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Short-term toxicity to aquatic invertebrates

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Reference
Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
from 2008-08-04 to 2008-09-07
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Well documented guideline study. NOELR according to OSPAR COM (not relevant in the case of acute toxicity test according to EC standards). Based on WAFs. No analytical monitoring.
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:
other: ISO 14669 - 1999 Water quality - Determination of acute lethal toxicity to marine copepods (Copepoda, Crustacea)
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
- Method: Water accomodation fractions (WAF) were prepared individually by stirring appropriate amounts of the test substance in natural seawater according to ISO 14442-2006. The solutions were stirred with a spin bar for 22 hrs at a speed which formed a vortex one-third of the depth of the fluid content, followed by standstill for 1.5 hrs. Closed aspirator bottles were used and samples for testing were drawn off through a drain port near the bottom of the bottle.
For two of the concentrations, 3.0 and 99.2 mg/L WAF the spin bar had stopped during the night. They were turned again for 1 hour in the morning. It is not known for how long the spin bars stood still.

-Seawater: natural seawater from an unpolluted site at about 80 m depth in Byfjord, collected from a tap at Akvamiljo on 11.06.2008, after filtration through a sand filter. This seawater had a salinity of 34.8 per thousand. At the lab, the water was filtered through a GF/C filter and briefly heated to 75°C prior to use. In this test seawater with addition of nutrients (according to ISO 110253 - 2006) was used since an algae test was also conducted.
Test organisms (species):
other aquatic arthropod: Acartia tonsa
Details on test organisms:
TEST ORGANISM
Acartia tonsa, Marine planktonic copepod

- Source: bought as eggs from Marin Biologiisk laboratorium, Kobenhaven University, Helsingor.
- Age at study initiation: 15 days
- Feeding during test: no feeding

ACCLIMATION
The test organisms used in the test were hatched 21.07.2008 at M. Lab and held at a temperature of 19.5 - 24.1 °C. They were fed with a culture of Rhodomonas baltica from day 2.
Test type:
static
Water media type:
saltwater
Limit test:
no
Total exposure duration:
48 h
Post exposure observation period:
No post exposure observation
Hardness:
Not relevant (seawater study)
Test temperature:
19.5 - 21.5 °C
pH:
8.3
Dissolved oxygen:
7.0 - 7.4 mg/L
Salinity:
34.8 per thousand
Nominal and measured concentrations:
Nominal concentrations (loadings): 3.0, 10.0, 31.7, 99.2, 319, 1038, and 3193 mg/L (WAF)
Details on test conditions:
TEST SYSTEM
- Test vessel:
- Type: open
- Material, size, headspace, fill volume: 28 mL glass rolled rim vials each containig 25 mL of test solution
- Aeration: none
- Renewal rate of test solution (frequency/flow rate): no renewal (static test)
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 6


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: natural seawater from an unpolluted site at about 80 m depth in Byfjord,filtered through a sand filter, then through a GF/C filter and briefly heated to about 75°.


OTHER TEST CONDITIONS
- Photoperiod: 16 hrs light / 8 hrs dark


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : lethality after 24 and 48 hrs


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 3.20 (mean)
Reference substance (positive control):
yes
Remarks:
3,5-dichlorophenol
Duration:
24 h
Dose descriptor:
LL50
Effect conc.:
> 3 193 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
(loading)
Basis for effect:
mortality
Remarks on result:
other: WAF
Duration:
48 h
Dose descriptor:
LL50
Effect conc.:
> 3 193 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
(loading)
Basis for effect:
mortality
Remarks on result:
other: WAF
Duration:
24 h
Dose descriptor:
other: LL90
Effect conc.:
> 3 193 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
(loading)
Basis for effect:
mortality
Remarks on result:
other: WAF
Duration:
48 h
Dose descriptor:
other: LL90
Effect conc.:
> 3 193 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
(loading)
Basis for effect:
mortality
Remarks on result:
other: WAF
Details on results:
- Mortality of control: 10%
Results with reference substance (positive control):
- Mortality: 66.7%

Validity of the test: the acceptance criteria were fulfilled

- All copepods were from the same population

- The percentage mortality in the blank did not exceed 10%

- The 48-hr mortality obtained after exposure to 1.0 mg/L 3,5 -dichlorophenol was 66.7 % (in the expected range of 20%-80%)

- The oxygen concentration was greater than 4 mg/L throughout the test (7.0 - 7.4 mg/L)

Validity criteria fulfilled:
yes
Remarks:
Copepods all from the same population ; mortality in blanks<=10%; oxygen concentration>4mg/l; 48-hr mortality with reference substance within 20-80%
Conclusions:
48-hr LL50 > 3193 mg/l (based upon nominal loading rate)
Executive summary:

An acute toxicity test was conducted on the marine copepod Acartia tonsa. The test substance concentrations (nominal loading rates) ranged from 32.9 to 3190 mg/L. Test solutions were prepared as individual water accomodation fractions (WAF). Control and dilution water were natural seawater from an unpolluted site at about 80m depth in Byfjord, filtered through sand filter at the collection, then through a GF/C filter and briefly heated to 75°C prior to use. Test substance was mixed in dilution water for 22 hours, and the mixture was allowed to settle for 1.5 hours prior to drawing off the aqueous solutions for testing. Test vessels were glass 28 mL rolled rim vials, each containing 25 mL of test solution. In the test, 5 concentrations were tested in a geometric series: 32.9, 99.3, 320, 1000, and 3190 mg/L. Four replicates for the test substance and six for the blank were used, with 5 copepods per vial.

Dissolved oxygen was >4 mg/L during the study. The pH was 8.1 - 8.2. The temperature was 20.3 - 21.9 °C.

Nominal loading

(mg/L)

Dissolved oxygen

(mg/L)

pH

Control

7.4

8.3

3.0

7.1

8.3

10.0

7.2

8.3

31.7

7.2

8.3

99.2

7.1

8.3

319

7.1

8.3

 1038  7.1  8.3
 3193  7.1  8.3

Test organisms were obtained from eggs. At test start, the test organisms were 15 days (after hatching).

No excursions from protocol were noted.

Analytical monitoring was not performed.

Nominal Loading

(mg/L)

Total number immobilized

(24 hrs)

(48 hrs)

Control

1

3

3.0

0

3

10.0

0

2

31.7

1

6

99.2

0

7

319

0

2

 1038  0  4
 3193  0  3

48-hr LL50 > 3193 mg/L (based upon nominal rate)

Description of key information

The test substance is poorly soluble and made of constituents with various water solubility. As a consequence, the exposures were performed

with Water Accomodated Fractions (WAFs). Therefore, the results are based on nominal loadings and the LC50 value given below is actually a LL50 (lethal loading) value. No LC50 could be derived from the acute toxicity studies.

The 48h-LL50 is > 3000 mg/l.

Key value for chemical safety assessment

Additional information

Eleven study reports were available and input as endpoint summary records. All the studies investigate the toxicity of the substance “Hydrocarbons, C14-C18, n-alkanes, isoalkanes, cyclics, ≤ 2% aromatics” on a marine copepod, Acartia tonsa exposed to water accommodated fractions (WAFs). Several studies indicate very high values (up to 69 155 mg in Altin, 1998b), In most of the studies the calculation of LL50 could not be achieved because of the low mortality levels obtained (Aunaas, 1996a and c; Larsen, 2006a, b, d and e; 2008a-c). When more than 5 values are available for LL50, the geometric mean of all the values is considered for LL50. However, in this particular case, as a great number of studies could not achieve the calculation of LL50, the geometric mean cannot be calculated, but 9 studies among 11 found that LL50 was greater than 3000 mg/l. Therefore, it is considered that LL50 is >3000 mg/l.