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Toxicity to soil macroorganisms except arthropods

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Endpoint:
toxicity to soil macroorganisms except arthropods: short-term
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From October 13, 2003 to November 11, 2003
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 207 (Earthworm, Acute Toxicity Tests)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Specific details on test material used for the study:
- Analytical purity: 49.5% aqueous solution of N-alkyl(C12-16)-N,N-dimethyl–N-benzylammonium chloride (ADBAC) also known as C12-16 alkyl dimethyl ammonium chloride
- Lot/batch No.: DEGE051481
- Storage condition of test material: Room temperature
Analytical monitoring:
no
Vehicle:
yes
Details on preparation and application of test substrate:
Four groups of ten worms were treated with each concentration of test substance. Doses were applied as aqueous solutions and the same volume for all dose levels was used. The control was treated with water only. Worms were rinsed in dechlorinated water, blotted dry and individually weighed at the beginning of the study. At the end of the study they were cleaned in the same manner and weighed in groups of ten.
Test organisms (species):
Eisenia fetida
Animal group:
annelids
Details on test organisms:
Test organisms:
- Common name: Earthworm (Eisenia foetida foetida)
- Source: Huntingdon Life Sciences
- Age at test initiation (mean and range, SD): More than 2 months
- Weight at test initiation (mean and range, SD): 300 - 600 mg
Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
no
Total exposure duration:
14 d
Test temperature:
21.1 - 21.7°C
pH:
5.86
Moisture:
Moisture content was 35% of the dry weight at the beginning and 32-34% at the end of the study.
Details on test conditions:
Test system:
- Test container (material, size): One litre glass container covered with perforated plastic to retain moisture.
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 4
- No. of replicates per control: 4

Source and properties of substrate (if soil):
- Artificial OECD 207 soil, Batch 13/03, (Industrial quartz sand, kaolin clay and sphagnum peat) was used for the main test (pH approximately 5.86)

Other test conditions:
- Photoperiod: continuous light
- Light intensity: 684 lux

Effect parameters measured: Mortality, behavioural and pathological signs

Vehicle control: yes

NB. On Days 7 and 14 the medium was emptied from the containers and worms were removed and counted. On Day 7, the medium was returned to the containers and surviving worms placed on the soil surface.

Nominal and measured concentrations:
Nominal concentrations: 0, 953, 1715, 3086, 5556 or 10000 mg a.i./kg dw of soil
Reference substance (positive control):
yes
Remarks:
Chloroacetamide
Duration:
7 d
Dose descriptor:
LC50
Effect conc.:
7 160 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
mortality
Remarks on result:
other: 95% confidence limits 5560 - 7590 mg a.i./kg dw of soil
Key result
Duration:
14 d
Dose descriptor:
LC50
Effect conc.:
7 070 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
mortality
Remarks on result:
other: 95% confidence limits 5560 - 7400 mg a.i./kg dw of soil
Key result
Duration:
14 d
Dose descriptor:
NOEC
Effect conc.:
953 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
other: body weight
Details on results:
- Mortality at end of exposure period: After 7d, all worms at 10000 and 2 worms at 5556 mg a.i./kg dw of soil were dead. By Day 14, one additional worm died at 5556 mg a.i./kg dw of soil.
- Changes in body weigth of live adults (% of initial weight) at end of exposure period: Group mean body weights were affected by treatment with test substance at 1,715 mg a.i./kg dw of soil and above.
- Morphological abnormalities: No
- Behavioural abnormalities: No
- Other biological observations: At 5556 mg a.i./kg soil dw several live worms were visible at the sides of the containers on Days 1 to 3 then again on Days 8 and 9. At the highest dose level, live worms were visible at the sides and on the surface of the soil in some containers during Days 1 to 6.
Results with reference substance (positive control):
The LC50 values of chloroacetamide to the earthworm, determined in a separate GLP compliant study, were 41 (95% confidence limits 37-43) and 39 (95% confidence limits 35-41) mg a.i./kg dw of soil at Day 7 and 14, respectively. The results obtained in the positive control test were considered acceptable.
Reported statistics and error estimates:
The LC50 values were calculated using a logistic model (Berkson, 1944). 95% confidence limits were estimated by the likelihood ration method (Williams, 1986).

Table 1. Mortality rate (%)

Group No.

Treatment

Initial

Mortality

Total worms

 

(mg a.i./kg dw of soil

No. worms

Day 7

Day 14

as %

1

0

40

0

0

0

2

953

40

0

0

0

3

1715

40

0

0

0

4

3086

40

0

0

0

5

5556

40

2

3

7.5

6

10000

40

40

40

100

 

Table 2. Mean bodyweights (mg)

Group

Treatment

(mg a.i./kg dw of soil

Day 0

Day 14

% decrease

1

0

388

334

14

2

953

389

340

13

3

1715

390

323

17

4

3086

386

304

21

5

5556

389

250

36

6

10000

392

n/d

n/d

 

n/d: no data (as all worms in this group had died)

Validity criteria fulfilled:
yes
Conclusions:
Under the study conditions, the 7d and 14d LC50 values for earthworm (Eisenia foetida) were 7160 and 7070 mg a.i./kg soil dw, respectively and the NOEC was 953 mg a.i./kg soil dw.

Executive summary:

A study was conducted to determine the toxicity to soil macroorganisms of the test substance (49.5% purity%) according to OECD Guideline 207, in compliance with GLP. Six groups of forty earthworm (Eisenia foetida) were allocated to an artificial soil containing 0, 953, 1715, 3086, 5556 or 10000 mg a.i./kg soil dw (nominal concentrations). No analytical dose verification was performed. Mortality was recorded on Days 7 and 14. Worms were weighed at the beginning and end of the study. After 7 days, all worms at 10000 and 2 worms at 5556 mg a.i./kg soil dw were dead. By Day 14, one additional worm died at 5556 mg a.i./kg soil dw. A treatment-related reduction in body weight was observed. Group mean body weights were affected by treatment with test substance at 1715 mg a.i./kg soil dw and above. Under the study conditions, the 7 and 14 d LC50 values were 7160 and 7070 mg a.i./kg soil dw, respectively and the NOEC was 953 mg a.i./kg soil dw (Rodgers, 2004).

Endpoint:
toxicity to soil macroorganisms except arthropods: short-term
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From December 02, 1998 to February 26, 1999
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Qualifier:
according to
Guideline:
OECD Guideline 207 (Earthworm, Acute Toxicity Tests)
Deviations:
yes
Remarks:
analyses were not performed because the substance was readily biodegradable and has a halflife of 40d in soil. It can not be ascertained whether or not the test substance was stable during the test.
GLP compliance:
yes (incl. certificate)
Specific details on test material used for the study:
- Purity: 51.7%
Analytical monitoring:
no
Test organisms (species):
Eisenia fetida
Animal group:
annelids
Details on test organisms:
- Species/strain: Eisenia fetida fetida
- Source of the initial stock: Aquatic Research Organisms INC, Hampton, USA
- Culturing techniques: They were kept in covered plastic vessels (volume 60 L) containing potting compost.
- Age/weight: The weight of the adult earthworms was 0.30 to 0.60 g.
- Pre-treatment: The worms had been acclimatised to artificial soil one d prior to the test.
Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
no
Total exposure duration:
14 d
Test temperature:
18-21°C
pH:
5.56 - 6.06
Moisture:
32.9-35.8%
Details on test conditions:
- Adjustment of pH: No
- Light intensity / photoperiod: Continuous light
- Relevant degradation products: Not determined

Details on test system:
- Artificial soil test substrate: 10% peat (air dried and finely ground), 20% kaolin (kaolinite content 84-87%), 70% quartz sand (sand with particle
size of 100-400 µm) and 0.5% calcium carbonate. PH was 5.73 and moisture content was 20.7%.
- Test mixture: A total weight of 750 g of test substrate was put in each vessel.
- Size, volume and material of test container: Glass dishes of 1.0 l.
- Amount of artificial soil (kg)/ container: 0.750 kg w wt/container
- Nominal levels of test concentrations: 100, 180, 320, 580 and 1000 mg/kg dry weight artificial soil, based product with 51.7% in water
- Active ingredient levels of test concentrations: 52, 93, 165, 300 and 517 mg/kg dry weight artificial soil
- Number of replicates/concentration: 4
- Number of earthworms/test concentration: 40
- Number of earthworms/container: 10
- Light source: 400 to 800 lux
- Test performed in closed vessels due to significant volatility of test substrate: Yes, with perforated plastic film, but not because of volatilisation
but to prevent the test substrate from drying.

The test method was static. The test substance was applied once at test start. The individual live weights of the worms were reported after 14d of exposure. Other effects (pathological symptoms, changes in behaviour of the worms) of the test substance were reported after 7 and 14d of exposure.

Nominal and measured concentrations:
Nominal: 100, 180, 320, 580 and 1000 mg/kg dw of soil
Reference substance (positive control):
yes
Remarks:
2-Chloracetamide (99.5%)
Key result
Duration:
14 d
Dose descriptor:
NOEC
Effect conc.:
1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: weight
Remarks on result:
other: corresponding to >517 mg a.i./kg soil dw
Key result
Duration:
14 d
Dose descriptor:
LC0
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: weight
Remarks on result:
other: corresponding to >517 mg a.i./kg soil dw
Details on results:
LC0 >1000 mg/kg dw
LC50 >1000 mg/kg dw
LC100 >1000 mg/kg dw
No significant mortality and effects were seen in any of the tested concentrations after 14d of exposure. There was no statistically significant difference at the test concentrations 100 - 1000 mglkg compared to the control group. The earthworms in the concentrations 100 to 1000 mg/kg dw showed normal behaviaur and no pathological symptoms. The highest tested concentration without mortality and any other effects was 1000 mg/kg dw.
Results with reference substance (positive control):
Results of the reference test with 2-Chloracetamide showed that the method is sensitive and valid.
Validity criteria fulfilled:
yes
Conclusions:
Under the study conditions, the 14d NOEC in earthworm (Eisenia foetida) was 1000 mg/kg dw (equivalent to 517 mg/kg dw based on a.i.) and the 14d LC0 was more than 1000 mg/kg dw (>517 mg/kg dw based on a.i.).
Executive summary:

A study was conducted to determine the toxicity to soil macroorganisms of the test substance (ca. 51.7% purity) according to OECD Guideline 207, in compliance with GLP. Earthworms (Eisenia foetida) were exposed to a single dose of the test substance at nominal concentrations of 100, 180, 320, 580 or 1,000 mg/kg dw of artificial soil. No analytical dose verification was performed. The individual live weights of the worms were reported after 14 d of exposure. Other effects (pathological symptoms, behaviour of the worms) were reported after 7 and 14 d of exposure. Results of the reference test with 2 -chloracetamide show that the method was sensitive and valid. The substance did not cause a change in behaviour, weight and mortality of the earthworm at any of the tested concentrations after 14 d of exposure. This was probably due to adsorption onto soil. The highest tested concentration without mortality and any other effects was 1000 mg/kg dw. Under the study conditions, the 14 d NOEC in earthworm was 1000 mg/kg dw (equivalent to 517 mg a.i./kg dw) and the 14 d LC0 was > 1000 mg/kg dw (> 517 mg a.i./kg dw) (Noack, 1999).

Description of key information

Based on mortality and effects on bodyweight, the 14 d LC50 and NOEC values in earthworm were determined to be 7070 mg a.i./kg dw and 953 mg a.i./kg dw respectively.

Key value for chemical safety assessment

Short-term EC50 or LC50 for soil macroorganisms:
7 070 mg/kg soil dw
Long-term EC10, LC10 or NOEC for soil macroorganisms:
953 mg/kg soil dw

Additional information

Study 1. A study was conducted to determine the toxicity to soil macroorganisms of the test substance according to OECD Guideline 207, in compliance with GLP. Six groups of forty earthworm (Eisenia foetida) were allocated to an artificial soil containing 0, 953, 1715, 3086, 5556 or 10000 mg a.i./kg soil dw (nominal concentrations). No analytical dose verification was performed. Mortality was recorded on Days 7 and 14. Worms were weighed at the beginning and end of the study. After 7 days, all worms at 10000 and 2 worms at 5556 mg a.i./kg soil dw were dead. By Day 14, one additional worm died at 5556 mg a.i./kg soil dw. A treatment-related reduction in body weight was observed. Group mean body weights were affected by treatment with test substance at 1715 mg a.i./kg soil dw and above. Under the study conditions, the 7 and 14 d LC50 values were 7160 and 7070 mg a.i./kg soil dw, respectively and the NOEC was 953 mg a.i./kg soil dw (Rodgers, 2004).

Study 2. A study was conducted to determine the toxicity to soil macroorganisms of the test substance (ca. 51.7% purity) according to OECD Guideline 207, in compliance with GLP. Earthworms (Eisenia foetida) were exposed to a single dose of the test substance at nominal concentrations of 100, 180, 320, 580 or 1,000 mg/kg dw of artificial soil. No analytical dose verification was performed. The individual live weights of the worms were reported after 14 d of exposure. Other effects (pathological symptoms, behaviour of the worms) were reported after 7 and 14 d of exposure. Results of the reference test with 2 -chloracetamide show that the method was sensitive and valid. The substance did not cause a change in behaviour, weight and mortality of the earthworm at any of the tested concentrations after 14 d of exposure. This was probably due to adsorption onto soil. The highest tested concentration without mortality and any other effects was 1000 mg/kg dw. Under the study conditions, the 14 d NOEC in earthworm was 1000 mg/kg dw (equivalent to 517 mg a.i./kg dw) and the 14 d LC0 was > 1000 mg/kg dw (> 517 mg a.i./kg dw) (Noack, 1999).