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Ecotoxicological information

Toxicity to soil macroorganisms except arthropods

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Reference
Endpoint:
toxicity to soil macroorganisms except arthropods: short-term
Type of information:
experimental study
Adequacy of study:
key study
Study period:
The study was conducted between 04 April 2016 and 15 August 20016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 222 (Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei))
Version / remarks:
2004
Deviations:
no
GLP compliance:
yes (incl. certificate)
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Sponsor Batch No. Ei 2985
- Expiration date of the lot/batch: 01 December 2018

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Ambient 10 - 30”C
Analytical monitoring:
no
Vehicle:
yes
Remarks:
Water
Details on preparation and application of test substrate:

- Method of mixing into soil (if used):
Prior to treatment the initial moisture content of the soil was determined as 25.23% and the maximum water holding capacity (MWHC) as 65.17%. A moisture content equivalent to 55% of MWHC was selected as providing suitable conditions for earthworm development and the soil was pre-moistened to 27.5% MWHC. The volume required to achieve 55% of the maximum water holding capacity was calculated as 233.5 mL water/2200 g dry ISO earthworm soil.
The amount of test item required to achieve the final soil concentration was added as a solution in 233.5 mL of reverse osmosis water to bring the soil moisture content to 55% MWHC. The treated soil was then mixed in for approximately two minutes using a hand held electric mixer.
Prior to treatment the initial moisture content of the soil was determined as 25.23% and the maximum water holding capacity (MWHC) as 65.17%. A moisture content equivalent to 55% of MWHC was selected as providing suitable conditions for earthworm development and the soil was pre-moistened to 27.5% MWHC. The volume required to achieve 55% of the maximum water holding capacity was calculated as 233.5 mL water/2200 g dry ISO earthworm soil.
The amount of test item required to achieve the final soil concentration was added as a solution in 233.5 mL of reverse osmosis water to bring the soil moisture content to 55% MWHC. The treated soil was then mixed in for approximately two minutes using a hand held electric mixer.

- Controls:
For the water control 445.77 mL RO water was mixed into a 4200 g dry weight equivalent of moist soil to bring the soil to 55% moisture content.
For the toxic reference 100 mL of Mascot Systemic treatment solution was mixed into a 4200 g dry weight equivalent of moist soil. A further 345.77 mL of reverse osmosis water was added to bring the soil to 55% of the maximum water holding capacity.

- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Water
- Concentration of vehicle in test medium (stock solution and final test solution): Final = 9.42, 5.234, 2.908, 1.615, 0.8976, 0.4985, 0.277, 0.1538 mg/mL
- Evaporation of vehicle before use: no
Test organisms (species):
Eisenia fetida
Animal group:
annelids
Details on test organisms:
TEST ORGANISM
- Common name: earthworm
- Source: detailed in raw data
- Age at test initiation (mean and range, SD): adult
- Weight at test initiation (mean and range, SD): 250-600 mg

ACCLIMATION
- Acclimation period: 24 hours
- Acclimation conditions (same as test or not): yes
- Health during acclimation (any mortality observed): not specified
Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
no
Total exposure duration:
56 d
Remarks:
On Day 28 of the study the soil was removed from the containers the numbers of live adults were recorded, the soil was returned to the containers and the adult worms discarded. The no. of surviving juvenile worms in each replicate was determined on Day 56
Test temperature:
18 - 22.1 ”C
pH:
6.2
Moisture:
55% Maximum Water Holding Capacity (MWHC)
Details on test conditions:
TEST SYSTEM
- Test container (material, size): plastic; 11 x 17 x 5 cm
- Amount of soil or substrate: 679.2g moist soil
- 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: 8


OTHER TEST CONDITIONS
- Photoperiod: 16h light: 8h dark
- Light intensity: 411 - 800 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :

VEHICLE CONTROL PERFORMED: yes

Nominal and measured concentrations:
9.42, 5.234, 2.908, 1.615, 0.8976, 0.4985, 0.277, 0.1538 mg/mL equivlent to
1000, 555.6, 308.6, 171.5, 95.3, 52.9, 29.4, 16.3 mg/kg dry soil
Reference substance (positive control):
yes
Remarks:
Carbendazim (Mascot Systemic)
Key result
Duration:
28 d
Dose descriptor:
LC50
Effect conc.:
> 1 000 ng/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks:
Adult
Key result
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
> 1 000 ng/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks:
Adult
Key result
Duration:
56 d
Dose descriptor:
NOEC
Effect conc.:
> 1 000 ng/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
not specified
Remarks:
Juveniles
Details on results:
Health and Mortality
Adult mortality was 0% at all rates of application and in both the water control and toxic reference groups.
The LC50 for adult mortality at Day 28 was >1000 mg/kg dry soil. A NOEC of 1000 mg/kg dry soil was achieved.

Bodyweights
There was no statistically significant impact on adult bodyweight at any treatment rate.
The adjusted mean bodyweights on Day 28 at rates of 16.3, 29.4, 52.9, 95.3, 171.5, 308.6, 555.6 and 1000 mg/kg dry soil were 514.1, 483.8, 514.6, 481.5, 501.0, 501.0, 497.7,
453.1 mg respectively compared to 491.0 mg in the water control.
The LC50 for mean bodyweight of the adult earthworm at Day 28 could not be estimated because there was not a dose-response relationship. A NOEC of 1000 mg/kg dry soil was achieved.
There was a statistically significant reduction (p<0.001**) in the bodyweight of the Mascot Systemic group at 5 mg a.i./kg dry soil.

Juvenile Worms
Control group productivity was acceptable (mean of 294.8 juveniles per replicate). The coefficient of variation of the number of juveniles in the negative water control group was 4.98%.
The mean number of juveniles produced at rates of 16.3, 29.4, 52.9, 95.3, 171.5, 308.6, 555.6 and 1000 mg/kg dry soil was 306.0, 321.8, 305.0, 220.8, 251.5, 298.8, 271.3, 275.8 respectively compared to 294.8 in the water control.
There was a significant reduction in the number of juveniles produced at 95.3 mg/kg dry soil.
The EC50 for the number of juveniles on Day 56 could not be estimated because there was not a dose-response relationship. A NOEC of 1000 mg/kg dry soil was achieved.
There was a statistically significant reduction (p<0.001***) in the mean number of juveniles of the Mascot Systemic group at 5 mg a.i./kg dry soil..
Results with reference substance (positive control):
Bodyweights
There was a statistically significant reduction (p<0.001**) in the bodyweight of the Mascot Systemic group at 5 mg a.i./kg dry soil

Juvenile worms
There was a statistically significant reduction (p<0.001***) in the mean number of juveniles of the Mascot Systemic group at 5 mg a.i./kg dry soil.
Validity criteria fulfilled:
yes
Conclusions:
No adult mortality was recorded at any treatment rate.
The LC50 for adult mortality at Day 28 was >1000 mg/kg dry soil. The confidence intervals could not be obtained. A NOEC of 1000 mg/kg dry soil was achieved.
There was no statistically significant impact on adult bodyweight at any treatment rate.
The adjusted mean bodyweights on Day 28 at rates of 16.3, 29.4, 52.9, 95.3, 171.5, 308.6, 555.6 and 1000 mg/kg dry soil were 514.1, 483.8, 514.6, 481.5, 501.0, 501.0, 497.7 and
453.1 mg respectively compared to 491.0 mg in the water control.
The LC50 for mean bodyweight of the adult earthworm at Day 28 could not be estimated because there was not a dose-response relationship. A NOEC of 1000 mg/kg dry soil was achieved.
There was a statistically significant reduction in the number of juveniles produced at 95.3 mg/kg dry soil and in the toxic reference group Mascot systemic.
The mean number of juveniles produced at rates of 16.3, 29.4, 52.9, 95.3, 171.5, 308.6, 555.6 and 1000 mg/kg dry soil was 306.0, 321.8, 305.0, 220.8, 251.5, 298.8, 271.3, 275.8 respectively compared to 294.8 in the water control.
The EC50 for the number of juveniles on Day 56 could not be estimated because there was not a dose-response relationship. A NOEC of 1000 mg/kg dry soil was achieved.
The study was considered valid as there was ≤ 10% adult mortality at four weeks and ≥30 juveniles had been produced in each water control replicate by the end of the test with the coefficient of variation of reproduction ≤ 30%. In addition application of the toxic reference Mascot Systemic at 5 mg a.i./kg dry soil resulted in substantial and unequivocal toxic effects.
Executive summary:

A study was performed to determine the effects of TOFA_TETA_PAA_BADGE_CGE_Adduct (hereafter referred to as TOFA or the test item in the text and as TOFA in tables) on the reproduction and growth of the earthworm, Eisenia fetida, in an artificial soil under laboratory conditions. The method followed was that described in OECD 222 Guideline for the testing of Chemicals, Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei) 2004.

Ten groups of worms were allocated to the reproduction study. Eight groups of 40 worms were treated with the test item at test rates of 16.3, 29.4, 52.9, 95.3, 171.5, 308.6, 555.6 and 1000 mg/kg dry soil weight, mixed into the soil. A similar control group, 80 worms, was maintained in untreated soil to act as a negative control, and a positive control group, 80 worms, was treated with Mascot Systemic (active ingredient carbendazim) at 5 mg a.i./kg dry soil weight. Adult worms were removed from the soil four weeks after treatment and the juvenile worms reared for a further four weeks.

Findings

No adult mortality was recorded at any treatment rate.

The LC50 for adult mortality at Day 28 was >1000 mg/kg dry soil. The confidence intervals could not be obtained. A NOEC of 1000 mg/kg dry soil was achieved. T

here was no statistically significant impact on adult bodyweight at any treatment rate.

The adjusted mean bodyweights on Day 28 at rates of 16.3, 29.4, 52.9, 95.3, 171.5, 308.6, 555.6 and 1000 mg/kg dry soil were 514.1, 483.8, 514.6, 481.5, 501.0, 501.0, 497.7, 453.1 mg respectively compared to 491.0 mg in the water control.

The LC50 for mean bodyweight of the adult earthworm at Day 28 could not be estimated because there was not a dose-response relationship. A NOEC of 1000 mg/kg dry soil was achieved.

There was a statistically significant reduction in the number of juveniles produced at 95.3 mg/kg dry soil and in the toxic reference group Mascot systemic.

The mean number of juveniles produced at rates of 16.3, 29.4, 52.9, 95.3, 171.5, 308.6, 555.6 and 1000 mg/kg dry soil was 306.0, 321.8, 305.0, 220.8, 251.5, 298.8, 271.3, 275.8 respectively compared to 294.8 in the water control.

The EC50 for the number of juveniles on Day 56 could not be estimated because there was not a dose-response relationship. A NOEC of 1000 mg/kg dry soil was achieved.

The study was considered valid as there was ≤ 10% adult mortality at four weeks and ≥30 juveniles had been produced in each water control replicate by the end of the test with the coefficient of variation of reproduction ≤ 30%. In addition application of the toxic reference Mascot Systemic at 5 mg a.i./kg dry soil resulted in substantial and unequivocal toxic effects.

Description of key information

A study was performed to determine the effects of TOFA_TETA_PAA_BADGE_CGE_Adduct (hereafter referred to as TOFA or the test item in the text and as TOFA in tables) on the reproduction and growth of the earthworm, Eisenia fetida, in an artificial soil under laboratory conditions. The method followed was that described in OECD 222 Guideline for the testing of Chemicals, Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei) 2004.

Ten groups of worms were allocated to the reproduction study. Eight groups of 40 worms were treated with the test item at test rates of 16.3, 29.4, 52.9, 95.3, 171.5, 308.6, 555.6 and 1000 mg/kg dry soil weight, mixed into the soil. A similar control group, 80 worms, was maintained in untreated soil to act as a negative control, and a positive control group, 80 worms, was treated with Mascot Systemic (active ingredient carbendazim) at 5 mg a.i./kg dry soil weight. Adult worms were removed from the soil four weeks after treatment and the juvenile worms reared for a further four weeks.

The LC50 for adult mortality at Day 28 was >1000 mg/kg dry soil. The confidence intervals could not be obtained. A NOEC of 1000 mg/kg dry soil was achieved. There was no statistically significant impact on adult bodyweight at any treatment rate.

The LC50 for mean bodyweight of the adult earthworm at Day 28 could not be estimated because there was not a dose-response relationship. A NOEC of 1000 mg/kg dry soil was achieved.

The EC50 for the number of juveniles on Day 56 could not be estimated because there was not a dose-response relationship. A NOEC of 1000 mg/kg dry soil was achieved.

Key value for chemical safety assessment

Long-term EC10, LC10 or NOEC for soil macroorganisms:
1 000 mg/kg soil dw

Additional information