Registration Dossier

Diss Factsheets

Ecotoxicological information

Toxicity to soil macroorganisms except arthropods

Currently viewing:

Administrative data

Link to relevant study record(s)

Referenceopen allclose all

Endpoint:
toxicity to soil macroorganisms except arthropods: long-term
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2021-04-30 to 2021-09-10
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))
Version / remarks:
July 2016
Deviations:
yes
Remarks:
Food was provided on day 0 (test start) instead of day 1 due to good experience with this procedure. Soil moisture: see "Any other information on materials and methods incl. tables", pH-Values, Moisture Room Temperature: see Test conditions, Temperature
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
Test item LICOCARE RBW 300 FL TP
Batch number DEF2114527
Content (certified) 99.75% w/w
CAS No. Not definable
Appearance yellow solid flakes
Water solubility < 1g/L (OECD 105)
Stability under Not specified
test conditions
Expiry date 2022-11-05 (Minimum durability)
Recommended storage Room temperature, in tightly closed container in cool
and well ventilated area, keep away from a source of
ignition

Vehicle:
yes
Remarks:
Quartz sand
Details on preparation and application of test substrate:
The test item was treated with a mortar.
The respective test item amount for each test item concentration was weighed out and mixed with quartz sand (10 g per replicate).
The spiked quartz sand (40 g) was given to the artificial soil (2360 g sdw). Demineralised water (308 g) was added to the artificial soil to adjust the humidity of the artificial soil to a moisture of 54 % of the WHCmax.
Subsequently, the test medium was thoroughly mixed to ensure a homogenous distribution and about 600 g SDW were filled into each test vessel.
Test organisms (species):
Eisenia fetida
Animal group:
annelids
Details on test organisms:
Test system
Eisenia fetida (Annelida, Lumbricidae)

Reason for the selection
Eisenia fetida is suitable for this kind of study and is one of the 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.

Synchronisation of the earthworm population
Earthworms of homogeneous age and body weight were used in
the definitive test. Synchronisation of the population was achieved by placing adult earthworms into breeding boxes and removing the adults after 4 weeks. Offspring from the remaining cocoons reached the adult age after 2 months at the earliest.

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

Number and age of the earthworms
10 adult earthworms (with clitellum) aged 2-12 months were used
per replicate. The difference of age did not deviate by more than 1 month.

Body weight (actual)
0.40 to 0.54 g


Adaption
Earthworms were adapted to the artificial soil to be used for the test two days prior to test start.

Homogeneity of groups
The worms were washed with demineralised water and dried gently on a paper towel. The homogeneity of the population was checked by weighing the earthworms. After confirmation of homogeneity, groups of 10 earthworms (chosen by a randomised procedure) were weighed and introduced into each test vessel.


Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
no
Total exposure duration:
56 d
Test temperature:
Temperature 20 +- 2 °C

The room temperature temporarily deviated from the nominal room temperature for > 8 h in total about 216 hours which is approx. 16 % of the overall test duration (see "Any other information on materials and methods incl. tables",Environmental Conditions). However, its impact on the soil temperature is judged to be negligible as shown by the fulfilment of all validity criteria. This deviation is considered to have no impact on quality and integrity of the study.
pH:
The initial pH-value of the artificial soil was 5.61 (measured on day -7 before the start of exposure).
For the pH-values at test start and end please refer to "Any other information on materials and methods incl. tables",pH-Values, Moisture and WHCmax of the Test Media.
Moisture:
Two days before test start the artificial soil (20000 g) was pre-moistened with 2334 g demineralised water to achieve a moisture content corresponding to 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.

Throughout the test the replicates were weighed weekly to determine water loss via evaporation. Evaporated water was replenished by adding demineralised water to maintain the soil moisture which should not deviate by more than 10% of the initial value at the end of the test. Additional moistening was performed in case the soil appeared too dry.
For the moisture content of the test medium at test start and test end, please refer to "Any other information on materials and methods incl. tables", pH-Values, Moisture and WHCmax of the Test Media).
Details on test conditions:
Test duration
8 weeks

Number of replicates
8 replicates per control and 4 replicates per test item concentration

Test medium
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 % particle size of
0.05 - 0.2 mm)
- 0.35 % calcium carbonate (CaCO3) to achieve a pH of 6.0 +- 0.5

The WHCmax, the moisture content and the pH-value of the artificial soil were determined and adjusted. Two days before test start the artificial soil (20000 g) was pre-moistened with 2334 g demineralised water to achieve a moisture content corresponding to 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 container
Test units containing about 600 g SDW were round plastic boxes with an inner diameter of 15.0 cm corresponding to a bottom surface area of 177 cm2 and a height of 14 cm. Transparent and perforated lids enable sufficient gas exchange, light input and prevent the test substrate from drying.

Temperature 20 +- 2 °C
Photoperiod 16 h light and 8 h dark
Light intensity 400 - 800 lx


Feeding
The earthworms were fed with air-dried cattle manure weekly during the first 4 weeks (see "Any other information on materials and methods incl. tables", Feeding and Additional Moistening During the Test).
At test start and at day 28 the food was mixed carefully into the soil, on the other days the food was placed onto the soil surface. No further feeding took place during the remaining 4 weeks of the test.
The cattle manure was delivered by LEHR- UND FORSCHUNGSGUT RUTHE, Schäferberg 1, 31157 Sarstedt, Germany.
Manure was obtained from cattle that were not subject to medication or treatment with substances, such as growth promoters, nematicides or similar veterinary products that could adversely affect the worms during the test. The manure was air-dried and finely ground before use.

Moistening
Throughout the test the replicates were weighed weekly to determine water loss via evaporation. Evaporated water was replenished by adding demineralised water to maintain the soil moisture which should not deviate by more than 10% of the initial value at the end of the test. Additional moistening was performed in case the soil appeared too dry (see "Any other information on materials and methods incl. tables", Feeding and Additional Moistening During the Test and pH-Values. Moisture and WHCmax of the Test Media).



Nominal and measured concentrations:
Five test item concentrations were tested:

62.5 - 125 - 250 - 500 - 1000 mg product/kg soil dry weight
(factor 2)
The application rates are determined based on the results of an
acute earthworm toxicity test with the same test item (Winkelmann, G., 2018, Study ID: 180706CM / RRA18315).
Reference substance (positive control):
yes
Remarks:
CARBENDAZIM
Key result
Duration:
57 d
Dose descriptor:
LOEC
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Key result
Duration:
57 d
Dose descriptor:
NOEC
Effect conc.:
>= 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Key result
Duration:
28 d
Dose descriptor:
LOEC
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Key result
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
>= 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Key result
Duration:
28 d
Dose descriptor:
LOEC
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Key result
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
>= 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
-No earthworm mortality was observed in the control and all test item concentrations.
-No pathological symptoms or changes in the behaviour of adult earthworms were observed in the control and all test item concentrations.
-At test start, the earthworms had individual weights of 0.40 - 0.54 g. There were no statistically significant differences in biomass in all treatment rates compared to the control.
-The reproduction rate (average number of juveniles produced) was 155 in the control and ranged from 141 to 159 in the test item treatment rates. There were no statistically significant differences in earthworm reproduction at all treatment rates.
The coefficient of variation calculated for the reproduction of the control was 11.0% and thus lower than 30 %.
Results with reference substance (positive control):
The test is conducted once within twelve months. The most recent test was carried out from 2021-02-19 to 2021-04-15 using artificial soil containing 10 % peat (Study ID 72102IRS / RBN70070).
According to OECD 222 significant effects of Carbendazim should be observed between 1 and 5 mg a.i./kg SDW.
For details please refer to "Any other information on results incl. tables",Data of Reference Item.
Reported statistics and error estimates:
Adult Mortality
The arithmetic mean and NOEC were determined.
Since no mortality occurred no further computations were performed.


Biomass
- Shapiro-Wilk’s Test on Normal Distribution
- Levene’s Test on Variance Homogeneity (with Residuals)
- Trend Analysis on Contrasts (Monotonicity of Concentration/Response)
- Dunnett’s Multiple Sequential t-test Procedure
The alpha-value (acceptable probability of incorrectly concluding that there is a difference) was alpha = 0.05. For the endpoint of biomass, the arithmetic mean and the variance (coefficient of variation) per treatment and control were calculated.


Reproduction
- Shapiro-Wilk’s Test on Normal Distribution
- Levene’s Test on Variance Homogeneity (with Residuals)
- Trend Analysis on Contrasts (Monotonicity of Concentration/Response)
- Dunnett’s Multiple t-test Procedure

The alpha-value (acceptable probability of incorrectly concluding that there is a difference) was alpha = 0.05. For the endpoint of reproduction, the arithmetic mean and the variance (coefficient of variation) per treatment and control were calculated. Since the slope of the relationship was found to be not significant, no EC and confidence limits are provided.

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






























































































Application rateReplicate 
[mg test item/kg  SDW]12345678Mean
Control000000000
62.50000    0
1250000    0
2500000    0
5000000    0
10000000    0

 


Pathological Symptoms and Changes in Behaviour





































































































Application rate    Replicate    
[mg test item/kg  SDW]Observation parameter12345678
ControlA10/1010/1010/1010/1010/1010/1010/1010/10
62.5A10/1010/1010/1010/10    
125A10/1010/1010/1010/10    
250A10/1010/1010/1010/10    
500A10/1010/1010/1010/10    
1000A10/1010/1010/1010/10    

A = no obvious pathological symptoms


 


Body Weight Changes of the Adult Earthworms























































































































































































































































































































































Application rateReplicateMean body weights per replicateMean body weight change of earthworms   
[mg test item/kg  SDW] Test start28 daysPer replicateMean ± SD Sig.
  [g][g][g][%][g][%] 
Control10.470.470.000.00.03 ± 0.025.55 ± 5.02
 20.480.47-0.01-2.1   
 30.470.510.048.5   
 40.480.540.0612.5   
 50.480.490.012.1   
 60.470.500.036.4   
 70.470.510.048.5   
 80.470.510.048.5   
62.510.480.46-0.02-4.20.02 ± 0.034.25 ± 6.69No
 20.480.490.012.1   
 30.470.510.048.5   
 40.470.520.0510.6   
12510.470.500.036.40.04 ± 0.028.40 ± 3.72No
 20.480.530.0510.4   
 30.480.540.0612.5   
 40.470.490.024.3   
25010.480.550.0714.60.07 ± 0.0214.2 ± 4.45No
 20.470.540.0714.9   
 30.470.560.0919.1   
 40.480.520.048.3   
50010.470.520.0510.60.04 ± 0.027.38 ± 5.03No
 20.480.480.000.0   
 30.480.520.048.3   
 40.470.520.0510.6   
100010.470.530.0612.80.03 ± 0.026.90 ± 4.07No
 20.480.500.024.2   
 30.480.500.024.2   
 40.470.500.036.4   

SD = Standard deviation
Sig. = Statistical Significance (Dunnett’s Multiple Sequential t-test Procedure, alpha = 0.05) compared to control


 


 


Reproduction Rate (Number of Juveniles after 8 Weeks)



















































































































































































































































































Application rateReplicateNumber of juvenilesMean ± SDCV               [%][%]Sig.
[mg test item/kg SDW]    of control 
Control1182155 ± 17.111.0--
 2181    
 3145    
 4140    
 5152    
 6147    
 7141    
 8148    
62.51135150 ± 27.918.696.8No
 2132    
 3191    
 4140    
1251191159 ± 26.116.4103No
 2135    
 3141    
 4170    
2501135141 ± 17.712.591.0No
 2165    
 3142    
 4123    
5001137154 ± 14.89.5899.4No
 2152    
 3173    
 4  154*    
10001134150 ± 23.115.496.8No
 2134    
 3183    
 4149    

SD = Standard deviation CV = Coefficient of variation
Sig. = Statistical Significance (Dunnett’s Multiple t-test Procedure, α = 0.05) compared to the control
*) One adult worm was found on day 56, not taken into consideration


 

Validity criteria fulfilled:
yes
Conclusions:
Under the conditions of this study, neither earthworm mortalities nor pathological symptoms or changes in the behaviour of adult earthworms were observed in the control or all test item concentrations. There were no statistically significant differences in biomass in all test item concentrations compared to the control. After a further four weeks, the reproduction rate (average number of juveniles produced) was 155 in the control and ranged from 141 to 159 in the test item treatment rates. There were no statistically significant differences in earthworm reproduction in all treatment rates compared to the control. No effect values could be determined for biomass, reproduction and mortality, as no effects occurred up to 1000 mg test item/kg SDW.
All validity criteria recommended by the test guidelines were fulfilled.
Executive summary:

Effects of LICOCARE RBW 300 FL TP (batch number: DEF2114527) on mortality, biomass and the reproductive potential of the earthworm species Eisenia fetida (Annelida, Lumbricidae) were determined according to OECD 222 (2016) from 2021-04-30 to 2021-09-10 with the definitive exposure phase from 2021-07-15 to 2021-09-09 at the test facility in 31157 Sarstedt, Germany.


The study was conducted under static conditions over 8 weeks with the test item concentrations 62.5 – 125 – 250 – 500 – 1000 mg LICOCARE RBW 300 FL TP/kg SDW. Each application rate was mixed with an appropriate amount of quartz sand followed by mixing into artificial soil containing 10 % peat. A control including untreated artificial soil was tested under the same conditions as the test item treatments. 80 test organisms were inserted into 8 control replicates. 40 test organisms were divided into 4 replicates for each test item treatment (10 earthworms per replicate). They had an individual body weight between 0.40 and 0.54 g at experimental starting.


After 28 days of exposure in soil, neither earthworm mortalities nor pathological symptoms or changes in the behaviour of adult earthworms were observed in the control or all test item concentrations. There were no statistically significant differences in biomass in all test item concentrations compared to the control. After a further four weeks, the reproduction rate (average number of juveniles produced) was 155 in the control and ranged from 141 to 159 in the test item treatment rates. There were no statistically significant differences in earthworm reproduction in all treatment rates compared to the control. No effect values could be determined for biomass, reproduction and mortality, as no effects occurred up to 1000 mg test item/kg SDW.


All validity criteria recommended by the test guidelines were fulfilled.


 


Summary of All Observed Effects in the Application rates































































EffectsLICOCARE RBW 300 FL TP
Application ratesControl62.51252505001000
[mg product/kg  SDW]
Mean Mortality of adult earthworms [%]000000
Biomass [mean body-weight change* after 28 days of exposure in %] 5.64.38.4147.46.9
Reproduction-96.810391.099.496.8
[% of control]
Summary of all effects-NoNoNoNoNo
[Statistical significance compared to the control]

*) increase of body weight


 


 


NOEC, LOEC and EC-values

















Endpoint[mg test item/kg  SDW]
LOECmortality, biomass, reproduction> 1000
NOECmortality, biomass, reproduction≥ 1000

 

Endpoint:
toxicity to soil macroorganisms except arthropods: short-term
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2018-11-14 to 2018-11-29 with the definitive exposure phase from 2018-11-14 to 2018-11-28
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 207 (Earthworm, Acute Toxicity Tests)
Version / remarks:
1984
Deviations:
yes
Remarks:
6 of 12 in total used glass vessels had the wrong volume (2 L instead of 1.5 L). This deviation is considered to have no impact on quality and integrity of the study.
GLP compliance:
yes
Analytical monitoring:
no
Vehicle:
yes
Remarks:
Quartz sand
Details on preparation and application of test substrate:
The test item was treated with a mortar.
The respective test item amount was weighed out (nominal weight: 3330 mg; actual weight: 3330.3 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 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.
Test organisms (species):
Eisenia fetida
Animal group:
annelids
Details on test organisms:
TEST ORGANISM
- Common name: Eisenia fetida (Annelida, Lumbricidae)
- Source: Breeding stock culture maintained at the test facility
- Age at test initiation (mean and range, SD): Adult
- Weight at test initiation (mean and range, SD): 0.41 +/- 0.03

ACCLIMATION
- Acclimation period: 2 days
- Acclimation conditions (same as test or not): same as test
- Health during acclimation (any mortality observed): no mortality observed
Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
yes
Total exposure duration:
14 d
Test temperature:
17.5 - 22 °C
pH:
Test item concentration
[mg/kg SDW]
pH-value

day 0 day 14
Control 5.99 5.64
1000 5.97 5.60
Moisture:
Test item concentration
[mg/kg SDW]

Moisture [%] of DW
day 0 day 14
Control 26.0 24.9
1000 26.1 24.8
Details on test conditions:
TEST SYSTEM
- Test containers Glass vessels (volume 1.5 L and 2 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.
- Amount of soil or substrate: 555 g soil dry weight
- No. of organisms per container (treatment): 10 per replicate
- No. of replicates per treatment group: 6
- No. of replicates per control: 6

Artificial soil:
- 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)
- 0.38 % 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 (7840 g sdw) was pre-moistened with 883 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.



OTHER TEST CONDITIONS
- Photoperiod: 16 h
- Light intensity: 529 +/- 84

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
The live weight of the earthworms was determined individually on the day of application (day 0) and on day 14.
The mean weight change was assessed on day 14 after application.
Mortality, behaviour and morphological changes of the earthworms were recorded 7 and 14 days after application.


VEHICLE CONTROL PERFORMED: no

TEST CONCENTRATIONS
- Spacing factor for test concentrations: none, limit test
- Justification for using less concentrations than requested by guideline:
- Range finding study
- Test concentrations: 1 - 10 - 100 - 1000 mg/kg soil dry weight
- Results used to determine the conditions for the definitive study:
Range Finding Test (14 days): Mortality and Change of Live Weight
Concentration Mortality of adult earthworms Change of Live weight
[mg test item/kg SDW] [%] [%]
Control 0 -17
1 0 -15
10 0 -19
100 0 -14
1000 0 -13
Nominal and measured concentrations:
1000 mg/kg soil dry weight
Reference substance (positive control):
yes
Remarks:
2-Chloroacetamide
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: Mortality, behaviour, pathological symptoms, change of body weight
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:
- Mortality at end of exposure period: No mortality was observed in the control and the limit test item concentration 1000 mg/kg SDW after 14 days of exposure
- Total mass of adults at beginning of test: Control: 24.7 g; 1000 mg/kg soil dry weight: 24.8 g
- Changes in body weight of live adults (% of initial weight) at end of exposure period: 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.
- Morphological abnormalities: no pathological symptoms or changes in earthworm behaviour were observed in the control and the limit test item concentration after 14 days of exposure

Results with reference substance (positive control):
- Results with reference substance valid? yes
- Relevant effect levels:
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 42.3 n.d.*
*) not determined due to mathematical reasons (3 reference item concentrations)

Reported statistics and error estimates:
NOEC, LOEC and statistical Since no mortality was observed, no further computation was
analyses carried out.
The NOEC/LOEC for biomass was determined using a Two-sample t-test procedure for determination of statistically significant differences in biomass compared to the control. When running the test, a Shapiro-Wilk’s Test on Normal Distribution and Brown-Forsythe test on Equal Variance were done first. The alpha-value (acceptable probability of incorrectly concluding that there is a difference) is alpha = 0.05.

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

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

Mean Live Weight and Mean Loss of Body Weight

 

Mean live weight

Test item

 concentration 

[mg/kg SDW]

Test start

MV±SD

[g/worm]

Test end

MV±SD

[g/worm]

Inhibition

[%]*

Statistically

significant vs.

 Control

Control

0.41 ± 0.03

0.43 ± 0.05

-5

1000

0.41 ± 0.03

0.42 ± 0.04

-2

No

MV = Mean value           SD = Standard deviation     

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 300 on the earthworm Eisenia fetida were determined according to the OECD Guideline 207 at the test facility in 31157 Sarstedt, Germany from 2018-11-14 to 2018-11-29 with thedefinitive exposure phase from 2018-11-14 to 2018-11-28. 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 0.34 - 0.49 g, divided into six replicates, were tested per limit test item concentration and control.

No mortality and no pathological symptoms or changes in earthworm behaviour were observed in the control and thelimit test item concentration after 14 days ofexposure. 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 .

 

All validity criteria of the test guideline were fulfilled.

 

Summary of all Effects

Effects

Licocare RBW 300

[mg/kg SDW]

LOEC

Mortality, behaviour, pathological symptoms,               change of body weight

> 1000

NOEC

Mortality, behaviour, pathological symptoms,             change of body weight

1000


Description of key information

OECD 207


The acute effects of Licocare RBW 300 on the earthwormEisenia fetidawere determined according to the OECD Guideline 207


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.


 


OECD 222


Under the conditions of this study, neither earthworm mortalities nor pathological symptoms or changes in the behaviour of adult earthworms were observed in the control or all test item concentrations. There were no statistically significant differences in biomass in all test item concentrations compared to the control. After a further four weeks, the reproduction rate (average number of juveniles produced) was 155 in the control and ranged from 141 to 159 in the test item treatment rates. There were no statistically significant differences in earthworm reproduction in all treatment rates compared to the control. No effect values could be determined for biomass, reproduction and mortality, as no effects occurred up to 1000 mg test item/kg SDW.
All validity criteria recommended by the test guidelines were fulfilled.

Key value for chemical safety assessment

Long-term EC10, LC10 or NOEC for soil macroorganisms:
1 000 mg/kg soil dw

Additional information