Benzamide, N-[[4-[(cyclopropylamino)carbonyl]phenyl]sulfonyl]-2-methoxy-

Toxicity to soil macroorganisms except arthropods

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

toxicity to soil macroorganisms except arthropods: short-term

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

21 Nov - 06 Dec 2005

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 207 (Earthworm, Acute Toxicity Tests)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

no

Vehicle:

no

Details on preparation and application of test substrate:

- Method of mixing into soil (if used): To obtain the nominal test concentrations of 1000, 562, 316, 178 and 100 mg test item/kg dry weight soil, nominal amounts of 500, 281, 158, 89 and 50 mg test item were weighed into a petri dish respectively and quartz sand (washed and calcined) was added up to a final weight of 5 grams for each of the four replicates. Test item and quartz sand were transferred to a mortar and thoroughly mixed with a pestle. Each application mixture (5 g) was transferred separately to artificial soil (500 g dry weight) and mixed thoroughly using a laboratory mixer. Finally 50 mL of deionized water were added and mixed into each replicate to achieve between 40 % and 60 % of the maximum water holding capacity.
- Controls: To the control replicates 5 g of quartz sand (without test item) was mixed into the soil.

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

TEST ORGANISM
- Common name: Earthworm
- Source: Strain of Prof Graff, Forschungsanstalt fur Landwirtschaft, 38104 Braunschweig, Germany. This strain has been held in Bayer CropScience laboratory for several years.
- Age at test initiation: > 2 month
- Weight at test initiation: The average weight was 0.32 g

ACCLIMATION
- Acclimation period: one day
- Acclimation conditions (same as test or not): same as test

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

no

Total exposure duration:

14 d

Test temperature:

20 ± 2 °C

pH:

6.39 - 6.48

Moisture:

water content in the artificial soil (%): 25.9-26.0 (start of study) and 25.6-25.9 (end of study)
WHCmax (mean): 62.2

Details on test conditions:

TEST SYSTEM
- Test container (material, size): 1.5 liter preserving jars
- Amount of soil or substrate: 500 g dry weight test soil (equivalent to 625 g wet weight)
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 4
- No. of replicates per control: 4

OTHER TEST CONDITIONS
- Photoperiod: constant light
- Light intensity: 400 - 800 lux

EFFECT PARAMETERS MEASURED: Seven days after the start of the study, the number of surviving earthworms and after 14 days, the weight, abnormal behaviour, observed symptoms as well as the number of surviving earthworms was determined.

Nominal and measured concentrations:

Nominal test concentrations: 1000, 562, 316, 178 and 100 mg test item/kg dry weight soil

Reference substance (positive control):

yes

Remarks:

Chloroacetamide A.R.

Duration:

14 d

Dose descriptor:

LC50

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

316 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: weight alterations

Results:

Test concentrations (mg test item/ kg dry weight soil)	Container number	Number of surviving worms			Mean weight of Worms (g)
		Day 0	Day 7	Day 14	Day 0	Day 14
Control	1 2 3 4	10 10 10 10	10 10 10 10	10 10 10 10	0.32 0.32 0.32 0.33	0.28 0.31 0.29 0.31
100	1 2 3 4	10 10 10 10	10 10 10 10	10 10 10 10	0.32 0.32 0.32 0.32	0.31 0.28 0.30 0.29
178	1 2 3 4	10 10 10 10	10 10 10 10	10 10 10 10	0.32 0.32 0.31 0.34	0.30 0.27 0.28 0.30
316	1 2 3 4	10 10 10 10	10 10 10 10	10 10 10 10	0.33 0.33 0.33 0.34	0.28 0.28 0.28 0.29
562	1 2 3 4	10 10 10 10	10 10 10 10	10 10 10 10	0.32 0.33 0.32 0.33	0.27 0.28 0.27 0.29
1000	1 2 3 4	10 10 10 10	10 10 10 9	9 10 10 9	0.31 0.33 0.31 0.32	0.27 0.27 0.26 0.28

The LC50 (14d) was > 1000 mg test item/kg dry weight soil. Related to weight alterations and symptoms, the no-observed-effect-concentration (NOEC) was 316 mg test item/kg dry weight soil, the lowest-observed-effect-concentration (LOEC) was 562 mg test item/kg dry weight soil. No morphological and behavioural effects were observed.

Table 1: Validity criteria for OECD 207 (1984)

Criterion	Outcome	Validity criterion fulfilled
Mortality in the control ≤10 % at the end of the test.	Mortality in the control was 0 % at the end of the test.	yes

 

Validity criteria fulfilled:

yes

Remarks:

See Table 1 in "Any other information on results incl. tables".

Reference 2

Endpoint:

toxicity to soil macroorganisms except arthropods: long-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

22 Jun - 22 Aug 2006

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))

Qualifier:

according to guideline

Guideline:

ISO 11268-2 (Effects of Pollutants on Earthworms. 2. Determination of Effects on Reproduction)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

no

Vehicle:

no

Details on preparation and application of test substrate:

- Method of mixing into soil (if used): Appropriate amounts of AE 0001789 were weighed separately for each concentration using an analytical balance and were mixed with 20 g of fine quartz sand. The mixtures were added to artificial soil equivalent to 2030 g dry weight (total dry weight of 2050 g). The control was treated with the same amount of sand as the test item groups.

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

TEST ORGANISM
- Common name: Earthworms
- Source: own bred; Bred under standardised conditions (in a breeding medium of cattle manure, peat, sand and straw, fed with cattle manure, stored at room temperature) by IBACON
- Age at test initiation: ca. 10 months
- Weight at test initiation: 380 mg - 580 mg

ACCLIMATION
- Acclimation period: 2 days, in artificial soil, under test conditions
- Acclimation conditions (same as test or not): same as test condition

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

no

Total exposure duration:

8 wk

Test temperature:

Acclimatisation: 17°C - 22°C
Exposure: 18°C - 21°C

pH:

6.1 to 6.4

Moisture:

Initial soil water content 51.2% to 55.2% of the max. water holding capacity, water content at experimental termination 60.9% to 68.4% of the max. water holding capacity.

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 the soil surface of ca. 16.5 cm x 11.5 cm = 189.75 cm2) with perforated transparent lids.
- Amount of soil or substrate: 659 g (approximately 500 g (dry weight) artificial soil, plus approximately 154 g water, plus approximately 5 g food).
- 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 according to OECD 207

OTHER TEST CONDITIONS
- Photoperiod: 16 h light: 8 h dark
- Light intensity: 420 - 680 lux

EFFECT PARAMETERS MEASURED: Endpoints were mortality, body weight change, feeding activity and reproduction.
Mortality: Number of dead adult earthworms at day 28 after application
Behavioural Abnormalities: Number of damaged adult earthworms (e.g. lack of movement, rigidity) at day 28 after application
Food Given: Cumulative amount of food added to each test container during the test period
Mean Body Weights: Were determined individually at start (day 0) and as mean per test container 28 days after application, using the same washing and weighing procedures as at test initiation
Reproduction: Number of juveniles 8 weeks after application

Nominal and measured concentrations:

Nominal test concentrations: 0 (Control), 0.273, 0.548, 1.09, 2.19 and 4.38 mg test item/kg artificial soil (dry weight) corresponding to 205, 411, 821, 1642 and 3284 g test item/ha equivalent to 199.9, 400.7, 800.5, 1601 and 3202 g a.i./ha.

Reference substance (positive control):

yes

Remarks:

Brabant Carbendazim Flowable

Duration:

8 wk

Dose descriptor:

NOEC

Effect conc.:

>= 4.38 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: mortality, growth, reproduction and feeding activity

Duration:

8 wk

Dose descriptor:

NOEC

Effect conc.:

>= 3 202 g/ha

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: mortality, growth, reproduction and feeding activity

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- Relevant effect levels: The EC50 for reproduction was calculated as 1.14 mg carbendazim/kg soil dry weight.
- Other: The reference item test (dose response) is performed at least once a year as a means of ensuring that the laboratory test conditions are adequate and to verify that the response of the test organisms does not change significantly over time. The GLP conducted experiment was performed from January to March 2006 and is archived under the IBACON Project No. 21343022.

Conclusion:

After exposure of earthworms to AE 0001789 in a chronic test, the NOECfor mortality, growth, reproduction and feeding activitywas determined to be 4.38 mg test item/kg soil dry weight (corresponding to 3202 g a.i./ha), i.e. the highest concentration tested.

Table 1: Validity criteria for OECD 222 (2016)

Criterion from the guideline to be satisfied in the controls	Outcome	Validity criterion fulfilled
Adult mortality over the initial 4 weeks of the test is ≤ 10%.	Adult mortality over the test period was 0%.	yes
Each replicate (containing 10 adults) to have produced ≥ 30 juveniles by the end of the test.	Each replicate (containing 10 adults) have produced ≥ 285 juveniles by the end of the test.	yes
The coefficient of variation of reproduction is ≤ 30%.	The coefficient of variation of reproduction is 11.7%.	yes

 

 

Validity criteria fulfilled:

yes

Remarks:

See Table 1 in "Any other information on results incl. tables".

Description of key information

NOEC (56 d): ≥4.38 mg/kg soil dw (nominal, Eisenia fetida, OECD 222)

Key value for chemical safety assessment

Long-term EC10, LC10 or NOEC for soil macroorganisms:

4.38 mg/kg soil dw

Additional information

The effects of the substance on soil organisms were investigated in an acute 14-day toxicity test on earthworms (Eisenia fetida) according to OECD 207 (M-266322-01-1) and in a long term toxicity test according to OECD 222 (M-277573-01-1). Both studies referred to herein were conducted in accordance with the prevailing OECD testing guidelines under GLP conditions.

The chronic toxicity study was conducted under laboratory conditions using artificial soil as substrate. Earthworms of the species Eisenia fetida were exposed for 56 days to nominal test substance concentrations of 0.273, 0.548, 1.09, 2.19 and 4.38 mg test item/kg soil dw. No effects on mortality, growth, reproduction and feeding activity of the test organisms was observed

The no-observed-effect-concentration (NOEC) was ≥4.38 mg test item/kg soil dry, i.e. the highest concentration tested.

The 14-day LC50 was >1000 mg test item/kg dw as no mortality occurred at this concentration. The NOEC for weight was determined to be 316 mg test item/kg dry weight soil. No morphological and behavioural effects were observed.