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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:
07 Dec 2015 - 08 Jan 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Qualifier:
according to guideline
Guideline:
other: The Guidelines for Testing of Chemicals - Effects on Biotic Systems
Version / remarks:
2013
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: SEPA 2004, HJ/T 153-2004, the guidelines for the testing of chemicals.
Version / remarks:
2004
Deviations:
no
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 207 (Earthworm, Acute Toxicity Tests)
Version / remarks:
1984
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
no
Vehicle:
no
Details on preparation and application of test substrate:
- Method of preparation of test substrate: Direct additions into the soil. A total amount of 10 g mixture of the test substance and quartz sand was prepared firstly and then mixed with 550 g dry weight of soil for each test container. The medium was thoroughly mixed and deionised water was added finally to give an overall moisture content of about 35 per cent of the dry weight. Moisture content was determined by weighing and then drying a small sample at 105 °C and re-weighing.
- Controls: yes, blank control
Test organisms (species):
Eisenia fetida
Animal group:
annelids
Details on test organisms:
TEST ORGANISM
- Common name: earthworm
- Source: Nanjing Earthworm Nursery, Nanjing, China
- Age at test initiation: adult (two months old with clitellum)
- Weight at test initiation: 376 - 416 mg

ACCLIMATION
- Quarantaine period: Earthworms newly obtained in the lab were released from quarantine after 14 days and then this batch was pre-cultured in soil with cow dung for 272 days (Batch No.: E20150327E).
- Acclimation period: Before the test, the worms were acclimated in test soil for another 24 h. The worms were then washed quickly with distilled water and surplus water absorbed on filter paper before use.
Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
yes
Total exposure duration:
14 d
Test temperature:
19.8 - 20.3 °C
pH:
Start: 6.14 - 6.33
End: 6.17 - 6.36
Moisture:
Moisture of soil at start: 35%
Moisture of soil at end: 27 - 30%
Air humidity: 78 - 82%
Details on test conditions:
TEST SYSTEM
- Test container: 1L glass spoutless beakers, 12 cm diameter, covered with plastic films with small ventilation holes
- Amount of soil or substrate: 560 g (dry weight)
- 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
- Composition (artificial substrate): 10% Sphagnum peat (pH 6.0, no visible plant remains, finely ground, dried), 20% kaolin clay (kaolinite content preferably above 30%), 70% quartz sand (fine sand should be dominant with 80% of the particles between 50 and 200 microns).
- Moisture content: 35% dry soil weight.
- Preparation: The dry constituents were blended in the correct proportions and mixed thoroughly with a large-scale laboratory mixer.

OTHER TEST CONDITIONS
- Photoperiod: the vessels were covered were plastic films and kept under continuous illumination
- Light intensity: 400 - 800 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality at day 7 and at day 14.
- Other measurements: pH and moisture content at day 0 and at day 14.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: not applicable since the test was performed as a limit test.
- Range finding study test concentrations: 100, 500 and 1000 mg/kg dw
- Results used to determine the conditions for the definitive study: yes, based on the results of the range finding study, a limit test was carried out using a nominal concentration of 1000 mg/kg (dry weight of soil)
Nominal and measured concentrations:
Nominal concentration: 1000 mg/kg dry soil
Reference substance (positive control):
yes
Remarks:
2-Chloracetamide (performed three times a year; most recent test presented in report)
Key result
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
Details on results:
- Mortality at end of exposure period: no mortality occured at the blank control group and at the limit concentration.
- Average weight at the end of exposure: 318 - 353 mg
- Behavioural effects: at the beginning of the administration, test worms soon disappeared into the test soil medium. During the test, all earthworms remained in the soil.
- No unusual circumstances that might have affected the integrity and quality of the study were observed.
Results with reference substance (positive control):
- Results with reference substance valid? Yes
- Relevant effect levels: 14 d LC50: 21.2 mg/kg dry soil, 95% confidence limit 16.9 - 26.6 mg/kg dry soil.
Reported statistics and error estimates:
none

Table 1: Number of Dead Earthworms in the Limit Test

Concentration applied to soil (mg/kg (dry soil))

Number of the test worms

Number the dead

7 d

14 d

0

10

0

0

0

10

0

0

0

10

0

0

0

10

0

0

1000

10

0

0

1000

10

0

0

1000

10

0

0

1000

10

0

0

LC50 (mg/kg (dry soil))

>1000

>1000

Validity criteria fulfilled:
yes
Remarks:
No mortality occured in the control groups.
Conclusions:
The LC50 for acute toxicity to earthworms (14-day exposure) was > 1000 mg/kg (dry soil). The highest concentration causing no mortality at 14 days was 1000 mg/kg (dry soil)
Executive summary:

In an acute toxicity test, performed according to "Guideline HJ/T 153 -2004" and "Guidelines for the Testing of Chemicals-Effects on Biotic Systems", with reference to OECD guideline 207, and according to GLP principles, Eisenia fetida (earthworm) were exposed to a limit concentration (1000 mg/kg (dry soil)) of the test substance for 14 days. A blank control group was included. The treatment group and the control groups consisted of 4 replicates of 10 worms each. The test was performed in artificial soil consisting of sphagnum peat (10%), kaolin clay (20%) and quartz sand (70%). Based on the results of a range finding study in which no mortality occured in all tested concentrations (10, 500 and 1000 mg/kg dry soil), a nominal concentration of 1000 mg/kg dry soil was used in the limit test. Mortality was recorded at day 7 and at day 14. No mortality occured in the blank control group and in the treatment group. The LC50 was determined to be >1000 mg/kg (dry soil), based on the nominal test concentration. The test is considered reliable without restriction.

Description of key information

In an acute toxicity test, performed according to "Guideline HJ/T 153 -2004" and "Guidelines for the Testing of Chemicals-Effects on Biotic Systems", with reference to OECD guideline 207, and according to GLP principles, Eisenia fetida (earthworm) were exposed to a limit concentration (1000 mg/kg (dry soil)) of the test substance for 14 days. A blank control group was included. The treatment group and the control groups consisted of 4 replicates of 10 worms each. The test was performed in artificial soil consisting of sphagnum peat (10%), kaolin clay (20%) and quartz sand (70%). Based on the results of a range finding study in which no mortality occurred in all tested concentrations (10, 500 and 1000 mg/kg dry soil), a nominal concentration of 1000 mg/kg dry soil was used in the limit test. Mortality was recorded at day 7 and at day 14. No mortality occurred in the blank control group and in the treatment group. The LC50 was determined to be >1000 mg/kg (dry soil), based on the nominal test concentration. The test is considered reliable without restriction.

Key value for chemical safety assessment

Additional information

14d-LC50 > 1000 mg/kg, no acute hazard identified.

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