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Ecotoxicological information

Toxicity to soil macroorganisms except arthropods

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Administrative data

Endpoint:
toxicity to soil macroorganisms except arthropods: long-term
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Guideline compliant GLP study with good documentation. Klimisch 2 since read across, otherwise Klimisch 1.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 222 (Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei))
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

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

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations:stock solution for 1.3, 13, 26, 52 and 104 mg a.i./kg soil dry weight
- Sampling method: Duplicate specimens from the homogenous stock solution and two specimens of the matrix (deionised water) as control solution were taken from the biological study. The concentration of the test item was determined in one specimen of the stock solution and in one specimen of the control.
- Sample storage conditions before analysis: The samples were stored deep frozen (≤-20 °C) until analysis was performed. Afterwards the samples were again stored deep frozen (≤ -20 °C) and will be kept stored up to the date of the final report.

Test substrate

Vehicle:
no
Details on preparation and application of test substrate:
- Method of mixing into soil (if used):
A stock solution was prepared by weighing 840.2 mg of the test item using an analytical balance and deionised water was added to reach a final net weight of 600.0 g. The resulting solution contained a concentration of 1.40033 mg test item/g. A magnetic stirrer was used to obtain a homogenous solution of the test item.
To reach the target concentration of 160 mg test item/kg soil dry weight (equivalent to 104 mg a.i./kg soil dry weight) 240.0 g of the stock solution were added to the pre-moistened artificial soil corresponding to 2100 g dry artificial soil. The concentration of the added stock solution was 1.4003 mg/g. The achieved nominal concentration was 160.0 mg test item/kg soil dry weight corresponding to 104.0 mg a.i./kg soil dry weight.
To reach the target concentration of 80 mg test item/ kg soil dry weight (equivalent to 52 mg a.i./kg soil dry weight) 120.0 g of the stock solution and 120.0 g water were added to the pre-moistened artificial soil corresponding to 2100 g dry artificial soil. The concentration of the added solution was 0.7002 mg/g. The achieved nominal concentration was 80.0 mg test item/kg soil dry weight corresponding to 52.0 mg a.i./kg soil dry weight.
To reach the target concentration of 40 mg test item/ kg soil dry weight (equivalent to 26 mg a.i./kg soil dry weight) 60.0 g of the stock solution and 180.0 g water were added to the pre-moistened artificial soil corresponding to 2100 g dry artificial soil. The concentration of the added solution was 0.3501 mg/g. The achieved nominal concentration was 40.0 mg test item/kg soil dry weight corresponding to 26.0 mg a.i./kg soil dry weight.
To reach the target concentration of 20 mg test item/ kg soil dry weight (equivalent to 13 mg a.i./kg soil dry weight) 30.0 g of the stock solution and 210.0 g water were added to the pre-moistened artificial soil corresponding to 2100 g dry artificial soil. The concentration of the added solution was 0.1750 mg/g. The achieved nominal concentration was 20.0 mg test item/kg soil dry weight corresponding to 13.0 mg a.i./kg soil dry weight.
To reach the target concentration of 2.0 mg test item/ kg soil dry weight (equivalent to 1.3 mg a.i./kg soil dry weight) a dilution series was prepared. For dilution 1 20.0 g of the stock solution were added to 140.0 g water. For dilution 2 30.0 g of dilution 1 were added to 270.1 g water. 240.0 g of dilution 2 were added to the pre-moistened artificial soil corresponding to 2100 g dry artificial soil. The concentration of the added solution was 0.0175 mg/g. The achieved nominal concentration was 2.00 mg test item/kg soil dry weight corresponding to 1.30 mg a.i./kg soil dry weight.
There were no significant deviations to the nominal target concentration (< 5%). The control was left untreated. While mixing the artificial soil in a laboratory mixer for approximately 5 min the soil of each treatment group (including the control) was moistened with deionised water as described above, the control was moistened with 240.0 g deionised water per batch. Each group was treated in one batch (two in the control) which was then split into the replicates.


- Controls: Untreated soil (moistened with deionised water)

Test organisms

Test organisms (species):
Eisenia fetida
Animal group:
annelids
Details on test organisms:
TEST ORGANISM
- Common name: Eisenia fetida (Savigny 1826)
- Source: Bred under standardised conditions in ibacon laboratories in a breeding medium of cattle manure, peat, sand, calcium carbonate and straw, fed with cattle manure, stored at room temperature.
- Age at test initiation (mean and range, SD): Approximately 7 to 8 months, with well-developed clitellum, age range between test individuals not differing by more than 4 weeks
- Weight at test initiation (mean and range, SD): 300 mg to 587 mg

ACCLIMATION
- Acclimation period: 1 day, in artificial soil
- Acclimation conditions (same as test or not): under test conditions
- Health during acclimation (any mortality observed): healthy worms were used

Study design

Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
no
Total exposure duration:
56 d

Test conditions

Test temperature:
Within the range of 18 °C to 22 °C
pH:
6.3 to 6.5
Moisture:
50.5 to 53.7% of WHC
Details on test conditions:
TEST SYSTEM
- Test container (material, size): Plastic boxes (18.3 cm x 13.6 cm x 6 cm, tapered towards the bottom, with a soil surface of approximately 16.5 cm x 11.5 cm = 189.75 cm²) with perforated transparent lids to enable exchange of air, to minimise evaporation from the artificial soil, and to prevent the worms from escaping.
- Amount of soil or substrate: Each container was filled with 627.3 g of the prepared soil, corresponding to nominal 500 g dry weight.
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 4
- No. of replicates per control: 8
- No. of replicates per vehicle control: no vehicle used

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Geographic location: artificial soil
- Pesticide use history at the collection site: artificial soil
- Collection procedures: artificial soil
- Sampling depth (cm): artificial soil
-Soil preparation:
According to OECD 222:
• 10% Sphagnum-peat, air-dried and finely ground (2 mm with no visible plant remains); (Floragard, Vertriebs GmbH für Gartenbau, 26138 Oldenburg, Germany)
• 20% Kaolin clay (Kaolinite content >30%) (Erbslöh, 65558 Lohrheim, Germany)
• 69.5% fine quartz-sand (F34) containing more than 50% by mass of particle size 0.05 mm to 0.2 mm; (Quarzwerke Frechen, Postfach 1780, 50207 Frechen, Germany)
• 0.5% Calcium carbonate (CaCO3) was added to adjust pH to 6.0 ± 0.5 (Merck, 64293 Darmstadt, Germany).

- Maximum water holding capacity (in % dry weigth): 54%
- Pretreatment of soil: The artificial soil was moistened to approximately half of the final water content 1 day before the application. The additional water required to achieve the final water content was added when applying the test item.


OTHER TEST CONDITIONS
- Photoperiod: 16 h light : 8 h dark
- Light intensity: Within the range of 400 lux to 800 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
mortality and reproduction

VEHICLE CONTROL PERFORMED: no since no vehicle was used

TEST CONCENTRATIONS
- Spacing factor for test concentrations: typically 2 (1.3, 13, 26, 52 and 104 mg a.i./kg soil dry weight)
- Range finding study, performed
- Test concentrations: 1.3, 13, 26, 52 and 104 mg a.i./kg soil dry weight
Nominal and measured concentrations:
Nominal 1.3, 13, 26, 52 and 104 mg a.i./kg soil dry weight, measurement of the stock solution confirmed that the stock solution was correctly dosed ((92 % recovery)

Results and discussion

Effect concentrationsopen allclose all
Duration:
56 d
Dose descriptor:
NOEC
Effect conc.:
>= 104 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
reproduction
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
>= 104 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
mortality
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
>= 104 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
growth
Duration:
56 d
Dose descriptor:
NOEC
Effect conc.:
>= 104 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
food consumption
Details on results:
- Mortality at end of exposure period: none
- Total mass of adults at beginning of test: about 420 mg/worm
- Changes in body weigth of live adults (% of initial weight) at end of exposure period:Control weight after 4 weeks: on average 506.0 mg/worm
- No. of offspring produced: control: 190 worms per container
- No. of unhatched cocoons:not reported
- Morphological abnormalities: none reported
- Behavioural abnormalities: none reported
- Other biological observations: none
Results with reference substance (positive control):
- Results with reference substance valid? yes
- Relevant effect levels: Carbendazim 600 g/L SC NOEC reproduction 1.44 mg/kg dw
Reported statistics and error estimates:
Dunnetts t-test, alpha = 0.05, one-sided smaller

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
earthworm reproduction: NOEC >= 104 mg/kg dw
Executive summary:

In the Klimisch 1 (since read across: Klimisch 2 for this substance) GLP study from Straube (2016) the chronic toxicity of Butanedioic acid, sulfo-, 1,4-diisodecyl ester, sodium salt on earthworms (Eisenia fetida) reproduction and growth in artificial soil was determined in an 56 day test according to OECD 222. The test was performed with Control, 1.3, 13, 26, 52 and 104 mg a.i./kg soil dry weight. The biological results are based on nominal concentrations since the analysis of the stock solution confirmed correct preparation of this solution and the preparation and application of the application solutions was checked and documented by weighing.

Eight replicates with 10 worms each were set up for the control and 4 replicates for each test group. After 28 days 0, 0, 0, 0, 0, 0 and 0% mortality was observed in the control and at nominal test concentrations of 1.3, 13, 26, 52 and 104 mg a.i./kg soil dry weight, respectively.

The reproduction per pre replicate container with 10 adult worms was 190, 165, 171, 150, 170, and 170 living offspring per container (10 worms) control and at nominal test concentrations of 1.3, 13, 26, 52 and 104 mg a.i./kg soil dry weight, respectively.

 

The overall NOEC for reproduction, mortality, growth and food consumption was equal or greater than the highest test concentration, i.e. >= 104 mg/ kg dw.

 

The result will not be considered for the risk assessment.