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Ecotoxicological information

Short-term toxicity to fish

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Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2002-03-14 to 2002-03-19
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Well documented guideline study. The substance was identified by a statement from the manufacturer. No analytical monitoring.
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:
other: "Guideline for measuring the acute toxicity of offshore chemicals to juvenile marine fish, the turbot Scophthalmus maximus and the shepshead minnow Cyprinodon variegatus", based on OECD guideline for Testing of Chemicals n°203 (17/07/92)
Deviations:
no
Principles of method if other than guideline:
guideline study
GLP compliance:
yes
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
no
Details on sampling:
Not applicable (no analytical monitoring)
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Water accomodation fractions
WAF of different loadings were prepared in 10 L glass flasks by mixing different amounts of test substance with dilution water (natural seawater with a salinityof 32-34 per thousand filtered on a gravel filter to 50-70µm) for 20 hrs. The mixture was sedimented for minimum 2.5 hrs before the WAF were siphoned into the individual test vessels.
- Controls: Dilution water without test substance
Test organisms (species):
other: Scophthalmus maximus
Details on test organisms:
TEST ORGANISM
- Common name: Turbot
- Source: Stolt Sea Farms, Oye, Norway
- Age at study initiation: Juvenile
- Length at study initiation: 40-60 mm.
- Feeding during test: no feeding

ACCLIMATION
- Acclimation conditions: continuous flow of natural gravel filtered sea water at 10 +/- 3°C and salinity of 32-34 per thousand under normal light intensity (40 W tungsten lamps) at 16:8h L:D cycle.
- Type of food: commercila fish fry feed
- Feeding frequency: every 3 days
Test type:
semi-static
Water media type:
saltwater
Limit test:
no
Total exposure duration:
96 h
Post exposure observation period:
No post exposure observation
Hardness:
Not relevant (seawater study)
Test temperature:
15 +/- 1.5 °C
pH:
7.80 - 8.03
Dissolved oxygen:
85 - 98% saturation
Salinity:
45.0 - 45.5 mS
Nominal and measured concentrations:
Nominal concentrations (based upon loading rate): 27, 66, 164, 410, 1028 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel:
- Type: open
- Material, size, headspace, fill volume: White polyethylene vessels, 35 L, filled with 10 L WAF (test) or dilution seawater (control)
- Aeration: yes
- Renewal rate of test solution: every 48 hrs
- No. of organisms per vessel (test): 7
- No. of organisms per vessel (control): 10
- No. of organisms per vessel (sensitivity control): 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 1
- No. of vessels per sensitivity control (replicates): 1 per concentration
- Biomass loading rate: test 2.91 mg/L; control 1.38 mg/L


TEST MEDIUM / WATER PARAMETERS
- Source of dilution water: Natural seawater with a salinity of 32-34 per thousand filtered in a gravel filter to 50-70 µm.
- Intervals of water quality measurement: every 48 hrs


OTHER TEST CONDITIONS
- Photoperiod: 16 h light / 8 h dark
- Light intensity: 40 W tungsten lamps


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : immobilisation


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.5
Reference substance (positive control):
yes
Remarks:
3,5-dichlorophenol
Duration:
96 h
Dose descriptor:
LL50
Effect conc.:
> 1 028 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
(loading rate)
Basis for effect:
mortality (fish)
Details on results:
- Mortality of control: 0%
- Other adverse effects control: None
Results with reference substance (positive control):
- Results with reference substance valid
- mortality: 20% after 96hr exposure to 0.53 mg/l 3,5-dichlorophenol; 100% mortality after 24hr exposure to 0.67 mg/l 3,5-dichlorophenol
Reported statistics and error estimates:
No calculations possible
Sublethal observations / clinical signs:

Test acceptability:

- Temperature in the exposure vessels during 96 hrs of exposure was 15.0 +/- 0.7 °C.

- The fish in the control series showed normal swimming behaviour showing a cumulative immobilization/mortality of 0% after 96 -hr exposure.

- Saturation of oxygen in the control series and the highest concentration of the test did not fall below 85% during the exposure.

- The fish in the sensitivity control series exposed to 0.53 mg/L of 3.5 -dichlorophenol behaved lethagic but with normal swimming behaviour and a cumulative immobilisation/mortality of 20% after 96 -hr exposure. A second sensitivity control series exosed to 0.67 mg/l of 3,5 -dichlorophenol behaved lethargic after 24 hr exposure with a cumulative immobilization/mortality of 100% after 48 hr exposure.

Validity criteria fulfilled:
yes
Remarks:
-sensitivity of the fish exposed to 0.6 mg/l of 3,5-dichorophenol in the range of 20-80% immobile animals; immobilisation in the control series <= 10% after 96-hr exposure; saturation of oxygen greater than 60%; temperature in the range 15+/-1.5°C
Conclusions:
96h-LL50 > 1028 mg/L (based on nominal loading rate)
Executive summary:

An acute toxicity test was conducted on juveniles of the marine fish Scophthalmus maximus (Turbot). The test substance concentrations (nominal loading rates) ranged from 27 to 1028 mg/L. Test solutions were prepared as individual water accomodated fractions (WAF). Control and dilution water were natural seawater of 32 -34 0/00 salinity filtered through a gravel filter to 50 -70 µm. Test substance was mixed in dilution water for 20 hrs and the mixture was allowed to settle for a minimum of 2.5 hr prior to drawing off the aqueous solution for testing. Test vessels were 35 -L white polyethylene vessels, each containing 10 L of test solution. In the test, 5 concentrations were tested in a geometric series: 27, 66, 165, 410, and 1028 mg/L. Two replicates each containing 7 fishes (biomass loading rate: 2.91 g/L) for the testing and 1 replicate containing 10 fishes (biomass loading rate: 1.38 g/L) for the control were used.

Dissolved oxygen was >60% saturation. The pH was 7.80 - 8.03. The temperature was 15.0 +/- 0.7 °C.

Concentration (mg/L)

Day 0 (initiation)

Day 4 (96 hrs)

Temperature (°C)

Salinity (mS)

% O2

pH

Temperature (°C)

Salinity (mS)

% O2

pH

Control

15.1

45.0

96

8.01

15.7

45.3

87

7.86

27

-

45.0

93

8.01

-

45.4

90

7.82

66

-

45.1

92

8.02

-

45.3

89

7.81

165

-

45.0

91

8.01

-

45.2

90

7.81

410

-

45.0

90

8.03

-

45.4

91

7.82

1028

-

45.0

90

8.04

-

45.5

92

7.83

Test organisms were obtained from a fish farm (Stolt Sea Farms, Oye, Norway). At the test start, the organisms were 40 - 60 mm long juveniles.

No deviations from the protocol were noted.

Analytical monitoring was not performed. Therefore the results are based on the loading rate.

Nominal loading (mg/L)

Number of fish

Initial

Immobilised/dead

24h

48 h

72 h

96 h

Cumulative

Control

10

0

0

0

0

0

27

7 x 2

0

0

0

0

0

66

7 x 2

0

0

0

0

0

165

7 x 2

0

0

0

0

0

410

7 x 2

0

0

0

0

0

1028

7 x 2

0

0

0

0

0

96h-LL50>1028 mg/L (based upon nominal loading rate).

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
January 1986
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
This robust summary has a reliability rating of 2 because the study did not follow GLP and the test containers were open to the air, which allowed for some volatile loss of the test substance. The study followed a standard guideline and was conducted without deviations that would invalidate the study.
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:
other: Unpublished Environment Canada Guideline: Environmental Protection Service 1985; Laboratory for determining the acute lethality of a water-soluble fraction of mineral oil to rainbow trout. Environment Canada, Dartmouth, N.S. June 17.
Deviations:
no
GLP compliance:
no
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
yes
Details on sampling:
Samples for analyses were taken at test initiation and termination. 1-L water samples were siphoned from mid-depth in each container into glass Mason jars. Jars and aluminium foil for capping the jars had been baked overnight at 250 °C. Samples were preserved with 1 mL of 50% HCl. Foil was placed over the tops of the jars and screw caps put on tightly. The samples were kept in the dark in a cold room at approximately 4°C prior to analysis within 2 days.
Vehicle:
no
Details on test solutions:
The treatment solution was prepared by adding one part of the test substance to nine parts dechlorinated tap water for a total volume of 100 L in 120 L cylindrical fiberglass tanks. The tanks were covered with a sheet of polyethylene, and the stock solution was mixed for 20 hours at room temperature with a vortex of approximately 25%. Mixing was performed using a magnetic stir plate and stir bar. After mixing, the solutions were allowed to separate for four hours and the Water Accommodated Fraction (WAF) was then siphoned from the bottom of the mixing vessel and added to the test vessels. Siphons were placed in the tanks prior to adding the test material. Analytical measurements confirmed components of the test material were present in the test solution.
Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
Reported as former name, Salmo gairdneri. Juvenile trout were obtained from Loch Bras d'Or Salmon Farms in Baddeck, Nova Scotia, Canada. The fish had been maintained at a temperature of approximately 5 degrees C at the hatchery. The fish were kept in the testing laboratory in dechlorinated tap water. Prior to the start of the test, the holding water was increased in temperature to approximately 15 degrees C in 2 degrees C/day increments. Fish were fed daily until 24 hours prior to test initiation. The mean length of the fish was 45.9 mm, mean weight was 0.85 gm, and test loading was approximately 0.125 g fish/L. Fish were not fed during the exposure period.

ACCLIMATION
- Acclimation condition: temperature increased from 5°C to 15°C in 2°C daily increments
- Feeding frequency: Daily until 24hrs before the begining of the test
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h
Post exposure observation period:
No post exposure observations
Hardness:
No data
Test temperature:
15.5 - 19.2 °C
pH:
6.7 - 7.3
Dissolved oxygen:
7.6 - 10.5 mg/L
Salinity:
Not relevant (Freshwater study)
Nominal and measured concentrations:
Water accomodated fractions were prepared by adding one part of the test substance to nine parts dechlorinated tap water for a total volume of 100 L in 120 L cylindrical fiberglass tanks. The nominal loading was 788,000 mg/L. The test substance concentration was 200 ppb at test initiation and 120 ppb at termination. The control measured 40 ppb at test initiation.
Details on test conditions:
TEST SYSTEM
- Test vessel:
- Type: open
- Material, size, headspace, fill volume: 60-L circular polyethylene container
- Aeration: None
- No. of organisms per vessel: 10 (loading density: 2.0 L/g of fish/24 h of exposure)
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality at 30 min, 1, 2, 4, 8, 11, and 24 hours, and 2, 3, and 4 days
Reference substance (positive control):
no
Duration:
96 h
Dose descriptor:
LL0
Effect conc.:
87 556 mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
96 h
Dose descriptor:
LL50
Effect conc.:
> 87 556 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Details on results:
No mortality was observed from a water soluble fraction of the test substance prepared at a loading of 87,556 mg/L.
Reported statistics and error estimates:
None were needed as there was no mortality.
Validity criteria fulfilled:
yes
Conclusions:
The water accommodated fraction of the test substance produced a 96-hour LL50 value of >87,556 mg/L and LL0 value of 87,556 mg/L to the fish, Oncorhynchus mykiss. No mortality was observed in either the control or treatment systems.
Executive summary:

The water accommodated fraction of the test substance produced a 96-hour LL50 value of >87,556 mg/L and LL0 value of 87,556 mg/L to the fish, Oncorhynchus mykiss. No mortality was observed in either the control or treatment systems.

Description of key information

The fish were exposed to water accomodated fractions (WAFs) and the results expressed as loading rates. The reported LL50 values range from 0 >1028 mg/L to >70 g/L. No LC50 could be derived from acute short-term studies as they were only based on loading. The risk assessment was performed using the Petrorisk model.

Key value for chemical safety assessment

Additional information

The acute toxicity of Hydrocarbons, C14-C20, aliphatics, ≤2% aromatics was tested on several different species of fish. Four study reports were available and input as endpoint records. Three studies were scientifically reliable according to Klimish (Klimish et al., 1997) scoring (Klimish 1 or 2). The fourth one was scored 4 (unassignable) despite of its scientific value because the test substance was not fully described.

 

Due to the low and variable solubility of the constituents of the substance in water, most of the studies were conducted with water accommodated fractions (WAFs). Marine fish Turbot (reported as Scophthalmus maximus, presently Psetta maxima) was exposed to Hydrocarbons, C14-C18, n-alkanes, isoalkanes, cyclics, <2% aromatics. WAFs and no toxic effect/s (recorded as fish immobilization) was obtained with nominal loadings up to 1028 mg/L (Altin, 2002). Studies conducted with freshwater species (Calmels, 1994; Dufresne 2001a and b) lead to similar results. Zebrafish (reported as Brachydanio rerio, presently Danio rerio) exposure to the same substance as in the above turbot experiment was performed using a mixture of acetone and emulsifier Montanox: no mortality was recorded with nominal concentrations up to 250 mg/L. Dufresne (2001a) conducted a study where freshwater fish fathead minnows Pimephales promelas were exposed to Hydrocarbons, C12-C15,n-alkanes, <2% aromatics and Hydrocarbons, C14-C17,n-alkanes, <2% aromatics as WAFs (1 part of test material + 10 parts of dilution water - exposure to 12.5, 25, 50, 75 and 100% WAF) in a semi-static test with renewal of the test medium every 48h. No significant mortality was reported even with the highest concentration (100% WAF), corresponding to 70 mg/L (nominal loading) for Hydrocarbons, C14-C17, n-alkanes, <2% aromatics.

 

Hydrocarbons, C14-C20, aliphatics, ≤2% aromatics show no acute toxicity towards fish: the toxicity level was never reached in any study despite the high loading level tested. Therefore, these substances are considered to display no acute toxicity to fish.