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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 based on grouping of substances (category approach)
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 222 (Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei))
Version / remarks:
DIN ISO 11268-2: 1998 (E), BBA Guideline (1994)
Deviations:
yes
Remarks:
The room temperature decreased several times to a minimum of 17 °C for a total time of about 17 hours. The soil moisture of all test vessels deviated by more than 10 % of the initial moisture at the end of the test.
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent

Sampling and analysis

Analytical monitoring:
no

Test substrate

Vehicle:
yes
Details on preparation and application of test substrate:
- Method of mixing into soil (if used): The respective test item amount for the limit test item concentration was weighed out, dispersed in acetone (2.5 mL per replicate) and mixed thoroughly with quartz sand (10 g per replicate). After complete evaporation of the solvent the treated quartz sand was given to the artificial soil. Demineralised water was added to adjust the humidity of the artificial soil to a moisture of 56 % of the maximum water holding capacity.
Subsequently, the test medium was thoroughly mixed to ensure a homogenous distribution and 600 g soil dry weight was filled into each test vessel.
- Controls: Artificial soil with addition of quartz sand treated with acetone and moistened with demineralised water without test item was used as solvent control medium.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): organic solvent, acetone
- Concentration of vehicle in test medium (stock solution and final test solution): 2.5 mL per replicate
- Evaporation of vehicle before use: Yes

Test organisms

Test organisms (species):
Eisenia fetida
Animal group:
annelids
Details on test organisms:
TEST ORGANISM
- Common name: Eisenia fetida SAVIGNY 1826, subspecies Eisenia fetida fetida
- Source: Own breeding
- Age at test initiation (mean and range, SD): synchronized earthworm culture, not deviating more than 1 month
- Weight at test initiation (mean): 0.50 g


ACCLIMATION
- Acclimation period: 2 days prior to test start
- Acclimation conditions (same as test or not): same as test
- Health during acclimation (any mortality observed): healthy

Study design

Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
yes
Total exposure duration:
56 d
Remarks:
Removement of adults after 28 days

Test conditions

Test temperature:
17 - 22 °C
pH:
Application rate
[mg test item/kg soil dry weight] pH-value
day 0 day 56
Control 5.96 5.91
Solvent Control 5.89 5.89
1000 5.81 5.82
Moisture:
Application rate
[mg test item/kg soil dry weight] Moisture [%] of dry weight WHCmax of artificial soil
[g/100 g DW]
day 0 day 56
Control 35.4 39.9* 66
Solvent Control 33.8 42.6*
1000 32.9 40.2*
Details on test conditions:
TEST SYSTEM
- Test container (material, size): round plastic boxes, ID: 15 cm, bottom surface area: 177 cm3, height: 14 cm, transparent and perforated lids enabled sufficient gas exchange and light penetration and prevented the test substrate from drying
- Amount of soil or substrate: 600 g DW
- No. of organisms per container (treatment): 10 (40)
- No. of replicates per treatment group: 8
- No. of replicates per control: 8
- No. of replicates per vehicle control: 8


SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Composition (if artificial substrate): 10 % peat, 20 % kaolin, 69 % air dried quartz sand, 0.4 % calcium carbonate
- Organic carbon (%): not determined
- Maximum water holding capacity (in % dry weigth): 66
- CEC: not determined
- Pretreatment of soil: none
- Storage (condition, duration): 4 days after preparation at test conditions in the dark



OTHER TEST CONDITIONS
- Photoperiod: 16 h light - 8 h dark
- Light intensity: 490 +/- 56.8 lx

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Adult mortality, biomass, behaviour and pathological symptoms after 28 days
Reproduction (Number of juveniles) after 56 days

VEHICLE CONTROL PERFORMED: yes


Nominal and measured concentrations:
Nominal application rate: 1000 mg test substance/kg soil dry weight (DW)
Reference substance (positive control):
yes
Remarks:
1.0 - 2.5 - 5.0 mg Carbendazim/ kg soil DW

Results and discussion

Effect concentrations
Duration:
56 d
Dose descriptor:
NOEC
Effect conc.:
1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Remarks:
reduction of body weight, mortality of adult earthworms
Details on results:
- Mortality at end of exposure period: No noteworthy mortality of adult earthworms was observed in the controls as well as at the test item concentration of 1000 mg test item/kg soil dry weight after 28 days of exposure
- Changes in body weigth of live adults (% of initial weight) at end of exposure period: At test start the earthworms had an individual weight of 0.30 - 0.60 g. The biomass of the adult earthworms increased in the controls and at the limit test item concentration of 1000 mg test item/kg soil dry weight over 28 days of exposure with a statistically significant higher increase at 1000 mg test item/kg soil dry weight compared to the pooled control
- No. of offspring produced: The reproduction rate was 395 in the pooled control and 384 at the limit test item concentration of 1000 mg/kg soil dry weight. Compared to the pooled control the reproduction of the earthworms did not statistically significantly differ at the test item concentration of 1000 mg test item/kg soil dry weight
- Behavioural abnormalities: No symptoms or changes in the behaviour of the earthworms were observed in the controls as well as at the limit test item concentration of 1000 mg test item/kg soil dry weight after 28 days of exposure
- Other biological observations: None
Results with reference substance (positive control):
- Results with reference substance valid? Yes
- Relevant effect levels: 2.5 mg Carbendazim/kg soil DW
Reported statistics and error estimates:
t-tests were carried out for the determination of statistically significant differences compared to the pooled control. A Normality Test and an Equal Variance Test were conducted prior to running the t- tests. P-values for both Normality and Equal Variance Test were 0.05. The a-value for ANOVA test (acceptable probability of incorrectly concluding that there is a difference) was a = 0.05.

Any other information on results incl. tables

Mortality of Adult Earthworms in [%] after 28 Days of Exposure

Application rate

[mg test item/kg soil dry weight]

Mortality

[%]

Replicate

1

2

3

4

5

6

7

8

Mean

Control

10

0

0

0

0

0

0

0

1.25

Solvent Control

0

0

0

0

0

10

0

0

1.25

1000

0

0

0

0

0

0

0

0

0

Earthworm Behaviour and Pathological Symptoms after 28 Days of Exposure

Application rate

[mg test item/kg soil dry weight]

Effect

Replicate

1

2

3

4

5

6

7

8

Control

A)

9/10

10/10

10/10

10/10

10/10

10/10

10/10

10/10

H)

1/10

¿

¿

¿

¿

¿

¿

¿

Solvent Control

A)

10/10

10/10

10/10

10/10

10/10

9/10

10/10

10/10

H)

¿

¿

¿

¿

¿

1/10

¿

¿

1000

A)

10/10

10/10

10/10

10/10

10/10

10/10

10/10

10/10

Body Weight Changes of the Adult Earthworms      

Application rate
[mg test item/ kg soil dry weight]

Rep.

Mean body weights per replicate

Mean body weight change of earthworms

Test start

28 days

Per replicate

Mean ± SD

Sign.*

[g]

[g]

[g]

[%]

[g]

[%]

Control

1

0.533

0.768

0.23

44.0

0.22 ± 0.02

43.5 ± 5.22

¿

2

0.501

0.700

0.20

39.7

3

0.513

0.700

0.19

36.5

4

0.487

0.699

0.21

43.5

5

0.500

0.694

0.19

38.8

6

0.502

0.765

0.26

52.4

7

0.481

0.694

0.21

44.3

8

0.470

0.698

0.23

48.5

Solvent Control

1

0.495

0.667

0.17

34.7

0.20 ± 0.04

41.4 ± 8.72

¿

2

0.527

0.694

0.17

31.7

3

0.491

0.687

0.20

39.9

4

0.508

0.737

0.23

45.1

5

0.501

0.674

0.17

34.5

6

0.484

0.747

0.26

54.3

7

0.470

0.722

0.25

53.6

8

0.499

0.686

0.19

37.5

1000

1

0.510

0.728

0.22

42.7

0.24 ± 0.01

48.7 ± 2.73

Yes*

2

0.484

0.736

0.25

52.1

3

0.528

0.786

0.26

48.9

4

0.502

0.741

0.24

47.6

5

0.498

0.745

0.25

49.6

6

0.503

0.754

0.25

49.9

7

0.484

0.722

0.24

49.2

8

0.472

0.708

0.24

50.0

Reproduction Rate (Number of Juveniles after 8 Weeks)

Application rate

[mg test item/kg soil dry weigt]

Rep.

Number of juveniles

Mean ± SD

CV               [%]

[%]

of pooled control

Significance compared to pooled control

Control

1

268

384 ± 54.3

14.1

¿

¿

2

429

3

394

4

394

5

347

6

400

7

401

8

439

Solvent Control

1

497

406 ± 58.2

14.3

¿

¿

2

453

3

421

4

317

5

430

6

365

7

353

8

411

1000

1

478

384 ± 50.9

13.3

97.2

No

2

429

3

355

4

322

5

409

6

349

7

367

8

363

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
Under the conditions of this study, the test item did not induce evident mortality or pathological symptoms of adult earthworms after an exposure to the test limit item concentration of 1000 mg test item/kg soil dry weight for 28 days. Also, the biomass increase of the adult earthworms after 28 days was not statistically significantly reduced compared to the pooled control at the limit test item concentration of 1000 mg test item/kg soil dry weight. After eight weeks of exposure the earthworm reproduction did not statistically significantly differ to the control at the limit test item concentration of 1000 mg test item/kg soil dry weight.

Thus, the No Observed Effect Concentration (NOEC) with respect to mortality, body weight and reproduction of Eisenia fetida fetida after 8 weeks of exposure in artificial soil was determined to be at 1000 mg test item per kg soil dry weight.
Executive summary:

Effects of Pigment Yellow 74on mortality, biomass and the reproductive potential of the earthworm speciesEisenia fetida(Savigny) were determined according to the guidelines DIN ISO 11268-2 (1998), OECD 222 (2004) and BBA (1994)The study was conducted under static conditions over 8 weeks with the limit test item concentration of 1000 mg test item/kg soil dry weight.

The NOEC of the test item concerning mortality, biomass and reproduction of earthworms was determined to be at1000 mg test item/kg soil dry weight.

Summary of All Observed Effects in the Tested Dose 

Effects

[mg test item/kg soil dry weight]

Mortality of adult earthworms

> 1000

Reduction of body weight

> 1000

Reduction of reproduction rate

  (Number of juveniles)

> 1000

Summary of all effects

> 1000

NOEC and LOEC for Reproduction, Mortality and Body Weight

Test Item

Application rate

[mg test item/kg soil dry weight]

NOECreproduction, mortality, body weight

  1000

LOECreproduction, mortality, body weight

> 1000