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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:
From 19 January, 2016 to June, 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 222 (Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei))
Deviations:
yes
Remarks:
Temperature in the experiment for 2 h < 18.0°C. Water content in one treatment slightly > 60 % of water holding capacity at study end. However, normal reproduction observed in the controls. Deviations minor and considered uncritical.
GLP compliance:
yes
Analytical monitoring:
not required
Vehicle:
no
Details on preparation and application of test substrate:
For preparation of the nominal treatments the required amounts of the test item for preparation of each treatment 2.2 / 6.8 / 15.9 / 42.0 / 112.6 / 308.7 / 814.4 / 2200.3 mg were each weighted into a glass vessel. To each vessel a part of the deionised water necessary for water content adjustment of the soil was added, it was placed for 10 minutes into an ultrasonic bath and purged into the mixing vessel containing the soil. This purging procedure was performed 3 times for each treatment mentioned above.
Test organisms (species):
Eisenia sp.
Animal group:
nematods
Details on test organisms:
- Species/Strain: Eisenia andrei BOCHÉ
- Age: at least 2 month with clitellum
- Weight: 250 – 599 mg (range in this study)
- Husbandry: Eisenia andrei is bred in the LAUS GmbH throughout the year. The animals are kept for the use in toxicity tests. The keeping is performed following SOP 11501001 (“Hälterung von Eisenia andrei“), in the current edition which is in accordance to OECD Guideline 222.
- Vessels: polyethylene containers with a volume of 20 L
- Medium: horse dung
Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
no
Total exposure duration:
56 d
Test temperature:
17.8 - 22.0°C
Details on test conditions:
TEST SYSTEM
- Test container (material, size):
- Amount of soil or substrate:
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 4
- No. of replicates per control: 8

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
Artificial soil consisting of:

- Sphagnum Peat:
Supplier: Klasman
Batch number: 20140416
10 % of dry matter of artificial soil.
The peat was air dried before use.

- Kaolin Clay
Supplier: Caelo
Batch number: 13276534
20 % of dry matter of artificial soil.
Bolus alba (white clay), pharmaceutical quality.

- Quartz Sand
Supplier: Hagebauzentrum Hotze
Batch number: 20141105
69.5 % of dry matter of artificial soil.
Fine quartz sand (round grains) was used.

- Calcium Carbonate CaCO3
Supplier: Applichem
Batch number: 3C002811

OTHER TEST CONDITIONS
- Photoperiod: 16h
- Light intensity: 800 lux

TEST CONCENTRATIONS
- Test concentrations: 0, 1, 3, 7, 19, 51, 140, 370 and 1000 mg/kg soil dw
Reference substance (positive control):
yes
Remarks:
Benomyl
Key result
Duration:
56 d
Dose descriptor:
NOEC
Effect conc.:
ca. 3 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Key result
Duration:
56 d
Dose descriptor:
EC10
Effect conc.:
ca. 11 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Key result
Duration:
56 d
Dose descriptor:
EC50
Effect conc.:
ca. 115 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
- Mortality at end of exposure period:
Controls:
1 mg/kg soil dw: 2.5%
3 mg/kg soil dw: 0.0%
7 mg/kg soil dw: 0.0%
19 mg/kg soil dw: 0.0%
51 mg/kg soil dw: 0.0%
140 mg/kg soil dw: 0.0%
370 mg/kg soil dw: 0.0%
1000 mg/kg soil dw: 2.5%

- Increase in body weigth of live adults (% of initial weight) at end of exposure period:
Controls: 17%
1 mg/kg soil dw: 31%
3 mg/kg soil dw: 27%
7 mg/kg soil dw: 28%
19 mg/kg soil dw: 31%
51 mg/kg soil dw: 31%
140 mg/kg soil dw: 31%
370 mg/kg soil dw: 28%
1000 mg/kg soil dw: 35%

- No. of offspring produced after 56 d:
Controls: 104
1 mg/kg soil dw: 95
3 mg/kg soil dw: 91
7 mg/kg soil dw: 83
19 mg/kg soil dw: 80
51 mg/kg soil dw: 72
140 mg/kg soil dw: 51
370 mg/kg soil dw: 20
1000 mg/kg soil dw: 7
Validity criteria fulfilled:
yes
Conclusions:
Under the study conditions, the 56 d NOEC (reproduction), EC10 (reproduction) and EC50 (mortality) for the test substance were determined to be 3, 11 and 115 mg/kg soil dw (nominal), respectively.
Executive summary:

A study was conducted to determine the effects of the test substance, DPHA on reproduction of earthworms (Eisenia andrei), according to OECD Guideline 222, in compliance with GLP. The study was performed using eight concentrations ranging from 1 to 1000 mg/kg dry soil. In each replicate, 10 adult animals were exposed to the test item for 28 days in an artificial soil. After 28 days, the living adult animals were counted, and 28 days later, the number of juveniles produced was determined. Six concentrations showed statistically significant inhibition of reproduction ranging from 20 to 93% compared to the control. Adult mortality only occurred in the highest treatment (1000 mg/kg dry soil) and in the blank control. It was 2.5 % each after the first 28 days. The 56 d EC50 of the reference substance Benomyl (tested in a current reference test) was 1.9 mg/kg dry soil, lying within the required range of the guideline. Under the study conditions, the 56 d NOEC (reproduction), EC10 (reproduction) and EC50 (mortality) for the test substance were determined to be 3, 11 and 115 mg/kg soil dw (nominal), respectively (Bangert, 2016).

Description of key information

Based on the study results, the 56 d NOEC (reproduction), EC10 (reproduction) and EC50 (mortality) for the test substance were determined to be 3, 11 and 115 mg/kg soil dw (nominal), respectively.

Key value for chemical safety assessment

Long-term EC10, LC10 or NOEC for soil macroorganisms:
3 mg/kg soil dw

Additional information

A study was conducted to determine the effects of the test substance, DPHA on reproduction of earthworms (Eisenia andrei), according to OECD Guideline 222, in compliance with GLP. The study was performed using eight concentrations ranging from 1 to 1000 mg/kg dry soil. In each replicate, 10 adult animals were exposed to the test item for 28 days in an artificial soil. After 28 days, the living adult animals were counted, and 28 days later, the number of juveniles produced was determined. Six concentrations showed statistically significant inhibition of reproduction ranging from 20 to 93% compared to the control. Adult mortality only occurred in the highest treatment (1000 mg/kg dry soil) and in the blank control. It was 2.5 % each after the first 28 days. The 56 d EC50 of the reference substance Benomyl (tested in a current reference test) was 1.9 mg/kg dry soil, lying within the required range of the guideline. Under the study conditions, the 56 d NOEC (reproduction), EC10 (reproduction) and EC50 (mortality) for the test substance were determined to be 3, 11 and 115 mg/kg soil dw (nominal), respectively (Bangert, 2016).