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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:
2005
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
GLP compliance:
yes
Specific details on test material used for the study:
None
Analytical monitoring:
yes
Details on sampling:
250 µL test medium were sampled in each beaker at 0 and 48 hours, making a total of 1mL medium per treatment per sampling time. Samples were analysed immediately after collection.
Vehicle:
no
Details on test solutions:
80mg of the tested substance was weighed, added to 2000 mL of dilution water and stirred for 67 hours and 20 minutes. Furthermore, ad-hoc dissolution work was performed ultrasonically (about 4×/5 minutes) during the stirring period. Filtering (GF75, 0.3µm) was performed after the conclusion of stirring, and this filtrate was treated as the stock solution. The tested substance concentration of this stock solution was measured and this was treated as the saturation concentration (28.6 mg/L). Dilution water into which the tested substance had not been added was used for the control group. The preparation of the test medium was performed just prior to the start of the trial. Tested concentrations were all prepared by dilution of the stock solution.
Test organisms (species):
Daphnia magna
Details on test organisms:
The testing used Daphnia magna juveniles within 24 hours postpartum. The species was obtained from the National Institute for Environmental Studies, and used after domestication in this testing facility. Visibly healthy and sufficiently large adult females holding juveniles in the subculture were selected, and moved to a separately prepared beaker. When juveniles were obtained from these females, these were put into a new beaker. These juveniles were treated as the parents of the test Daphnia magna, and were domesticated for a 2~4 week period prior to testing. When the juveniles appeared mature and ready to reproduce, the water was replaced at least 2 times a week and the juveniles were removed. After 2 weeks of domestication, the water was replaced prior to the start of the experiment, and the juveniles produced the next day (within 24-hours) were used in the testing (Domestication Lot No.: 050327-1). This lot was free of dead bodies, dormant eggs, and males.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Remarks on exposure duration:
None.
Post exposure observation period:
None.
Hardness:
No data
Test temperature:
19.8°C (constant throughout the test)
pH:
7.4 - 7.7 (min - max)
Dissolved oxygen:
6.2 - 7.8 mg/L
Salinity:
Freshwater.
Conductivity:
No data.
Nominal and measured concentrations:
Nominal concentrations: 0 ; 10 ; 13 ; 16.9 ; 22 ; 28.6 mg/L (the solution were based on a unique stock solution at 28.6 mg/L and a spacing factor of 1.3)
Measured concentrations: 0 ; 11.8 ; 15.3 ; 19.5 ; 26.3 ; 33.6 mg/L
Results were based on nominal concentration because measured concentrations were in the [80-120]% range of the nominal concentrations.
Details on test conditions:
TEST SYSTEM
- Material, size, headspace, fill volume: glass-beakers of 200 mL, filled with 100 mL.
- Aeration: Unknown but oxygen was kept well above the recommended threshold.
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4


OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16L:8D
- Light intensity: Unknown

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Immobilization.

The number of immobile Daphnia magna was recorded at 24 and 48 hours after experiment start. Immobolity was defined as the lack of ability to swim after gentle agitation of the flasks for 15 seconds. Daphnia only able to move along the bottom of the beaker or on the surface of the water were considered as immobile (for water surface, daphnia were considered immobile if and only if the daphnia rose up again after having been forcibly sunk into the water by an operation such as dropping of water droplets. In addition, even in cases of pelagic swimming, daphnia were considered as immobile if they swam in the water not more than 1 time within the 15 seconds.
Reference substance (positive control):
yes
Remarks:
Potassium dichromate
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 28.6 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
mobility
Details on results:
No mortality was recorded in any beaker after 48 hours. Therefore, the 48h-EC50 was considered as superior to 28.6 mg/L.
Results with reference substance (positive control):
Potassium dichromate 48h-EC50 was 0.24 mg/L.
Reported statistics and error estimates:
No immobility was observed in any vessel. Statistics were deemed inapplicable.
Validity criteria fulfilled:
yes
Conclusions:
A GLP test following a protocol equivalent to OECD 202 guideline was conducted on Daphnia magna neonates (< 24 hours old). Tested concentrations were 0 ; 10 ; 13 ; 16.9 ; 22 ; 28.6 mg/L. They were in agreement with measured values at 0 and 48 hours. No immobility was observed in any beaker after 48 hours exposure. Therefore, the 48h-EC50 was considered as superior to 28.6 mg/L.
Executive summary:

A GLP test following a protocol equivalent to OECD 202 guideline was conducted on Daphnia magna neonates (< 24 hours old). Tested concentrations were 0 ; 10 ; 13 ; 16.9 ; 22 ; 28.6 mg/L (spacing factor of 1.3). All groups were tested in quadruplicate (test vessel) and 5 daphnid neonates were introduced by test vessel. The nominal concentrations were in agreement with the concentrations measured at 0 and 48 hours. Immobility was checked every 24 hour by gentle agitation of the test vessels. No immobility was observed in any vessel after 48 hours exposure. Therefore, the 48h-EC50 was considered as superior to 28.6 mg/L.

Description of key information

A GLP test following a protocol equivalent to OECD 202 guideline was conducted on Daphnia magna neonates (< 24 hours old). Tested concentrations were 0 ; 10 ; 13 ; 16.9 ; 22 ; 28.6 mg/L (spacing factor of 1.3). All groups were tested in quadruplicate (test vessel) and 5 daphnid neonates were introduced by test vessel. The nominal concentrations were in agreement with the concentrations measured at 0 and 48 hours. Immobility was checked every 24 hour by gentle agitation of the test vessels. No immobility was observed in any vessel after 48 hours exposure. Therefore, the 48h-EC50 was considered as superior to 28.6 mg/L.

Key value for chemical safety assessment

Additional information