50 of OTNE towards Lepomis macrochirus is 1.3 mg/l.
Executive summary:

The toxicity of OTNE towards Lepomis macrochirus was investigated according to methodology comparable to OECD Guideline 203 under GLP. Fish were exposed to water accommodated fractions (WAF) of 14C-labelled OTNE with nominal concentrations of 0.50, 0.75, 1.1, 1.7 and 2.5 mg/l in a semi-static system and observed for 96 hours. Validity criteria for the test were met. The 96h-LC50 was found to be 1.3 mg/l (95% CL 1.2 - 1.5 mg/l) based on mean measured concentrations.

Description of key information

The toxicity of OTNE towards Lepomis macrochirus was investigated according to methodology comparable to OECD Guideline 203 under GLP. Fish were exposed to water accommodated fractions (WAF) of 14C-labelled OTNE with nominal concentrations of 0.50, 0.75, 1.1, 1.7 and 2.5 mg/l in a semi-static system and observed for 96 hours. Validity criteria for the test were met. The 96h-LC50 was found to be 1.3 mg/l (95% CL 1.2 - 1.5 mg/l) based on mean measured concentrations.
    
    
    
    

The toxicity of OTNE towards Lepomis macrochirus was investigated according to methodology comparable to OECD Guideline 203 under GLP. Fish were exposed to water accommodated fractions (WAF) of 14C-labelled OTNE with nominal concentrations of 0.50, 0.75, 1.1, 1.7 and 2.5 mg/l in a semi-static system and observed for 96 hours. Validity criteria for the test were met. The 96h-LC50 was found to be 1.3 mg/l (95% CL 1.2 - 1.5 mg/l) based on mean measured concentrations.

The toxicity of OTNE towards Lepomis macrochirus was investigated according to methodology comparable to OECD Guideline 203 under GLP. Fish were exposed to water accommodated fractions (WAF) of 14C-labelled OTNE with nominal concentrations of 0.50, 0.75, 1.1, 1.7 and 2.5 mg/l in a semi-static system and observed for 96 hours. Validity criteria for the test were met. The 96h-LC50 was found to be 1.3 mg/l (95% CL 1.2 - 1.5 mg/l) based on mean measured concentrations.

The toxicity of OTNE towards Lepomis macrochirus was investigated according to methodology comparable to OECD Guideline 203 under GLP. Fish were exposed to water accommodated fractions (WAF) of 14C-labelled OTNE with nominal concentrations of 0.50, 0.75, 1.1, 1.7 and 2.5 mg/l in a semi-static system and observed for 96 hours. Validity criteria for the test were met. The 96h-LC50 was found to be 1.3 mg/l (95% CL 1.2 - 1.5 mg/l) based on mean measured concentrations.

Key value for chemical safety assessment

LC50 for freshwater fish:
1.3 mg/L

Additional information