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Short-term toxicity to fish

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Endpoint:
short-term toxicity to fish
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
Mar 12, 2001 to Mar 16, 2001
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
The reliability rating is a 1 because the study followed a standard guideline, which describes a procedure designed to evaluate this endpoint, and the results were reviewed for reliability and assessed as valid. The study was also conducted under GLP.
Justification for type of information:
The justification for read across is provided as an attachment in IUCLID Section 13.
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
according to guideline
Guideline:
EPA OPP 72-1 (Fish Acute Toxicity Test)
Deviations:
yes
Remarks:
Minor deviations that were not believed to have affected the outcome of the study.
Principles of method if other than guideline:
Due to the complex nature and limited water solubility of the test substance, the following exceptions to the guideline apply for this study: The concentration of the test substance in solution was not determined prior to use. It was deemed more appropriate to prepare individual treatment solutions by adding the test substance to dilution water and removing the WAF of each mixture for testing than to prepare dilutions of a stock solution. The initial concentration of the test substance was not maintained at 80% in the 2.0 mg/L (78% maintained) and 10 mg/L (76% maintained) treatments throughout the test. Fish were smaller (average 3.3 cm) than the guideline minimum recommended size of 4 cm. Smaller fish were used to help maintain the dissolved oxygen, due to the study design (no headspace). The pH varied by more than one unit during the test, range was 6.3 to 7.8.
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
Samples were removed from the fresh solutions at initiation, on Day 0 and 3. Samples were also removed from the old solutions on Day 1, and at termination on Day 4.
Vehicle:
no
Details on test solutions:
Individual treatment solutions were prepared as Water Accommodated Fractions (WAFs). This included adding test substance to 13 L glass aspirator bottles, which contained 13.5 L of laboratory dilution water and mixing for 24 hours with a motorized stirrer at a rate that minimized the vortex so that no emulsion was formed. After mixing, the WAFs were allowed to settle. The test solutions were then siphoned from the bottom of the mixing vessel and used to prepare the individual treatment replicates.
Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
Fish supplied by Pierce Associates, Inc., West Buxton, ME; age = 10 weeks old; mean wt. = 0.219 g; mean total length = 3.3 cm; test loading = 0.243 g of fish/L.
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Post exposure observation period:
None
Hardness:
Water hardness was 104 mg/L as CaCO3.
Test temperature:
Mean Test temperature was 13.9 Deg C.
pH:
The pH ranged from 6.3 to 7.8 during the study.
Dissolved oxygen:
Dissolved oxygen = 6.2 to 9.1 mg/L
Salinity:
Freshwater
Nominal and measured concentrations:
Nominal treatment levels were: control, 0.43, 0.98, 2.0, 4.8, and 10 mg/L, which measured 0, 0.12, 0.28, 0.51, 0.71, and 0.61 mg/L, respectively, as calculated using the mean of new solution values.
Details on test conditions:
Lighting was 727 to 736 Lux with 16 hrs light and 8 hrs dark.

Two replicates of each treatment were tested in 4 L glass aspirator bottles with 5 fish in each tank. New WAFs for each treatment were prepared as described above for each daily renewal. Renewal of test solution was performed by siphoning out approximately 80% of the old solution and refilling with freshly prepared WAF.
Reference substance (positive control):
no
Duration:
24 h
Dose descriptor:
LL50
Effect conc.:
5.9 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
48 h
Dose descriptor:
LL50
Effect conc.:
5.7 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
72 h
Dose descriptor:
LL50
Effect conc.:
4.3 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Key result
Duration:
96 h
Dose descriptor:
LL50
Effect conc.:
3.6 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Details on results:
No control mortalities or abnormal observations were recorded during the study.
Reported statistics and error estimates:
The 24 to 96-hr LL50 value were determined using a maximum likelihood analysis based on D.J. Finney (1971).
Sublethal observations / clinical signs:

96-hour LL50 = 3.6 mg/L (95% CI Could Not Calculate) based upon nominal values
96-hour LC50 = 0.46 mg/L (95% CI 0.37 to 0.53) based upon mean measured values

Nominal  Measured Fish Total Mortality
Conc. (mg/L) Conc. (mg/L) (@ 24, 48, 72, 96 hrs)**
     
Control Control 0, 0, 0, 0
0.43 0.12 0, 0, 0, 0
0.98 0.28 0, 0, 0, 1
2.00 0.51 6, 7, 7, 7
4.80 0.71 9, 9, 10, 10
10.00 0.61 5, 5, 7, 8

*Concentrations based upon mean measured values of the test solutions
**10 fish added to each of the control and treatment levels at test initiation

Validity criteria fulfilled:
yes
Conclusions:
Water accommodated fractions of hydrocarbons, C10-C13, aromatics, <1% naphthalene, produced a 96-hour LL50 value of 3.6 mg/l with rainbow trout (Oncorhynchus mykiss).
Executive summary:

Water accommodated fractions of hydrocarbons, C10-C13, aromatics, <1% naphthalene, produced a 96-hour LL50 value of 3.6 mg/l with rainbow trout (Oncorhynchus mykiss).

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1993
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
The study was given a reliability rating of 1 because it followed a standard test guideline, which describes a procedure to evaluate this endpoint, and the results were reviewed for reliability and assessed as valid. The study was also conducted under GLP.
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
However, analysis of the test media was restricted to the media produced at higher loading rates. All media produced at loading rates less 2mg/L were not analysed because they were below the limit of determination (2.5 mg/L) of the analytical method.
Vehicle:
no
Details on test solutions:
Individual treatment solutions were prepared as Water Accommodated Fractions (WAFs). This included adding test substance to glass vessels, which contained laboratory dilution water and mixing for approximately 24 hours with a magnetic stirrer to produce a series of geometrically-spaced loading rates (mg/L). After mixing, the WAFs were allowed to settle for an hour. The aqueous phase was then drawn off and transferred to 11 litre full-volume, spherical glass vessels which were completely filled. An additional vessel received dechlorinated water only and served as a control.
Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
O.mykiss fingerlings were supplied by Exmoor Trout Farm, North Molton, Devon, UK; mean wt. = 0.96 g; mean total length = 4.4 cm.
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Hardness:
Water hardness during the test ranged from 280 - 314 mg/L as CaCO3.
Test temperature:
Test temperature was 15 – 16 ºC with a 16 h : 8 h light : dark lighting regime.
pH:
The pH ranged from 6.8 to 8.5 during the test.
Dissolved oxygen:
Dissolved oxygen = 9.4 to 10.5 mg/L for "new" solutions and 7.5 to 9.3 mg/L for "old" solutions.
Salinity:
freshwater
Nominal and measured concentrations:
Nominal treatment levels were: control, 0.2, 0.7, 2.0, 5.0, 17 and 50 mg/L. Analysis confirmed concentrations at t=0 of 4.3, 7.5 and 9.5 mg/L for the nominal loading rates of 5.0, 17 and 50 mg/L, respectively. As there were fish mortalities at these loading rates, no further samples were available for analysis. All media produced at loading rates less than 2mg/L were not analysed because they were below the limit of determination (2.5 mg/L)
of the analytical method.
Details on test conditions:
7 fish were placed in each test vessel, which was then sealed with a glass plate, ensuring that all air was excluded. New WAFs for each treatment were prepared as described above for each daily renewal. Surviving fish were transferred to the vessel containing freshly prepared WAF. The fish were not fed during the test.

Test temperature was 15 – 16 ºC with a 16 h : 8 h light : dark lighting regime. Dissolved oxygen = 9.4 to 10.5 mg/L for "new" solution and 7.5 to 9.3 mg/L for "old" solution. The pH ranged from 6.8 to 8.5 during the test. Water hardness during the test ranged from 280 - 314 mg/L as CaCO3. The water used was harder than the preferred range (10 - 250 mg/L) . However, the hardness measured is normal for this laboratory and does not affect fish survival.
Reference substance (positive control):
no
Key result
Duration:
96 h
Dose descriptor:
LL50
Effect conc.:
>= 2 - <= 5 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Details on results:
See Table Below
Reported statistics and error estimates:
Statistical Method: None required

Loading rates causing a 50% effect were determined by visual inspection of the data. They are expressed as being within a range bounded by
loading rates that were tested. This is considered more appropriate for test materials that are mixtures of substances of different solubilities
than is an interpolative statistical technique
Sublethal observations / clinical signs:

Nominal Loading Measured Fish Total Mortality
Conc. (mg/l) Conc. (mg/l) (@ 24, 48, 72, 96 hrs)
     
Control Control 0, 0, 0, 0
0.2 < LoD 0, 0, 0, 0
0.7 < LoD 1, 1, 1, 1
2.0 < LoD 0, 0, 0, 0
5.0 4.3 0, 7, 7, 7
17.0 7.5 7, 7, 7, 7
50.0 9.5 7, 7, 7, 7
Validity criteria fulfilled:
yes
Conclusions:
Water accommodated fractions of hydrocarbons, C10, aromatics, >1% naphthalene, produced a 96-hour LL50 value between 2 and 5 mg/L with rainbow trout, Oncorhynchus mykiss.
Executive summary:

Water accommodated fractions of hydrocarbons, C10, aromatics, >1% naphthalene, produced a 96-hour LL50 value between 2 and 5 mg/L with rainbow trout, Oncorhynchus mykiss.

Endpoint:
short-term toxicity to fish
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
13-Jan-1992 to 17-Jan-1992
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
The reliability rating is a 1 because the study followed a standard guideline, which describes a procedure designed to evaluate this endpoint, and the results were reviewed for reliability and assessed as valid. The study was also conducted under GLP.
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
according to guideline
Guideline:
EPA OPP 72-1 (Fish Acute Toxicity Test)
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
Samples were removed from the fresh solutions at initiation, on Day 1, 2, and 3. Samples were also removed from the old solutions on Day 1, and at termination on Day 4.
Vehicle:
no
Details on test solutions:
Individual treatment solutions were prepared as Water Accommodated Fractions (WAFs). This included adding test substance to 20 L glass carboys, which contained 20 L of laboratory dilution water and mixing for 24 hours with a motorized stirrer at a rate that minimized the vortex so that no emulsion was formed. After mixing, the WAFs were allowed to settle. The test solutions were then siphoned from the bottom of the mixing vessel and used to prepare the individual treatment replicates.
Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
Fish supplied by Aquatic Research Organisms, Hampton, NH; age = 11 weeks old; mean wt. = 0.575 g; mean total length = 3.4 cm; test loading = 0.553 g of fish/L.
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Post exposure observation period:
None
Hardness:
Water hardness was 44 mg/L as CaCO3.
Test temperature:
Test temperature was 12.0 +/- 1.0 Deg C.
pH:
The pH ranged from 7.6 to 7.7 during the study.
Dissolved oxygen:
Dissolved oxygen = 8.9 to 10.0 mg/L for "new" solution and 7.0 to 8.7 mg/L for "old" solution.
Salinity:
Freshwater
Nominal and measured concentrations:
Nominal treatment levels were: control, 0.5, 1.0, 5.0, 10.0, and 50 mg/L, which measured <0.076, 0.481, 0.872, 3.82, 6.11, and 18.2 mg/L, respectively, as calculated using the mean of new solution values.
Details on test conditions:
Lighting was 68 to 70 ft candles with 16 hrs light and 8 hrs dark.

Four replicates of each treatment were tested in 5.2 L glass tanks with 5 fish in each tank. New WAFs for each treatment were prepared as described
above for each daily renewal. Renewal of test solution was performed by siphoning out approximately 86% of the old solution and refilling with
freshly prepared WAF.
Reference substance (positive control):
no
Key result
Duration:
96 h
Dose descriptor:
LL50
Effect conc.:
3 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: 95% CI = 1.0 - 5.0
Details on results:
No control mortalities or abnormal observations were recorded during the study.
Reported statistics and error estimates:
The 24-hr LC50 value was determined using the Probit procedure of SAS. The 48-hr LC50 value was determined using the Spearman-Karber Estimation Method. The 72 and 96-hr LC50 values were determined using the Binomial Method.
Sublethal observations / clinical signs:

96-hour LL50 = 3.0 mg/L (95% CI 1.0 to 5.0) based upon nominal values
96-hour LC50 = 2.34 mg/L (95% CI 0.87 to 3.82) based upon measured values of new solutions

Nominal  Measured Fish Total Mortality
Conc. (mg/L) Conc. (mg/L) (@ 24, 48, 72, 96 hrs)**
     
Control Control 0, 0, 0, 0
0.5 0.48 0, 0, 0, 0
1.0 0.87 0, 0, 0, 0
5.0 3.82 7, 10, 20, 20
10.0 6.11 19, 20, 20, 20
50.0 18.2 20, 20, 20, 20

*Concentrations based upon measured values of new solutions
**20 fish added to each of the control and treatment levels at test initiation

Validity criteria fulfilled:
yes
Conclusions:
Water accommodated fractions of hydrocarbons, C10-C13, aromatics, >1% naphthalene, produced a 96-hour LL50 of 3.0 mg/l with rainbow trout (Oncorhynchus mykiss).
Executive summary:

Water accommodated fractions of hydrocarbons, C10-C13, aromatics, >1% naphthalene, produced a 96-hour LL50 of 3.0 mg/l with rainbow trout (Oncorhynchus mykiss).

Description of key information

There is data available for this substance. Additionally, key and supporting data is available for the following structural analogues: Hydrocarbons, C10-C13, aromatics, <1% naphthalene and Hydrocarbons, C10-C13, aromatics, >1% naphthalene. The data for these substances is presented in the dossier. The data is read across to these substances based on analogue read across and a discussion and report on the read across strategy is provided as an attachment in IUCLID Section 13.

Hydrocarbons, C10-C13, aromatics, <1% naphthalene, presented a 96-h LL50 (mortality) for Oncorhynchus mykiss of 3.6 mg/L.

Hydrocarbons, C10, aromatics, >1% naphthalene presented a 96-h LL50 (mortality) for Oncorhynchus mykiss of between 2 and 5 mg/L

Hydrocarbons, C10-C13, aromatics, >1% naphthalene, presented a 96-h LL50 (mortality) for Oncorhynchus mykiss of 3.0 mg/L.

Hydrocarbons, C10, aromatics, >1% naphthalene, presented a 96-h LL50 (mortality) for Oncorhynchus mykiss of 14 mg/L.

Key value for chemical safety assessment

Additional information

Four study reports were available and input as endpoint records. The key studies are summarised below.

The study from Exxon (2006) examined the short-term toxicity of the test substance Hydrocarbons, C10-C13, aromatics, <1% naphthalene, to Oncorhynchus mykiss. Groups of 10 fish were added to each of the control and treatment levels at test initiation, with nominal treatment levels of control, 0.43, 0.98, 2.0, 4.8, and 10 mg/L. The fish were exposed to the test substance for 96h, and fish were observed for mortality at 24, 48, 72, 96 hrs. Water accommodated fractions of hydrocarbons, C10-C13, aromatics, <1% naphthalene, produced a 96-hour LL50 value of 3.6 mg/l with rainbow trout (Oncorhynchus mykiss). 

 

The study from Shell (1993a) examined the short-term toxicity of the test substance Hydrocarbons, C10, aromatics, >1% naphthalene, to Oncorhynchus mykiss. Groups of 7 fish were added to each of the control and treatment levels at test initiation, with nominal treatment levels of control, 0.2, 0.7, 2.0, 5.0, 17 and 50 mg/L. The fish were exposed to the test substance for 96h, and fish were observed for mortality at 24, 48, 72, 96 hrs. Water accommodated fractions of hydrocarbons, C10, aromatics, >1% naphthalene, produced a 96-hour LL50 value between 2 and 5 mg/L with rainbow trout (Oncorhynchus mykiss).

 

The study from Exxon (1992) examined the short-term toxicity of the test Hydrocarbons, C10-C13, aromatics, >1% naphthalene, to Oncorhynchus mykiss. Groups of 20 fish were added to each of the control and treatment levels at test initiation, with nominal treatment levels of control, 0.5, 1.0, 5.0, 10.0, and 50 mg/L. The fish were exposed to the test substance for 96h, and fish were observed for mortality at 24, 48, 72, 96 hrs. Water accommodated fractions of hydrocarbons, C10-C13, aromatics, >1% naphthalene, produced a 96-hour LL50 of 3.0 mg/l with rainbow trout (Oncorhynchus mykiss).