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Long-term toxicity to fish

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Reference
Endpoint:
fish early-life stage toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Apr - July 2006
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 210 (Fish, Early-Life Stage Toxicity Test)
Qualifier:
according to guideline
Guideline:
EPA OPPTS 850.1400 (Fish Early-life Stage Toxicity Test)
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
During the prolonged toxicity study, the following sampling regime was applied for confirmation of test concentrations.
Number Fresh solutions: Duplicate samples from each solution freshly prepared.
spent solutions: Singular samples from each vessel, i.e. six samples per group.
Frequency Fresh solutions on days 0, 4, 9, 14, 21 and 28; spent solutions from each vessel on days 7, 11, 16, 23 and 30.
Volume 2 ml
Vehicle:
yes
Details on test solutions:
Preparation of stock and test solutions
The standard test procedures required generation of test solutions, which should contain completely dissolved test substance concentrations or stable and homogeneous mixtures or dispersions. The testing of concentrations that disturb the test system should be prevented (e.g.
precipitation or a film of the test substance on the water surface).
The batch of VAA489 VAL tested was a white powder with a purity of 99.0% (HPLC) and completely soluble in test medium at the concentration tested.
Preparation of test solutions started with quantitative addition of test substance to test medium at 10 mg/l. To accelerate the dissolving of the test substance in the test medium the solutions were treated with ultrasonic waves in two steps of approximately 20 and 10 minutes, respectively.
The final test solutions were all clear and colourless.
Test organisms (species):
Pimephales promelas
Details on test organisms:
Fathead minnow (Pimephales promelas, Teleostei Cyprinidae) Rafinesque.
Test type:
semi-static
Water media type:
freshwater
Total exposure duration:
30 d
Hardness:
180 mg/l expressed as CaC03 (original hardness of ISO-medium is 250 mg/l).
Test temperature:
27°C
pH:
at 7.8 ± 0.2
Nominal and measured concentrations:
The analytical support confirmed the repeatability of the preparation of the test concentrations and that the concentrations measured remained stable during the periods between renewals. Concentrations measured were in the range of 100 % (92104 %) of the nominal value of 10 mg/l in both fresh and spent (48 or 72-h old) solutions.
Details on test conditions:
The batch of VAA489 VAL tested was a white powder with a purity of 99.0% (HPLC) and completely soluble in test medium at the limit concentration of 10 mg/l tested. The test includeda blank control and started by placing fertilised eggs in petri-dishes under semi-static conditions until all eggs had hatched (day 4). Then the hatched larvae were transferred to 1 -titer all glass vessels and maintained under semi-static conditions until day 25 when they were transferred too-liter al glass vessels. During the embryonic and larval phases the fish were observed for effects on development, appearance and swimming behaviour. At the end the fish were measured and weighed. Samples for chemical analysis of the actual VAA489 VAL concentration were taken from various intervals during the study of both fresh and spent solutions.
The test starts by placing fertilised eggs in petri-dishes under semi-static conditions and is continued for a total period of 30 days. Lethal and sub-lethal effects are assessed and compared with control values to determine the lowest observed effect concentration, the no observed effect concentration and the maximum acceptable toxicant concentration.
Test type Semi-static with renewal of the test solutions three times a week (generally on Monday, Wednesday and Friday).
Test duration 30 days
Introduction egg 2-4 hours after fertilisation, all embryos in early blastula stage.
Test vessels Sterile petri-dishes (diameter: 15 cm) until day 4 when all eggs had hatched and the larvae were transferred to allglass vessels of ca. 1 liter. These test vessels were covered with a petri-dish lid.
Experimental design The experiment (nominal day 0) started with 120 fresh and-healthy fertilised fathead. minnow eggs per test group. The fertilised eggs were randomly distributed and divided equally over six petri-dishes. Each petri-dish contained 20 eggs in 100 ml test medium and wereincubated under gently continuous shaking. On day 4 the hatched larvae were transferred to- all-glass vessels of ca. 1 liter. The test vessels were protected against unwanted disturbance by performing the test in a climate room with no entry for unauthorised persons
Reference substance (positive control):
no
Key result
Duration:
30 d
Dose descriptor:
LOEC
Effect conc.:
> 10 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Key result
Duration:
30 d
Dose descriptor:
NOEC
Effect conc.:
10 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
The present study assessed the possible lethal and sub-lethal effects of VAA489 VAL during the embryonic and early larval development of the fathead minnow. The results led to the following conclusions for VAA489 VAL:
1. VAA489 VAL did not induce any significant, visible effects on the development of fathead minnow embryos at a limit concentration of 10mg.
2. VAA489 VAL did not significantly affect time of hatching or the hatching success at a limit concentration of 10 mg/l.
3. VAA489 VAL did not affect survival, growth or development of fish larvae during the whole test period at a limit concentration of 10 mg/l.
Hence, the NOEC of VAA489 VAL for the early life stages of fish is 10 mg/l, the regulatory limit concentration. The overall LOEC is > 10 mg/l.
Validity criteria fulfilled:
yes
Conclusions:
VAA489 VAL did not induce any significant, visible effects on the development of fathead minnow embryos or significantly affect time of hatching or the hatching success at the limit concentration of 10 mg/l. Further, VAA489 VAL did not affect survival, growth or development of the larvae at the limit concentration of 10 mg/l. Post-hatch survival in the controls and the VAA489 VAL treated replicates was > 80%.
In conclusion: the NOEC of VAA489 VAL for the early life stages of fish was 10 mg/l, the regulatory limit concentration. The overall LOEC is > 10 mg/l.

Description of key information

The NOEC of VAA489 VAL for the early life stages of fish was 10 mg/l, the regulatory limit concentration. The overall LOEC is > 10 mg/l.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Effect concentration:
10 mg/L

Additional information