Registration Dossier

Ecotoxicological information

Toxicity to soil macroorganisms except arthropods

Currently viewing:

Administrative data

Link to relevant study record(s)

Reference
Endpoint:
toxicity to soil macroorganisms except arthropods: short-term
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2009-05-04, 2009-06-12
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 207 (Earthworm, Acute Toxicity Tests)
Deviations:
yes
Remarks:
The air temperature in the climate chamber increased and decreased temporarily for short time periods up to 23 °C and down to 17 °C (recommended 20 ± 2 °C. 500 g artificial soil were used instead of 750 g.
GLP compliance:
yes (incl. QA statement)
Analytical monitoring:
no
Vehicle:
no
Details on preparation and application of test substrate:
- Method of mixing into soil: The test item was mixed in an amount of quartz sand sufficient to prepare a mixture for each test item concentration separately. An appropriate amount of these test item/quartz sand mixtures served to prepare the different concentrations of the test item in the artificial soil. The test item was incorporated into the remainder of the artificial soil by thoroughly mixing in the respective test item/quartz sand mixtures for approx. 10 minutes as described in the test guideline.
- Controls: Untreated deionised water control
Test organisms (species):
Eisenia fetida
Animal group:
annelids
Details on test organisms:
TEST ORGANISM
- Common name: Earthworm
- Source: ECT Ökotoxikologie GmbH
- Age at test initiation: adult worms (with clitellum)
- Weight at test initiation: 300 - 600 mg

ACCLIMATION
- Acclimation period: 24 h
- Acclimation conditions: same as test
Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
no
Total exposure duration:
14 d
Post exposure observation period:
None
Test temperature:
17 - 23 °C
pH:
6.0 - 6.2
Moisture:
54.9 - 57.1 % of WHCmax
Details on test conditions:
TEST SYSTEM
- Test container (material, size): 1.5 L preservation jars ("Tulpenform"), Co. Weck, Wehr-Öflingen. The test vessels were covered by glass lids.
- Amount of soil or substrate: 500 g (dw)
- 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)
- Composition (if artificial substrate): 5% Sphagnum peat (no visible plant remains; finely ground and air-dried); 20% Kaolin-Clay (kaolinite content > 30 %); 68 - 69 % Quartz sand (fine sand content with particles between 50 and 200 µm higher than 50 %); ca. 1 % Calcium carbonate (CaCO3)
- Maximum water holding capacity (in % dry weigth): 54.2
- Pretreatment of soil: One day before starting the test the artificial soil (per replicate 500 g dw) was premoistened with deionised water (20 - 40 % of WHCmax)

OTHER TEST CONDITIONS
- Photoperiod: Permanent
- Light intensity: 449 - 558 Lux

EFFECT PARAMETERS MEASURED: Mortality and biomass determined after 7 and 14 days

TEST CONCENTRATIONS: Limit test with two concentrations
- Spacing factor for test concentrations: 10
- Justification for using less concentrations than requested by guideline: Previous Range-Finding Test
Nominal and measured concentrations:
Nominal: 100 and 1000 mg test item/kg soil dw (dry weight)
Reference substance (positive control):
yes
Remarks:
10, 20, 40, and 80 mg chloroacetamide/kg
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:
mortality
Duration:
14 d
Dose descriptor:
LOEC
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.:
100 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Biomass
Duration:
14 d
Dose descriptor:
LOEC
Effect conc.:
1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Biomass
Details on results:
- Mortality at end of exposure period: None, table 1
- Total mass of adults at beginning of test: Table 2
- Changes in body weigth of live adults (% of initial weight) at end of exposure period: Table 1
- Morphological abnormalities: None
- Behavioural abnormalities: None
- Other biological observations: None
Results with reference substance (positive control):
- Results with reference substance valid? Yes
- Relevant effect levels: LC50=50.2 mg/kg
Reported statistics and error estimates:
R/s test procedure confirmed the correspondence with normal distribution. The hypothesis of homogeneity of the biomass data was accepted by the Cochran's test. Therefore, the pairwise comparison of the concentrations with the control test by the Student-t test was used for the statistical analysis.
The ANOVA showed no significant difference but the Student-t test (2-sided, p ≤ 0.05) showed a significant difference in biomass development of individual adults after 14 days between the control and the highest concentrations of the test item tested (i.e. 1000 mg test item/kg soil (dw)) (table 3).
Therefore, the NOEC(Biomass) was determined as 100 mg test item/kg soil (dw) and accordingly, the LOEC(Biomaass) as 1000 mg test item/kg soil (dw).

Table 1. Mean live weight per surviving worm at all concentrations and the control at the beginning of the test, after 7 and 14 days (n = 40). Data are given as absolute values (biomass) and in percent of the initial weight

Concentration

[mg test item/kg soil (dw)]

Mortality

[%]

Biomass

[% of initial weight]

Control 0.0 110.3
100 0.0 104.5
1000 0.0 103.1*

* significantly different to control (Student-t test; p ≤ 0.05, 2-sided)

Table 2. Number and biomass (mg) of the earthworms determined at the beginning, after 7 and 14 days of the test. The concentration is given in mg test item/kg soil (dw) (SD = Standard Deviation; I = Individual Weight).

Day Parameter Control 100 1000


A B C D A B C D A B C D
0 Weight ( l ) 516 306 321 303 460 307 335 386 302 302 336 360
392 371 318 396 444 388 469 325 384 319 401 317
347 336 422 363 436 310 328 328 418 383 397 442
348 303 456 403 338 314 347 377 337 552 407 451
411 307 344 386 391 345 497 410 370 337 320 301
378 328 465 485 387 480 438 369 529 313 485 351
315 397 359 305 381 356 300 413 452 325 358 384
412 362 455 302 321 343 352 346 302 324 359 355
308 325 316 434 342 306 371 370 317 303 334 364
421 310 321 324 316 311 333 312 415 301 375 367
Number 10 10 10 10 10 10 10 10 10 10 10 10
Average 384.8 334.5 377.7 370.1 381.6 346 377 363.6 382.6 345.9 377.2 369.2
SD 61.1 32.1 64.1 62.2 52.2 54.3 66.8 35.1 73.1 76.3 48.3 47.4
7 Weight (Sum) 4040 3800 4222 4156 3931 3870 3926 3861 3999 3738 3985 3549
Number 10 10 10 10 10 10 10 10 10 10 10 10
Average 404 380 422.2 415.6 393.1 387 392.6 386.1 399.9 373.8 398.5 354.9
14
A B C D A B C D A B C D
Weight ( l ) 419 388 350 408 387 360 476 448 388 362 502 333
460 445 494 350 475 386 389 444 515 351 377 333
466 462 535 412 399 355 318 381 389 570 418 321
372 403 393 470 355 518 474 400 393 400 426 296
396 378 370 509 387 337 372 468 358 361 363 394
376 386 494 378 316 318 333 388 436 350 380 321
341 340 422 499 472 383 478 363 390 331 382 463
389 370 369 395 325 352 370 345 251 307 396 441
501 336 356 303 291 419 358 354 362 344 370 379
389 355 372 33 1 360 321 344 326 315 402 365 371
Number 10 10 10 10 10 10 10 10 10 10 10 10
Average 410.9 386.3 415.5 405.5 376.7 374.9 391.7 391.7 379.7 377.8 397.9 365.2
SD 49.9 41.4 67.6 69.7 61.4 59 47.9 47.9 69.5 73.3 42.3 54.9

Table 3. STUDENT t-test for Homogeneous Variances.

Pair-wise comparison of treatments with "Control" by the t-test procedure. Significance was Alpha = 0.05, two-sided; Mean: arithmetic mean; n: sample size; s2: variance; %MDD: minimum detectable difference to Control (in percent of Control); t: sample t; p(t): probability of sample t for Ho: µ1 = µ2; the differences are significant in case p(i) ≤ Alpha ; p(F): probability of F computed by a F-test (Ho: varl = var2 (homogeneity); p(F) > 0.05 is the criterion of variance homogeneity. ((Control(c) and treatment(t) variance was applied; df = N - k; N: sum of treatment replicates n(i); k: number of treatments).

Treatm. [mg/kg] Mean s2 df %MDD t p(t) Sign. p(F)
Control 37.775 112.882





100.00 16.700 249.787 6 61.7 -2.21 0.069 - 0.266
1000.00 11.425 316.129 6 67.1 -2.54 0.044 + 0.210

+ = significant; - = non-significant

Validity criteria fulfilled:
yes
Conclusions:
No toxic effects have been observed in the study with soil living organisms. Statistically significant bodyweight changes were noted in the 1000 mg/kg dw/soil group.
Executive summary:

In this guideline (OECD 207) study conducted with GLP certification, the test material (EC 404-360-3) was determined to not be toxic to soil macroorganisms (worms). The test material (100 mg/kg dw/soil and 1000 mg/kg dw/soil) was exposed to annelid worms (Eisenia fetida) for 14 days. No treatment related mortality was observed during the test. Statistically significant bodyweight changes were noted in the 1000 mg/kg dw/soil group. The NOEC was set at 100 mg/kg dw/soil, as the lowest treated group without effects.

Description of key information

Study conducted to recognised training guidelines with GLP

Key value for chemical safety assessment

Additional information

The acute toxicity of the test item to the earthworm Eisenia fetida was tested in a GLP guideline study according OECD 207 (ECT Oekotoxikologie GmbH 2009). After 14 d no mortality was observed at the control and at the tested limit concentration of 100 and 1000 mg test item/kg artificial soil dry weight. The biomass of individual adults after 14 days amounted to 110.3 % of the initial weight in the control, 104.5 % at 100 mg test item/kg soil (dw) and 103.1 % at 1000 mg test item/kg soil (dw). Therefore, the NOECbiomass was determined as 100 mg test item/kg soil (dw) and accordingly, the LOECBiomass as 1000 mg test itemlkg soil (dw).