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Ecotoxicological information

Short-term toxicity to fish

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Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
28 Oct. 2008 - 27 Jan. 2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP Guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009
Report Date:
2009

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): anthracene oil (> 50 ppm BaP)
- Molecular formula (if other than submission substance): not applicable
- Molecular weight (if other than submission substance): not applicable
- Substance type: organic
- Physical state: liquid
- Stability under test conditions: WAF (water accommodated fraction) limited
- Storage condition of test material: room temperature, exclusion of light

Sampling and analysis

Analytical monitoring:
yes

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Water Accommodated Fractions (WAFs) prepared by stirring various amounts of the test item in dechlorinated tap water
for 24 h with magnetic stirrers. The WAFs were prepared in 20-L glass fish tanks.
These were equipped with a glass tube that allowed the separation of the water phase by siphoning.
The test loadings of anthracene oil were weighed on watch-glasses, that afterwards were placed into the glass vessels.
The glass tanks were then filled with the tap water.
Mixing was carried out at a speed that was slow enough not to cause dispersion or emulsification of the undissolved fraction
of the test item. To ensure this, the vortex developed at the surface by stirring was set at ~ 10 % of the water depth.
After stirring for 24 h the WAFs were allowed to stand for 1 h before use to facilitate phase separation.
The extracts gained with this method were clear.

- Differential loading: control and 100 mg/L
- Controls: without test material (blank; tap water)
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): none
- Evidence of undissolved material (e.g. precipitate, surface film, etc): no

Test organisms

Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: Zebra fish
- Strain:
- Source: Max-Planck-Institute für Entwicklungsbiologie, Tübingen
- Age at study initiation (mean and range, SD): no data
- Length at study initiation (length definition, mean, range and SD): 3.0 - 3.5 cm
- Weight at study initiation (mean and range, SD): no data
- Feeding during test: none


ACCLIMATION
- Acclimation period: >12 days
- Acclimation conditions: same as test
- Type and amount of food: no data
- Feeding frequency: no data
- Health during acclimation (any mortality observed): mortality <1%

Study design

Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Remarks on exposure duration:
water/WAF renewal: every 24 h
Post exposure observation period:
none

Test conditions

Hardness:
Total hardness 8.97 °dH
Carbonate hardness 6.98 °dH
Test temperature:
21.1 - 22.0 °C (23 +-2 °C)
pH:
8.2 - 8.5
Dissolved oxygen:
81 - 96 % of saturation
Salinity:
no data
Nominal and measured concentrations:
nominal: 100 mg/L
measured (as TOC):

Period Control 100 mg/L (loading)
===================================
0-24 h start 3.0 2.6 mg/L (TOC of WAF)
0-24 h end 2.8 2.4 mg/L (TOC of WAF)
72-96 h start 2.7 2.4 mg/L (TOC of WAF)
72-96 h end 2.7 2.5 mg/L (TOC of WAF)
=====================================
Note: The oil-related TOC cannot be estimated from these data, as the control values are extraordinarily high.
Details on test conditions:
TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: 20 L, test volume 7 L
- Aeration: none
- Renewal rate of test solution (frequency/flow rate): every 24 h
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 1
- Biomass loading rate: 1 fish/L


OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: light / dark: 14 h/ 10 h
- Light intensity: fluorescent tubes, no data


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Swarming-behaviour, activity, balance, surfacing, mortality, breathing frequency, apathy


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2 - 2.5
- Range finding study
- Test concentrations: based on previous daphnia study
Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Duration:
96 h
Dose descriptor:
LL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: ; (LL = lethal loading)
Duration:
96 h
Dose descriptor:
NOELR
Effect conc.:
< 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Details on results:
No mortality

- Control: Swarming behaviour and activity were normal.
- 100 mg/L: Activity was slightly reduced, and surfacing could be observed at the start of the observation periods
0 – 24 h, 24 – 48 h, 48 – 72 h and 72 – 96 h. These sublethal effects could not be observed at the end of
the observation periods 0 – 24 h, 24 – 48 h and 48 – 72 h.
However slightly reduced activity could also be observed at the end of the observation period 72 – 96 h.

Observations at the end of the test after feeding of the fish:

- Control: All fish fed immediately.
- 100 mg/L: All fish fed with slight retardation compared to the control.

After the end of the test, the fish were kept in de-chlorinated tap water from 20.12.08 to 23.12.08.
During this period, fish derived from the control and the test loading rate of 100 mg/L showed normal behaviour.

Any other information on results incl. tables

Number of dead fish and cumulative lethality [%] at various observation times (Report, Tab. 3)

Time [h]

Control

100 mg/L

Number

[%]

Number

[%]

24

0/7

0

0/7

0

48

0/7

0

0/7

0

72

0/7

0

0/7

0

96

0/7

0

0/7

0

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Remarks:
- Dissolved oxygen in all vessels: >= 81 % (nominal value > 60 %) - Lethality in the control: 0 fish (nominal value ≤ 1 fish)