Registration Dossier

Ecotoxicological information

Long-term toxicity to aquatic invertebrates

Currently viewing:

Administrative data

Link to relevant study record(s)

Reference
Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 211 (Daphnia magna Reproduction Test)
Version / remarks:
Oct. 2008
Deviations:
yes
Remarks:
(The temperature was slightly higher than stated in the guidelines. The hardness of the test media was not measured. As no decrease of reproduction occurred during the test, these deviations from guideline were stated uncritical.)
Qualifier:
according to
Guideline:
EU Method C.20 (Daphnia magna Reproduction Test)
Version / remarks:
May 2008
Deviations:
yes
Remarks:
(The temperature was slightly higher than stated in the guidelines. The hardness of the test media was not measured. As no decrease of reproduction occurred during the test, these deviations from guideline were stated uncritical.)
GLP compliance:
yes (incl. certificate)
Analytical monitoring:
yes
Details on sampling:
Medium renewal was thrice a week, using freshly prepared test solutions. pH, oxygen and DOC were measured in both the old and the new test solutions. Samples were analysed without prior storage.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: water accommodated fractions (WAFs).
WAFs were prepared by weighing the nominal load, adding the corresponding amount of dilution water (Elendt M4 medium) and shaking vigorously for 24 hours. The resulting solution was filtrated through 0.45 µm filters.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Variety: STRAUS
- Strain/clone: Berlin
- Source: in-house culture originally obtained from the Umweltbundesamt (Federal Environment Agency) in Berlin, Germany in 2007
- Sex: female (reproduction by parthenogenesis)
- Age at study initiation: 0 - 24 hours, not belonging to the first brood of the respective parent animals
- Method of breeding: Similar to the method described in EN ISO 6341:
Vessels: preserving glasses, nominal volume 2 L
Medium: M4-Medium (recipe of ELENDT),
Food: green alga (Desmodesmus subspicatus)
Medium renewal: twice a week
Photo period: 16/8 hours, using neon tubes
Temperature: 20 ± 2 °C
- Feeding during test
Food type: same as for breeding
Amount: 0.15 – 0.20 mg org. carbon/animal/day, double amount before weekend
Frequency: daily

PROCEDURE FOR OBTAINING NEONATE DAPHNIA:
Several days before the start of the test, adult animals having born their first brood were isolated from the stock. 24 hours before the start of the test, the adult animals were separated from the young. 23 hours later, the adults were caught with the help of a glass tube, and the newborn daphnia, aged between 0 and 23 hours, were sieved from the medium and immediately placed into a 250 ml-beaker containing dilution water. After a settling-in period of 30 minutes, animals which showed no apparent damage were used for the test.

ACCLIMATION
- Acclimation period: not relevant, conditions were the same for maintenance and testing
Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
21 d
Hardness:
247 mg/L as CaCO3 (calculated from the composition of the Elendt M4 medium)
Test temperature:
20.6-23.9 °C (slightly higher than stated in the guidelines on several readings. This deviation was stated as uncritical since no decrease of reproduction occurred during the test).
pH:
7.5-8.1
Dissolved oxygen:
8.0-9.9 mg/L air saturation (i.e. > 3 mg/L)
Nominal and measured concentrations:
nominal: loading rate of 100 mg/L
measured: The measured DOC concentrations in the treatment were in a similar range as in the blank control (3.93 mg DOC/L measured at the beginning of the study).
new test solutions: 0.11-3.19 mg DOC/L
old test solutions: 0.18-4.46 mg/DOC/L
Details on test conditions:
TEST SYSTEM
- Test vessel: glass beakers, nominal volume 100 mL
- Aeration: none
- Renewal rate of test solution: Days 3, 5, 7, 10, 12, 14, 17 and 19
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates): 10
- Biomass loading rate: 80 mL/animal

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: deionised water was used for the preparation of the Elendt M4 medium (as stated in the guidelines)
- Dissolved organic carbon (DOC): 3.93 mg/L (measured in the control medium at the beginning of the study)
- Ca/Mg ratio: 3.7:1
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Adjustment of pH: no
-Lighting: ≤ 2000 Lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- The test vessels were inspected for offspring daily. The newborn daphnids were counted and removed from the test vessel. The state of the parental daphnids was recorded (if different from the control). Offspring of dead daphnids were not considered.

RANGE FINDING STUDY
short-term toxicity test to Daphnia:
- Results used to determine the conditions for the definitive study (based on loading rates): 48-h NOELR ≥ 100 mg/L, 48-h EL50 >100 mg/L
Reference substance (positive control):
not required
Key result
Duration:
21 d
Dose descriptor:
NOELR
Effect conc.:
>= 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Remarks on result:
other: results are expressed in terms of loading rates
Key result
Duration:
21 d
Dose descriptor:
EL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Remarks on result:
other: results are expressed in terms of loading rates
Key result
Duration:
21 d
Dose descriptor:
LL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks:
(parental)
Remarks on result:
other: results are expressed in terms of loading rates
Details on results:
- Type and number of morphological abnormalities: none reported
- Type and number of behavioural abnormalities: none reported
- Mortality of parent animals: control 0%; treatment group 10% (1/10)
- Time to first brood release: 10 days for the control and the treatment group
- Mean number of offspring produced per parent animal in the control: 60 (coefficient of variation: 7.2%; n=10)
- Mean number of offspring produced per parent animal in the treatment group: 61 (coefficient of variation: 13.8%; n=9)
Validity criteria fulfilled:
yes
Remarks:
mortality of the parent animals was 0% at the end of the test (≤20%), mean surviving offspring per parent animal was 60.4 (≥60) and the coefficient of variation was 7.2% (<25%)

Description of key information

21-d NOELR (based on loading rates) ≥100 mg/L for Daphnia magna (OECD 211, EU C.20)

Key value for chemical safety assessment

EC10, LC10 or NOEC for freshwater invertebrates:
100 mg/L

Additional information

The NOEC for the test material is much higher than its water solubility.