Polar modified Rice Bran Wax

Toxicity to soil macroorganisms except arthropods

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:

11.2019

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)

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

Test organism Eisenia fetida (Annelida, Lumbricidae)

Reason for the selection Eisenia fetida is suitable for this kind of study and is one of the
of test system recommended species according to the guideline.

Source Breeding stock culture maintained at the test facility

Breeding Organisms of the species Eisenia fetida are bred at the test facility in covered plastic vessels containing potting compost.

Breeding conditions Temperature of 20 ± 5°C in the dark

Feeding During breeding, the earthworms are fed with litter of dried stinging nettle leaves and porridge oats. A sufficient amount of food is provided depending on the feeding rate and the density of the earthworm population in the vessels.

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

yes

Total exposure duration:

14 d

Test temperature:

20 +/- 2 °C

pH:

5.89 - 6.31

Moisture:

23.3 % - 25.1 %

Details on test conditions:

Test duration 2 weeks

Number of replicates 6 replicates were used for the control and the limit test item concentration.

Number and age of 10 adult earthworms (aged 2 – 12 months, with clitellum) with an
the earthworms individual live weight between 0.45 – 0.55 g were used per replicate.

Adaptation The earthworms were conditioned to artificial soil two days prior to the start of exposure.

Test media Artificial soil consisted of the following components:
- 10 % Peat, air-dried and finely ground
- 20 % Kaolin, kaolinite content > 30 %
- 69 % Air-dried quartz sand (sand with > 50 % particles size
of 0.05 to 0.2 mm)
- 0.40 % calcium carbonate (CaCO3) to adjust the pH to 6.0 ±
0.5
The soil moisture content and the WHCmax were determined prior to experimental starting.
Two days before earthworm insertion into the test vessels the artificial soil (9700 g sdw) was pre-moistened with 990 g deionised water to achieve a moisture of 27 % of the WHCmax of the artificial soil. The moistened soil was kept covered to prevent evaporative water losses prior to the start of exposure.



Test containers Glass vessels (volume 1.5 L, used for preserving) were used. During the test, transparent and perforated lids enabled sufficient gas exchange and light input and prevented drying of the test substrate.

Application The test item was treated with a mortar.
The respective test item amount was weighed out (nominal weight: 3330 mg; actual weight: 3330.2 mg) for the limit test item concentration and mixed with quartz sand (10 g per replicate). The spiked quartz sand was given to the artificial soil. Demineralised water (490 g) was added to the artificial soil to adjust the humidity of the artificial soil to a moisture of 54 % of the maximum water holding capacity. Subsequently, the test medium was thoroughly mixed to ensure a homogenous distribution and about 555 g SDW were filled into each test vessel.

Feeding No food was added to the test vessels.

Temperature 20 ± 2°C

Photoperiod 24 h

Light intensity 400 - 800 lx, measured at least every 6 months

TEST SYSTEM
- Test container (material, size):
- Amount of soil or substrate:
- No. of organisms per container (treatment):
- No. of replicates per treatment group:
- No. of replicates per control:
- No. of replicates per vehicle control:

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Geographic location:
- Pesticide use history at the collection site:
- Collection procedures:
- Sampling depth (cm):
- Soil texture (if natural soil)
- % sand:
- % silt:
- % clay:
- Soil taxonomic classification:
- Soil classification system:
- Composition (if artificial substrate):
- Organic carbon (%):
- Maximum water holding capacity (in % dry weigth):
- CEC:
- Pretreatment of soil:
- Storage (condition, duration):
- Stability and homogeneity of test material in the medium:

OTHER TEST CONDITIONS
- Photoperiod:
- Light intensity:

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :

VEHICLE CONTROL PERFORMED: yes/no

TEST CONCENTRATIONS
- Spacing factor for test concentrations:
- Justification for using less concentrations than requested by guideline:
- Range finding study
- Test concentrations:
- Results used to determine the conditions for the definitive study:
TEST SYSTEM
- Test container (material, size):
- Amount of soil or substrate:
- No. of organisms per container (treatment):
- No. of replicates per treatment group:
- No. of replicates per control:
- No. of replicates per vehicle control:

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Geographic location:
- Pesticide use history at the collection site:
- Collection procedures:
- Sampling depth (cm):
- Soil texture (if natural soil)
- % sand:
- % silt:
- % clay:
- Soil taxonomic classification:
- Soil classification system:
- Composition (if artificial substrate):
- Organic carbon (%):
- Maximum water holding capacity (in % dry weigth):
- CEC:
- Pretreatment of soil:
- Storage (condition, duration):
- Stability and homogeneity of test material in the medium:

OTHER TEST CONDITIONS
- Photoperiod:
- Light intensity:

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :

VEHICLE CONTROL PERFORMED: yes/no

TEST CONCENTRATIONS
- Spacing factor for test concentrations:
- Justification for using less concentrations than requested by guideline:
- Range finding study
- Test concentrations:
- Results used to determine the conditions for the definitive study:

Nominal and measured concentrations:

1000 mg/kg SDW

Reference substance (positive control):

yes

Remarks:

Reference item 2-Chloroacetamide (SIGMA-ALDRICH) Test performance A reference test in artificial soil is conducted once every twelve months. For details please refer to part 6.3. Test concentrations 15 – 30 – 60 mg/kg SDW Purpose To determine the sens

Key result

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:

other: Mortality, behaviour, pathological symptoms, change of body weight

Details on results:

No mortality was observed in the control and the limit test item concentration 1000 mg/kg SDW after 14 days of exposure (Table 2).

Table 2: Earthworm Mortality in [%] after 7 and 14 Days of Exposure
Mortality [%]
Test item concentration
[mg/kg SDW] Day 7 Day 14
1 2 3 4 5 6 MV 1 2 3 4 5 6 MV
Control 0 0 0 0 0 0 0 0 0 0 0 0 0 0
1000 0 0 0 0 0 0 0 0 0 0 0 0 0 0
MV = mean value
No pathological symptoms nor changes in the earthworm behaviour were observed in the control as well as in the limit test item concentration 1000 mg/kg SDW after 7 and 14 days of exposure (Table 3).

Table 3: Earthworm Behaviour and Pathological Symptoms after 7 and 14 Days of Exposure
Test item concentration [mg/kg SDW] Effect Replicate
1 2 3 4 5 6
Day 7
Control A) 10/10 10/10 10/10 10/10 10/10 10/10
1000 A) 10/10 10/10 10/10 10/10 10/10 10/10
Day 14
Control A) 10/10 10/10 10/10 10/10 10/10 10/10
1000 A) 10/10 10/10 10/10 10/10 10/10 10/10
A) no obvious pathological symptoms or behavioural changes

Results with reference substance (positive control):

The acute toxicity of 2-Chloroacetamide on the earthworm Eisenia fetida fetida (SAVIGNY) was determined over a period of two weeks from 2019-01-30 to 2019-02-14 (71901IRS/RRA70069). The LC50 value was in the range between 20 and 80 mg/kg SDW as recommended by the guideline (Table 7).

Table 7: NOEC and LC-Value after 14 d for the Reference Item 2-Chloroacetamide
LC-Value /
NOEC Concentration
[mg reference item/kg SDW] 95 % - Confidence Interval
[mg reference item/kg SDW]
NOEC 30 –
LC50 50.5 n.d.*
*) not determined due to mathematical reasons (3 reference item concentrations)

Validity criteria fulfilled:

yes

Conclusions:

No mortality and no pathological symptoms or changes in earthworm behaviour were observed in the control and the limit test item concentration after 14 days of exposure. Therefore, the NOEC was determined to be 1000 mg test item/kg SDW.
In terms of body weight loss, no statistically significant effect was observed in the limit test item concentration after 14 days of exposure compared to the control. Consequently, the NOEC was determined to be 1000 mg test item/kg SDW.

Executive summary:

The acute effects of LICOCARE RBW 106 FL TP  on the earthworm Eisenia fetida were determined according to the OECD Guideline 207 at the test facility in 31157 Sarstedt, Germany from 2019-11-14 to 2019-11-28. The study was conducted under static conditions for 14 days with a limit test item concentration of 1000 mg/kg SDW mixed into artificial soil containing 10 % peat. Artificial soil without test item was used as control. 60 test organisms with an individual weight between 0.45 - 0.55 g, divided into six replicates, were tested per limit test item concentration and control, each.

No mortality and no pathological symptoms or changes in earthworm behaviour were observed in the control and the limit test item concentration after 14 days of exposure. Therefore, the NOEC was determined to be 1000 mg test item/kg SDW (Table 1).

In terms of body weight loss, no statistically significant effect was observed in the limit test item concentration after 14 days of exposure compared to the control. Consequently, the NOEC was determined to be 1000 mg test item/kg SDW (Table 1).

 

All validity criteria of the test guideline were fulfilled.

Description of key information

No mortality and no pathological symptoms or changes in earthworm behaviour were observed in the control and the limit test item concentration after 14 days of exposure. Therefore, the NOEC was determined to be 1000 mg test item/kg SDW.

In terms of body weight loss, no statistically significant effect was observed in the limit test item concentration after 14 days of exposure compared to the control. Consequently, the NOEC was determined to be 1000 mg test item/kg SDW.

Key value for chemical safety assessment

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

1 000 mg/kg soil dw