Registration Dossier

Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2003

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid: viscous

Sampling and analysis

Analytical monitoring:
yes
Remarks:
DOC at test start

Test organisms

Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
Origin: All fish used in the test originated from an own breeding (hatching date: December 02, 2002). Parental fish were supplied by AQUARIUM AM AEGI, Feldstr. 7, D-30159 Hannover.

Holding: Holding was performed at the test facility at 23 ± 2 °C and diffuse light (0.1-10 µmol/m2s, natural photoperiod). The water was changed at least once per week. The dissolved oxygen concentration was more than 80 % of the air saturation value. Only fish from a population with mortality < 5 % within the last 7 days before start of the study were used in the test. No disease treatments were administered throughout holding and testing.

Water: Tap water of local origin was used for holding and testing. The water was filtered on activated charcoal and aerated for at least 24 h to remove chlorine.
Nominal water parameters:
Total hardness: 10 - 250 mg CaCO3/L
pH-value: 6.0 - 8.5
The water is analysed once a year acc. to German tap water regulation.

Feeding: Food was provided 3 times per week. The amount of food was 4 % of the fish body weight per feeding day. The test fish were not fed 24 h before the test was started.

Food: Trouvit 40 I 2; MILKIVIT, D-86666 Borgheim

Number of test fish: 7 zebrafish were used per test concentration and control.

Loading, length of fish: Fish density in the tanks was less than 1 g fish per liter test solution. Average body length at the test start: 2.49 cm
Average body weight at the test start: 0.15 g

Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h

Test conditions

Hardness:
67 mg/L as CaCO3
Test temperature:
23 ± 2 °C
pH:
7.11 - 7.69
Dissolved oxygen:
95-99% saturation
Nominal and measured concentrations:
Test concentrations: 62.5- 125- 250- 500 - 1000 µg/L, prepared with dilution water (nominal)

Stock solution: A stock solution of 10 mg/L was prepared
Details on test conditions:
A static test was performed with one application at the test initiation.

Test duration: 96h

Test vessel: Glass-aquaria loosely covered by glass tops were used.

Test volume: The test volume was about 4 L per vessel.

Aeration: Gentle aeration was provided.

Dilution water: Same as for holding.

Replicates: One replicate per test concentration and control.

Introduction of fish: Fish were introduced randomly to individual replicates.

Feeding of test fish: The fish were not fed during the test.

Water temperature: 23 ± 2 °C

Dissolved oxygen concentration: Not less than 60 % of air saturation value.

Light intensity: 0.1- 10 µmol/m2· s

Photoperiod: A natural photoperiod occurred during the test.

Water quality parameters of pH-value and oxygen-saturation measured after 0, 24, 48, 72 and 96 h were determined to be within the acceptable limits.
Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
354 µg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
LC0
Effect conc.:
250 µg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
500 µg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
250 µg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Details on results:
After 24, 48, 72 and 96 h mortality and behaviour of the test fish was observed.

The test item was clearly dissolved throughout the test.

No symptoms of toxicity were seen up to 250 µg/l. At 500 µg/l one fish showed lethargy before mortality occured.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
In this study the test substance was found to be toxic to zebrafish after 96 h at the nominal concentration level of 500 µg/L and higher.
The LC50 (96 h) was 354 µg/L (330- 379 µg/L), the NOEC after 96 h was the test item concentration 250 µg/L (all nominal concentrations).
Executive summary:

The acute toxicity of the test item to fish (zebrafish) was determined according to OECD-Guideline for Testing of Chemicals No. 203 (1992) from 2003-03-31 to 2003-04-04 at DR.U.NOACK-LABORATORIUM FOR ANGEWANDTE BIOLOGIE, 31157 Sarstedt, Germany.

A static test was conducted with the nominal concentrations 62.5 - 125 - 250 - 500 - 1000 µg/L.

Duration of the test was 96 h. 7 test organisms were exposed to each test concentration and control.

All effect values are based on the nominal concentration of the test item.

The stock solution of the substance was analytically verified via analysis of dissolved organic carbon (DOC, according to DIN guideline 38409 part 3) at test start.

The test substance was found to be toxic to zebrafish after 96 h at the nominal concentration level of 500 µg/L and higher.

The LC50 (96 h) was 354 µg/L (330- 379 µg/L), the NOEC after 96 h was the test item concentration 250 µg/L (all nominal concentrations).

Water quality parameters of pH-value and oxygen-saturation measured after 0, 24, 48, 72 and 96 h were determined to be within the acceptable limits.

The validity criteria of the guideline were met.