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Ecotoxicological information

Toxicity to soil macroorganisms except arthropods

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Reference
Endpoint:
toxicity to soil macroorganisms except arthropods: long-term
Type of information:
experimental study
Adequacy of study:
key study
Study period:
16 Dec 2015 - 10 Feb 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP Guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 222 (Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei))
Qualifier:
according to guideline
Guideline:
ISO 11268-2 (Effects of Pollutants on Earthworms. 2. Determination of Effects on Reproduction)
GLP compliance:
yes
Analytical monitoring:
no
Vehicle:
no
Details on preparation and application of test substrate:
- Method of mixing into soil (if used): The test item is poorly soluble in water, therefore, it was applied to the test system by weighing the quantity of the test item necessary to achieve the required test concentra-tion in the soil, to the part of the quartz sand. The sand and test item in the required amounts to attain the selected test concentrations were mixed thoroughly in 0.9-L glass crystallizing dishes (hereafter referred to as test containers). The sand and test item mixture were then mixed thoroughly with the pre-moistened artificial soil to attain the final test mixtures. Milli-Q water was then added, to an amount required to achieve a final moisture content of approximately 50% of the water holding capacity. After mixing the Milli-Q water and the soil, the worms were added to the test mixtures.
- Controls: same treatment method as described above without application of test substance
Test organisms (species):
Eisenia fetida
Animal group:
annelids
Details on test organisms:
TEST ORGANISM
- Common name: earthworm
- Source: ECT Oekotoxikologie GmbH, Floersheim am Main, Germany; Handling and rearing were conducted according to the guideline.
- Age at test initiation (mean and range, SD): adults between 2 months and one year with a well developed clitellum
- Weight at test initiation (mean and range, SD): 306 - 535 mg (average: 410 mg ± 78 mg)

ACCLIMATION
- Acclimation period: 1 d in artificial soil
- Acclimation conditions (same as test or not): same as test
- Health during acclimation (any mortality observed): no mortality observed
Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
no
Total exposure duration:
56 d
Test temperature:
20.0 ± 0.2 °C
pH:
6.2 at test start
6.1 - 6.3 at test termination
Moisture:
50% of WHC (WHC: 65% of DW)
Details on test conditions:
TEST SYSTEM
- Test container (material, size): glass, 0.9 L crystallizing dishes
- Amount of soil or substrate: approx. 520 g soil dw
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 4
- No. of replicates per control: 8
- No. of replicates per positive control: 4
- Other: A sub-set of 8 of the test containers were weighed at the beginning of the test and again every week in order to control the moisture content during the test. If the loss of weight due to evaporation exceeded 10% of the original water content, the evaporated water was replaced by adding Milli-Q water to all test containers.

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Composition (if artificial substrate): 10% sphagnum peat; 20% kaolin clay; 0.4% calcium carbonate; 69.6% air-dried quartz sand
- Maximum water holding capacity (in % dry weigth): 65%

OTHER TEST CONDITIONS
- Photoperiod: 16 h light; 8 h dark
- Light intensity: 500 - 760 lux (7.2 - 10.9 µmol/m²*sec)

EFFECT PARAMETERS MEASURED
- Mortality and growth effects were determined after first 4 weeks of exposure. Any unusual behaviour and morphology was also recorded.
- For the second 4 weeks of exposure the reproduction was assessed. The test containers were placed in a water bath at a temperature of approximately 60 °C. After a period of approximately 30-40 min. the offspring appeared at the substrate surface and could be collected and counted.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.8
Nominal and measured concentrations:
nominal: control, 100, 180, 320, 560 and 1000 mg/kg soil dw.
Reference substance (positive control):
yes
Remarks:
boric acid
Duration:
56 d
Dose descriptor:
NOEC
Effect conc.:
>= 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Duration:
56 d
Dose descriptor:
LOEC
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Duration:
56 d
Dose descriptor:
NOEC
Effect conc.:
>= 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
56 d
Dose descriptor:
LOEC
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
56 d
Dose descriptor:
NOEC
Effect conc.:
>= 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Duration:
56 d
Dose descriptor:
LOEC
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Duration:
56 d
Dose descriptor:
EC10
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: all parameters
Details on results:
- Mortality at end of exposure period:
- Total mass of adults at beginning of test:
- Changes in body weigth of live adults (% of initial weight) at end of exposure period: Weight decreased by 2.6 - 10.3% in the treatments.
- No. of average offspring produced: 80 (control); 75 (100 mg/kg dw); 88 (180 mg/kg dw); 87 (320 mg/kg dw); 99 (560 mg/kg dw); 94 (1000 mg/kg dw)
- Average number of live adults at day 28: 80 (control); 75 (100 mg/kg dw); 88 (180 mg/kg dw); 87 (320 mg/kg dw); 99 (560 mg/kg dw); 94 (1000 mg/kg dw)
Results with reference substance (positive control):
- Results with reference substance valid? yes
- Relevant effect levels: 500 mg/kg dw shows a reduction of reproduction of 98%
Reported statistics and error estimates:
The LOEC/NOEC values were determined by use of the Dunnett’s test. The NOEC values were determined as the highest tested concentration, at which no significant effect was observed compared with the control. The LOEC is the concentration just above the NOEC. The concentrations of test item inhibiting the reproduction 10% and 50% are estimated in accordance with OECD TG 222 and expressed as EC10 and EC50, if possible, by use of the computer program TOXEDO.
Validity criteria fulfilled:
yes

Description of key information

Key value for chemical safety assessment

Additional information

One experimental study is available investigating the chronic effects on soil macroorganisms of Glycerides, C16-18 and C18-unsatd. mono- and di-, citrates (CAS 91052-16-3). The study was performed according to OECD 222 (GLP) with the earthworm Eisenia fetida (Kamper, 2016). Five different concentrations up to 1000 mg/kg soil dw were tested in artificial soil. The mortality was recorded after the first 4 weeks whereas the reproduction was assessed after another 4 weeks of exposure. During exposure no mortality and no effects on reproduction were observed resulting in a NOEC (56 d) ≥ 1000 mg/kg soil dw.