Registration Dossier
Registration Dossier
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EC number: 911-436-4 | CAS number: 61932-63-6
Toxicity to soil macroorganisms except arthropods
Administrative data
Link to relevant study record(s)
- Endpoint:
- toxicity to soil macroorganisms except arthropods: long-term
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2010-01-08 to 2010-03-09
- Reliability:
- 1 (reliable without restriction)
- Qualifier:
- according to guideline
- 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:
- Room temperature decreased several times to a minimum of 17 °C for a total time of about 11 hours. The soil moisture of all test vessels deviated by more than 10 % of the initial moisture at the end of the test since it was decided to add additional water
- GLP compliance:
- yes
- Analytical monitoring:
- no
- Vehicle:
- yes
- Details on preparation and application of test substrate:
- - Method of mixing into soil (if used): The respective test item concentration was weighed out, dissolved 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. 1164 mL demineralised water were added 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 600 g soil dry weight were filled into each test vessel.
- Controls: Artificial soil with addition of quartz sand moistened with demineralised water without test item.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone, organic solvent
- 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 (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 and range, SD): 0.53 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 type:
- laboratory study
- Substrate type:
- artificial soil
- Limit test:
- yes
- Total exposure duration:
- 56 d
- Remarks:
- Removement of adults after 28 days
- Test temperature:
- 20 +/- 2°C
- pH:
- Application rate
[mg test item/kg soil dry weight] pH-value
day 0 day 56
Control 5.75 6.25
Solvent Control 5.80 6.45
1000 5.86 6.29 - Moisture:
- Application rate
[mg test item/kg soil dry weight] Moisture [%] of dry weight
day 0 day 56
Control 37.9 43.8*
Solvent Control 37.3 44.7*
1000 38.2 42.9* - 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
SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Composition (if artificial substrate): 10 % peat, 20 % kaolin, 69 % air dried quartz sand, 0.39 % calcium carbonate
- Organic carbon (%): not determined
- Maximum water holding capacity (in % dry weigth): 73
- CEC: not determined
- Pretreatment of soil: none
- Storage (condition, duration): 18 days after preparation at test conditions in the dark
OTHER TEST CONDITIONS
- Photoperiod: 16 h light - 8 h dark
- Light intensity: 649 +/- 79 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
TEST CONCENTRATIONS
- Spacing factor for test concentrations: None, limit test concentration - 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
- 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
- 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 body weights of the earthworms increased in the control and at the limit test item concentration of 1000 mg test item/kg soil dry weight throughout the first 28 days. Compared to the control, no statistically significant difference in the increase of biomass was found
- No. of offspring produced: The mean reproduction rate was 332 in the pooled control and 375 at the limit test item concentration of 1000 mg test item/kg soil dry weight. Compared to the pooled control, the reproduction rate was not statistically significantly reduced, but increased at 1000 mg test item/kg soil dry weight. The coefficient of variation calculated for the reproduction of both controls was lower than 30 %
- 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 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 ¿-value for ANOVA test (acceptable probability of incorrectly concluding that there is a difference) was ¿ = 0.05. Due to a failed Normality Test reproduction data were log-transformed prior to running the ANOVA.
- Validity criteria fulfilled:
- yes
- Conclusions:
- Under the conditions of this study, Pigment Red 112 did not induce mortality or pathological symptoms of adult earthworms after an exposure to the test item concentration of 1000 mg test item/kg soil dry weight for 28 days. 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 was not statistically significantly reduced, but increased 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/ kg soil dry weight. - Executive summary:
Effects of Pigment Red 112 on mortality, biomass and the reproductive potential of the earthworm species Eisenia 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 a limit test item concentration of 1000 mg test item/kg soil dry weight, which was mixed into artificial soil containing 10 % peat. A control using untreated artificial soil and a solvent control with the solvent mixed into the artificial soil were set up. 80 test organisms, with an individual weight of 0.30 to 0.60 g, were divided into 8 replicate groups with 10 test organisms per replicate.
After 28 days no evident mortality as well as no pathological symptoms or changes in the behaviour of adult earthworms were seen in the controls as well as at the limit test item concentration of 1000 mg test item/kg soil dry weight. The biomass of the adult earthworms increased in the pooled control and at the limit test item concentration without a statistically significant difference. After further four weeks the reproduction rate (average number of juveniles) was 332 in the pooled control and 375 at the limit test item concentration. Compared to the pooled control, the reproduction rate was not statistically significantly reduced.
Overall, the NOEC of the test item concerning mortality, biomass and reproduction of earthworms was determined to be at 1000 mg test item/kg soil dry weight.
All validity criteria recommended by the test guidelines were fulfilled.
Summary of All Observed Effects in the Tested Dose
Effects
Test item
[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
- Endpoint:
- toxicity to soil macroorganisms except arthropods: long-term
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Study period:
- 2010-01-08 to 2010-03-09
- Reliability:
- 1 (reliable without restriction)
- Justification for type of information:
- See Rationale and Justification for the Analogue Read-Across Approach for the registration of the Nanoform of Pigment Red 210 (Chapter 13)
- Reason / purpose for cross-reference:
- read-across source
- Qualifier:
- according to guideline
- 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:
- Room temperature decreased several times to a minimum of 17 °C for a total time of about 11 hours. The soil moisture of all test vessels deviated by more than 10 % of the initial moisture at the end of the test since it was decided to add additional water
- GLP compliance:
- yes
- Analytical monitoring:
- no
- Vehicle:
- yes
- Details on preparation and application of test substrate:
- - Method of mixing into soil (if used): The respective test item concentration was weighed out, dissolved 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. 1164 mL demineralised water were added 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 600 g soil dry weight were filled into each test vessel.
- Controls: Artificial soil with addition of quartz sand moistened with demineralised water without test item.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone, organic solvent
- 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 (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 and range, SD): 0.53 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 type:
- laboratory study
- Substrate type:
- artificial soil
- Limit test:
- yes
- Total exposure duration:
- 56 d
- Remarks:
- Removement of adults after 28 days
- Test temperature:
- 20 +/- 2°C
- pH:
- Application rate
[mg test item/kg soil dry weight] pH-value
day 0 day 56
Control 5.75 6.25
Solvent Control 5.80 6.45
1000 5.86 6.29 - Moisture:
- Application rate
[mg test item/kg soil dry weight] Moisture [%] of dry weight
day 0 day 56
Control 37.9 43.8*
Solvent Control 37.3 44.7*
1000 38.2 42.9* - 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
SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Composition (if artificial substrate): 10 % peat, 20 % kaolin, 69 % air dried quartz sand, 0.39 % calcium carbonate
- Organic carbon (%): not determined
- Maximum water holding capacity (in % dry weigth): 73
- CEC: not determined
- Pretreatment of soil: none
- Storage (condition, duration): 18 days after preparation at test conditions in the dark
OTHER TEST CONDITIONS
- Photoperiod: 16 h light - 8 h dark
- Light intensity: 649 +/- 79 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
TEST CONCENTRATIONS
- Spacing factor for test concentrations: None, limit test concentration - 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
- 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
- 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 body weights of the earthworms increased in the control and at the limit test item concentration of 1000 mg test item/kg soil dry weight throughout the first 28 days. Compared to the control, no statistically significant difference in the increase of biomass was found
- No. of offspring produced: The mean reproduction rate was 332 in the pooled control and 375 at the limit test item concentration of 1000 mg test item/kg soil dry weight. Compared to the pooled control, the reproduction rate was not statistically significantly reduced, but increased at 1000 mg test item/kg soil dry weight. The coefficient of variation calculated for the reproduction of both controls was lower than 30 %
- 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 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 ¿-value for ANOVA test (acceptable probability of incorrectly concluding that there is a difference) was ¿ = 0.05. Due to a failed Normality Test reproduction data were log-transformed prior to running the ANOVA.
- Validity criteria fulfilled:
- yes
- Conclusions:
- Under the conditions of this study, Pigment Red 112 did not induce mortality or pathological symptoms of adult earthworms after an exposure to the test item concentration of 1000 mg test item/kg soil dry weight for 28 days. 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 was not statistically significantly reduced, but increased 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/ kg soil dry weight. - Executive summary:
Effects of Pigment Red 112 on mortality, biomass and the reproductive potential of the earthworm species Eisenia 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 a limit test item concentration of 1000 mg test item/kg soil dry weight, which was mixed into artificial soil containing 10 % peat. A control using untreated artificial soil and a solvent control with the solvent mixed into the artificial soil were set up. 80 test organisms, with an individual weight of 0.30 to 0.60 g, were divided into 8 replicate groups with 10 test organisms per replicate.
After 28 days no evident mortality as well as no pathological symptoms or changes in the behaviour of adult earthworms were seen in the controls as well as at the limit test item concentration of 1000 mg test item/kg soil dry weight. The biomass of the adult earthworms increased in the pooled control and at the limit test item concentration without a statistically significant difference. After further four weeks the reproduction rate (average number of juveniles) was 332 in the pooled control and 375 at the limit test item concentration. Compared to the pooled control, the reproduction rate was not statistically significantly reduced.
Overall, the NOEC of the test item concerning mortality, biomass and reproduction of earthworms was determined to be at 1000 mg test item/kg soil dry weight.
All validity criteria recommended by the test guidelines were fulfilled.
Summary of All Observed Effects in the Tested Dose
Effects
Test item
[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
- Endpoint:
- toxicity to soil macroorganisms except arthropods: long-term
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2009-06-10 to 2009-08-06
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: Guideline study compliant to GLP
- Qualifier:
- according to guideline
- 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:
- Room temperature increased once to about 22.5 °C for four hours. The soil moisture of all test vessels deviated by more than 10 % of the initial value at the end of the test.
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- GLP according to German Chemikaliengesetz and Directive 88/320/EEC
- Analytical monitoring:
- no
- Vehicle:
- yes
- Details on preparation and application of test substrate:
- - Method of mixing into soil (if used): The respective test item concentration was weighed out, dissolved 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 52 % of the maximum water holding capacity. Subsequently, the test medium will be mixed with a mixer for at least three minutes and 600 g soil dry weight were filled into each test vessel.
- Controls: Artificial soil with addition of quartz sand moistened with demineralised water without test item.
- 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 acetone per replicate, 10 g quartz sand per replicate
- Evaporation of vehicle before use: Yes - 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 and range, SD): 0.44 +/- 0.05 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 type:
- laboratory study
- Substrate type:
- artificial soil
- Limit test:
- no
- Total exposure duration:
- 56 d
- Remarks:
- Removement of adults after 28 days
- Test temperature:
- 18 - 22.5 deg C
- pH:
- Application rate
[mg test item/kg soil DW] pH-value
day 0 day 56
Control 5.42 6.13
Solv. Control 5.40 6.40
62.5 5.39 6.18
125 5.39 6.30
250 5.40 6.33
500 5.38 6.07
1000 5.42 6.14 - Moisture:
- Application rate
[mg test item/kg soil DW] Moisture [%] of dry weight
day 0 day 56
Control 35.6 42.5*
Solv. Control 35.1 45.8*
62.5 34.8 46.7*
125 35.5 46.8*
250 34.9 44.6*
500 35.2 45.2*
1000 34.6 44.6*
*Variation of the water content > 10 %, this deviation is considered to have no impact on the study - 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: 4
- No. of replicates per control: 4
- No. of replicates per vehicle control: 4
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): 68
- CEC: not determined
- Pretreatment of soil: none
- Storage (condition, duration): 8 days after preparation at test conditions in the dark
OTHER TEST CONDITIONS
- Photoperiod: 16 h light - 8 h dark
- Light intensity: 524 +/- 70 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
TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2 - Nominal and measured concentrations:
- Nominal application rates: 62.5 - 125 - 250 - 500 - 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
- 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 mortality of adult earthworms was observed in the controls as well as at all application rates after 28 days of exposure to the test item
- 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 body weights of the earthworms increased in the control and at the test item concentrations of 62.5, 125, 250, 500 and 1000 mg test item / kg soil dry weight throughout the first 28 days. The biomass did not differ statistically significant compared to the pooled control at all test item concentrations
- No. of offspring produced: The mean number of juveniles in the pooled control was 359. The average number of juveniles in the treated groups ranged between 288 and 365. Compared to the pooled control the reproduction of the earthworms did not statistically significantly differ at all test item application rates
- Behavioural abnormalities: No symptoms or changes in the behaviour of the earthworms were observed in the controls as well as at all application rates after 28 days of exposure to the test item - 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:
- Differences between the control and the solvent control were statistically analysed using t-tests. No statistical differences occurred between the controls concerning biomass and reproduction. Thus, both controls were pooled for further statistical analyses.
One Way Analysis of Variance (ANOVA) was carried out for the determination of statistically significant differences compared to
the control. A Normality Test and an Equal Variance Test were conducted prior to running the ANOVA. P-values for both Normality and Equal Variance Test are 0.05. The a-value for ANOVA test (acceptable probability of incorrectly concluding that there is a difference) is a = 0.05. - Validity criteria fulfilled:
- yes
- Conclusions:
- In this earthworm reproduction test according to OECD 222 the submission substance did not induce mortality or pathological symptoms of adult earthworms after an exposure to the test item concentrations of 62.5, 125, 250, 500 and 1000 mg/kg soil dry weight for 28 days. Also, the biomass increase of the adult earthworms after 28 days did not statistically significantly differ compared to the pooled control at all test item concentrations. In addition, after eight weeks of exposure the earthworm reproduction was not statistically significantly reduced at all applied test item concentrations.
Thus the No Observed Effect Concentration (NOEC) for the submission substance with respect to mortality, body weight and reproduction of Eisenia fetida fetida after 8 weeks of exposure in artificial soil was determined to be 1000 mg test item/kg soil dry weight. - Executive summary:
Effects of the submission substance on mortality, biomass and the reproductive potential of the earthworm species Eisenia fetida (Savigny) were determined compliant to GLP according to the guidelines DIN ISO 11268-2 (1998), OECD 222 (2004) and BBA (1994) (reliability category 1). The study was conducted under static conditions over 8 weeks with five different application rates (62.5 - 125 - 250 - 500 - 1000 mg test item/kg soil dry weight), which were mixed into artificial soil containing 10 % peat. 40 test organisms, with an individual weight of 0.30 to 0.60 g, were divided into 4 replicate groups with 10 test organisms per replicate.
After 28 days no mortality as well as no pathological symptoms or changes in the behaviour of adult earthworms were seen in the control, in the solvent control and at all tested application rates. The earthworm biomass increased in the control and at all application rates over 28 days without any statistically significant differences compared to the control. After further four weeks the reproduction rate (average number of juveniles) was 359 in the control (control and solvent control) and ranged between 288 and 365 at the test item application rates. Compared to the control, the reproduction rate did not statistically significantly differ at all application rates.
Overall, the NOEC of the submission substance concerning mortality, biomass and reproduction of earthworms was determined to be >= 1000 mg test item/kg soil dry weight (Table 2).
All validity criteria recommended by the test guidelines were fulfilled.
Summary of All Observed Effects in the Tested Doses
Effects
Test item
mg test item/kg soil dry weight
62.5
125
250
500
1000
Mortality of adult earthworms
No
No
No
No
No
Reduction of body weight
No
No
No
No
No
Reduced reproduction rate
(Number of juveniles)
No
No
No
No
No
Summary of all effects
No
No
No
No
No
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
- Endpoint:
- toxicity to soil macroorganisms except arthropods: long-term
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Study period:
- 2009-06-10 to 2009-08-06
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: Guideline study compliant to GLP
- Justification for type of information:
- See Rationale and Justification for the Analogue Read-Across Approach for the registration of the Nanoform of Pigment Red 210 (Chapter 13)
- Reason / purpose for cross-reference:
- read-across source
- Qualifier:
- according to guideline
- 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:
- Room temperature increased once to about 22.5 °C for four hours. The soil moisture of all test vessels deviated by more than 10 % of the initial value at the end of the test.
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- GLP according to German Chemikaliengesetz and Directive 88/320/EEC
- Analytical monitoring:
- no
- Vehicle:
- yes
- Details on preparation and application of test substrate:
- - Method of mixing into soil (if used): The respective test item concentration was weighed out, dissolved 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 52 % of the maximum water holding capacity. Subsequently, the test medium will be mixed with a mixer for at least three minutes and 600 g soil dry weight were filled into each test vessel.
- Controls: Artificial soil with addition of quartz sand moistened with demineralised water without test item.
- 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 acetone per replicate, 10 g quartz sand per replicate
- Evaporation of vehicle before use: Yes - 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 and range, SD): 0.44 +/- 0.05 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 type:
- laboratory study
- Substrate type:
- artificial soil
- Limit test:
- no
- Total exposure duration:
- 56 d
- Remarks:
- Removement of adults after 28 days
- Test temperature:
- 18 - 22.5 deg C
- pH:
- Application rate
[mg test item/kg soil DW] pH-value
day 0 day 56
Control 5.42 6.13
Solv. Control 5.40 6.40
62.5 5.39 6.18
125 5.39 6.30
250 5.40 6.33
500 5.38 6.07
1000 5.42 6.14 - Moisture:
- Application rate
[mg test item/kg soil DW] Moisture [%] of dry weight
day 0 day 56
Control 35.6 42.5*
Solv. Control 35.1 45.8*
62.5 34.8 46.7*
125 35.5 46.8*
250 34.9 44.6*
500 35.2 45.2*
1000 34.6 44.6*
*Variation of the water content > 10 %, this deviation is considered to have no impact on the study - 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: 4
- No. of replicates per control: 4
- No. of replicates per vehicle control: 4
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): 68
- CEC: not determined
- Pretreatment of soil: none
- Storage (condition, duration): 8 days after preparation at test conditions in the dark
OTHER TEST CONDITIONS
- Photoperiod: 16 h light - 8 h dark
- Light intensity: 524 +/- 70 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
TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2 - Nominal and measured concentrations:
- Nominal application rates: 62.5 - 125 - 250 - 500 - 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
- 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 mortality of adult earthworms was observed in the controls as well as at all application rates after 28 days of exposure to the test item
- 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 body weights of the earthworms increased in the control and at the test item concentrations of 62.5, 125, 250, 500 and 1000 mg test item / kg soil dry weight throughout the first 28 days. The biomass did not differ statistically significant compared to the pooled control at all test item concentrations
- No. of offspring produced: The mean number of juveniles in the pooled control was 359. The average number of juveniles in the treated groups ranged between 288 and 365. Compared to the pooled control the reproduction of the earthworms did not statistically significantly differ at all test item application rates
- Behavioural abnormalities: No symptoms or changes in the behaviour of the earthworms were observed in the controls as well as at all application rates after 28 days of exposure to the test item - 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:
- Differences between the control and the solvent control were statistically analysed using t-tests. No statistical differences occurred between the controls concerning biomass and reproduction. Thus, both controls were pooled for further statistical analyses.
One Way Analysis of Variance (ANOVA) was carried out for the determination of statistically significant differences compared to
the control. A Normality Test and an Equal Variance Test were conducted prior to running the ANOVA. P-values for both Normality and Equal Variance Test are 0.05. The a-value for ANOVA test (acceptable probability of incorrectly concluding that there is a difference) is a = 0.05. - Validity criteria fulfilled:
- yes
- Conclusions:
- In this earthworm reproduction test according to OECD 222 the submission substance did not induce mortality or pathological symptoms of adult earthworms after an exposure to the test item concentrations of 62.5, 125, 250, 500 and 1000 mg/kg soil dry weight for 28 days. Also, the biomass increase of the adult earthworms after 28 days did not statistically significantly differ compared to the pooled control at all test item concentrations. In addition, after eight weeks of exposure the earthworm reproduction was not statistically significantly reduced at all applied test item concentrations.
Thus the No Observed Effect Concentration (NOEC) for the submission substance with respect to mortality, body weight and reproduction of Eisenia fetida fetida after 8 weeks of exposure in artificial soil was determined to be 1000 mg test item/kg soil dry weight. - Executive summary:
Effects of the submission substance on mortality, biomass and the reproductive potential of the earthworm species Eisenia fetida (Savigny) were determined compliant to GLP according to the guidelines DIN ISO 11268-2 (1998), OECD 222 (2004) and BBA (1994) (reliability category 1). The study was conducted under static conditions over 8 weeks with five different application rates (62.5 - 125 - 250 - 500 - 1000 mg test item/kg soil dry weight), which were mixed into artificial soil containing 10 % peat. 40 test organisms, with an individual weight of 0.30 to 0.60 g, were divided into 4 replicate groups with 10 test organisms per replicate.
After 28 days no mortality as well as no pathological symptoms or changes in the behaviour of adult earthworms were seen in the control, in the solvent control and at all tested application rates. The earthworm biomass increased in the control and at all application rates over 28 days without any statistically significant differences compared to the control. After further four weeks the reproduction rate (average number of juveniles) was 359 in the control (control and solvent control) and ranged between 288 and 365 at the test item application rates. Compared to the control, the reproduction rate did not statistically significantly differ at all application rates.
Overall, the NOEC of the submission substance concerning mortality, biomass and reproduction of earthworms was determined to be >= 1000 mg test item/kg soil dry weight (Table 2).
All validity criteria recommended by the test guidelines were fulfilled.
Summary of All Observed Effects in the Tested Doses
Effects
Test item
mg test item/kg soil dry weight
62.5
125
250
500
1000
Mortality of adult earthworms
No
No
No
No
No
Reduction of body weight
No
No
No
No
No
Reduced reproduction rate
(Number of juveniles)
No
No
No
No
No
Summary of all effects
No
No
No
No
No
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
Referenceopen allclose all
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 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
Solvent Control |
0 |
0 |
0 |
10 |
0 |
0 |
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) |
10/10 |
10/10 |
10/10 |
10/10 |
10/10 |
10/10 |
10/10 |
10/10 |
Solvent Control |
A) |
10/10 |
10/10 |
10/10 |
9/10 |
10/10 |
10/10 |
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 kg soil dry weight] |
Rep. |
Mean body weights per replicate |
Mean body weight change of earthworms |
||||
Test start |
28 days |
Per replicate |
Mean ± SD [%] |
Significance compared to pooled control with [%] data |
|||
[g] |
[g] |
[g] |
[%] |
||||
Control |
1 |
0.573 |
0.827 |
0.25 |
44.3 |
54.3 |
– |
2 |
0.548 |
0.805 |
0.26 |
46.9 |
|||
3 |
0.498 |
0.800 |
0.30 |
60.6 |
|||
4 |
0.526 |
0.830 |
0.30 |
57.8 |
|||
5 |
0.535 |
0.799 |
0.26 |
49.3 |
|||
6 |
0.494 |
0.819 |
0.33 |
65.8 |
|||
7 |
0.516 |
0.781 |
0.27 |
51.4 |
|||
8 |
0.520 |
0.822 |
0.30 |
58.1 |
|||
Solvent Control |
1 |
0.524 |
0.817 |
0.29 |
55.9 |
53.4 |
– |
2 |
0.555 |
0.805 |
0.25 |
45.0 |
|||
3 |
0.553 |
0.794 |
0.24 |
43.6 |
|||
4 |
0.504 |
0.869 |
0.36 |
72.4 |
|||
5 |
0.529 |
0.811 |
0.28 |
53.3 |
|||
6 |
0.535 |
0.788 |
0.25 |
47.3 |
|||
7 |
0.486 |
0.773 |
0.29 |
59.1 |
|||
8 |
0.519 |
0.780 |
0.26 |
50.3 |
|||
1000 |
1 |
0.518 |
0.790 |
0.27 |
52.5 |
48.0 |
No |
2 |
0.519 |
0.763 |
0.24 |
47.0 |
|||
3 |
0.565 |
0.831 |
0.27 |
47.1 |
|||
4 |
0.545 |
0.792 |
0.25 |
45.3 |
|||
5 |
0.495 |
0.715 |
0.22 |
44.4 |
|||
6 |
0.525 |
0.779 |
0.25 |
48.4 |
|||
7 |
0.525 |
0.743 |
0.22 |
41.5 |
|||
8 |
0.490 |
0.774 |
0.28 |
58.0 |
|||
Reproduction Rate (Number of Juveniles after 8 Weeks)
Application rate [mg test item/kg soil dry weight] |
Rep. |
Number of juveniles |
Mean ± SD |
CV [%] |
[%] of control |
Significance compared to pooled control |
Control |
1 |
340 |
333 ± 35.7 |
10.7 |
– |
– |
2 |
299 |
|||||
3 |
356 |
|||||
4 |
288 |
|||||
5 |
378 |
|||||
6 |
290 |
|||||
7 |
365 |
|||||
8 |
350 |
|||||
Solvent Control |
1 |
328 |
332 ± 56.3 |
17.0 |
– |
– |
2 |
293 |
|||||
3 |
290 |
|||||
4 |
272 |
|||||
5 |
339 |
|||||
6 |
455 |
|||||
7 |
340 |
|||||
8 |
335 |
|||||
1000 |
1 |
337 |
375 ± 25.0 |
6.68 |
113 |
Yes* |
2 |
354 |
|||||
3 |
362 |
|||||
4 |
395 |
|||||
5 |
361 |
|||||
6 |
410 |
|||||
7 |
390 |
|||||
8 |
392 |
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 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
Solvent Control |
0 |
0 |
0 |
10 |
0 |
0 |
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) |
10/10 |
10/10 |
10/10 |
10/10 |
10/10 |
10/10 |
10/10 |
10/10 |
Solvent Control |
A) |
10/10 |
10/10 |
10/10 |
9/10 |
10/10 |
10/10 |
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 kg soil dry weight] |
Rep. |
Mean body weights per replicate |
Mean body weight change of earthworms |
||||
Test start |
28 days |
Per replicate |
Mean ± SD [%] |
Significance compared to pooled control with [%] data |
|||
[g] |
[g] |
[g] |
[%] |
||||
Control |
1 |
0.573 |
0.827 |
0.25 |
44.3 |
54.3 |
– |
2 |
0.548 |
0.805 |
0.26 |
46.9 |
|||
3 |
0.498 |
0.800 |
0.30 |
60.6 |
|||
4 |
0.526 |
0.830 |
0.30 |
57.8 |
|||
5 |
0.535 |
0.799 |
0.26 |
49.3 |
|||
6 |
0.494 |
0.819 |
0.33 |
65.8 |
|||
7 |
0.516 |
0.781 |
0.27 |
51.4 |
|||
8 |
0.520 |
0.822 |
0.30 |
58.1 |
|||
Solvent Control |
1 |
0.524 |
0.817 |
0.29 |
55.9 |
53.4 |
– |
2 |
0.555 |
0.805 |
0.25 |
45.0 |
|||
3 |
0.553 |
0.794 |
0.24 |
43.6 |
|||
4 |
0.504 |
0.869 |
0.36 |
72.4 |
|||
5 |
0.529 |
0.811 |
0.28 |
53.3 |
|||
6 |
0.535 |
0.788 |
0.25 |
47.3 |
|||
7 |
0.486 |
0.773 |
0.29 |
59.1 |
|||
8 |
0.519 |
0.780 |
0.26 |
50.3 |
|||
1000 |
1 |
0.518 |
0.790 |
0.27 |
52.5 |
48.0 |
No |
2 |
0.519 |
0.763 |
0.24 |
47.0 |
|||
3 |
0.565 |
0.831 |
0.27 |
47.1 |
|||
4 |
0.545 |
0.792 |
0.25 |
45.3 |
|||
5 |
0.495 |
0.715 |
0.22 |
44.4 |
|||
6 |
0.525 |
0.779 |
0.25 |
48.4 |
|||
7 |
0.525 |
0.743 |
0.22 |
41.5 |
|||
8 |
0.490 |
0.774 |
0.28 |
58.0 |
|||
Reproduction Rate (Number of Juveniles after 8 Weeks)
Application rate [mg test item/kg soil dry weight] |
Rep. |
Number of juveniles |
Mean ± SD |
CV [%] |
[%] of control |
Significance compared to pooled control |
Control |
1 |
340 |
333 ± 35.7 |
10.7 |
– |
– |
2 |
299 |
|||||
3 |
356 |
|||||
4 |
288 |
|||||
5 |
378 |
|||||
6 |
290 |
|||||
7 |
365 |
|||||
8 |
350 |
|||||
Solvent Control |
1 |
328 |
332 ± 56.3 |
17.0 |
– |
– |
2 |
293 |
|||||
3 |
290 |
|||||
4 |
272 |
|||||
5 |
339 |
|||||
6 |
455 |
|||||
7 |
340 |
|||||
8 |
335 |
|||||
1000 |
1 |
337 |
375 ± 25.0 |
6.68 |
113 |
Yes* |
2 |
354 |
|||||
3 |
362 |
|||||
4 |
395 |
|||||
5 |
361 |
|||||
6 |
410 |
|||||
7 |
390 |
|||||
8 |
392 |
Mortality of Adult Earthworms in [%] after 28 Days of Exposure
Application rate [mg test item / kg soil dry weight] |
Mortality [%] |
||||
Replicate 1 |
Replicate 2 |
Replicate 3 |
Replicate 4 |
||
Control |
0 |
0 |
0 |
0 |
|
Solvent Control |
0 |
0 |
0 |
0 |
|
62.5 |
0 |
0 |
0 |
0 |
|
125 |
0 |
0 |
0 |
0 |
|
250 |
0 |
0 |
0 |
0 |
|
500 |
0 |
0 |
0 |
0 |
|
1000 |
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 |
||
Control |
a) |
10/10 |
10/10 |
10/10 |
10/10 |
Solvent Control |
a) |
10/10 |
10/10 |
10/10 |
10/10 |
62.5 |
a) |
10/10 |
10/10 |
10/10 |
10/10 |
125 |
a) |
10/10 |
10/10 |
10/10 |
10/10 |
250 |
a) |
10/10 |
10/10 |
10/10 |
10/10 |
500 |
a) |
10/10 |
10/10 |
10/10 |
10/10 |
1000 |
a) |
10/10 |
10/10 |
10/10 |
10/10 |
Body Weight Changes of the Adult Earthworms
Application rate |
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 |
0.481 |
0.541 |
0.06 |
12.5 |
0.10 ± 0.03 |
22.5 ± 6.42 |
- |
2 |
0.448 |
0.541 |
0.09 |
20.8 |
||||
3 |
0.439 |
0.565 |
0.13 |
28.7 |
||||
4 |
0.453 |
0.552 |
0.10 |
21.9 |
||||
Solvent Control |
1 |
0.441 |
0.570 |
0.13 |
29.3 |
|||
2 |
0.479 |
0.563 |
0.08 |
17.5 |
||||
3 |
0.447 |
0.583 |
0.14 |
30.4 |
||||
4 |
0.449 |
0.534 |
0.09 |
18.9 |
||||
62.5 |
1 |
0.479 |
0.593 |
0.11 |
23.8 |
0.13 ± 0.03 |
28.2 ± 7.74 |
No |
2 |
0.444 |
0.600 |
0.16 |
35.1 |
||||
3 |
0.451 |
0.605 |
0.15 |
34.2 |
||||
4 |
0.452 |
0.540 |
0.09 |
19.5 |
||||
125 |
1 |
0.451 |
0.580 |
0.13 |
28.6 |
0.13 ± 0.01 |
28.2 ± 3.63 |
No |
2 |
0.451 |
0.565 |
0.11 |
25.3 |
||||
3 |
0.478 |
0.601 |
0.12 |
25.7 |
||||
4 |
0.437 |
0.582 |
0.15 |
33.2 |
||||
250 |
1 |
0.477 |
0.575 |
0.10 |
20.5 |
0.12 ± 0.03 |
27.1 ± 6.57 |
No |
2 |
0.448 |
0.594 |
0.15 |
32.6 |
||||
3 |
0.453 |
0.554 |
0.10 |
22.3 |
||||
4 |
0.444 |
0.590 |
0.15 |
32.9 |
||||
500 |
1 |
0.444 |
0.581 |
0.14 |
30.9 |
0.12 ± 0.03 |
27.3 ± 7.81 |
No |
2 |
0.442 |
0.581 |
0.14 |
31.4 |
||||
3 |
0.454 |
0.596 |
0.14 |
31.3 |
||||
4 |
0.475 |
0.549 |
0.07 |
15.6 |
||||
1000 |
1 |
0.454 |
0.599 |
0.15 |
31.9 |
0.14 ± 0.02 |
29.9 ± 5.07 |
No |
2 |
0.445 |
0.606 |
0.16 |
36.2 |
||||
3 |
0.480 |
0.606 |
0.13 |
26.3 |
||||
4 |
0.441 |
0.553 |
0.11 |
25.4 |
||||
Reproduction Rate (Number of Juveniles after 8 Weeks)
Application rate |
Rep. |
Number of juveniles |
Mean ± SD |
CV [%] |
[%] of pooled control |
Significance compared to pooled control |
Control |
1 |
359 |
359 ± 49.6 |
13.8 |
- |
- |
2 |
338 |
|||||
3 |
456 |
|||||
4 |
323 |
|||||
Solvent Control |
1 |
354 |
||||
2 |
388 |
|||||
3 |
287 |
|||||
4 |
365 |
|||||
62.5 |
1 |
338 |
365 ± 28.5 |
7.81 |
102 |
No |
2 |
356 |
|||||
3 |
360 |
|||||
4 |
405 |
|||||
125 |
1 |
284 |
355 ± 60.2 |
17.0 |
98.9 |
No |
2 |
327 |
|||||
3 |
393 |
|||||
4 |
415 |
|||||
250 |
1 |
378 |
347 ± 88.3 |
25.5 |
96.6 |
No |
2 |
442 |
|||||
3 |
334 |
|||||
4 |
232 |
|||||
500 |
1 |
303 |
288 ± 49.1 |
17.1 |
80.3 |
No |
2 |
250 |
|||||
3 |
351 |
|||||
4 |
248 |
|||||
1000 |
1 |
344 |
346 ± 83.8 |
24.2 |
96.4 |
No |
2 |
385 |
|||||
3 |
424 |
|||||
4 |
230 |
Mortality of Adult Earthworms in [%] after 28 Days of Exposure
Application rate [mg test item / kg soil dry weight] |
Mortality [%] |
||||
Replicate 1 |
Replicate 2 |
Replicate 3 |
Replicate 4 |
||
Control |
0 |
0 |
0 |
0 |
|
Solvent Control |
0 |
0 |
0 |
0 |
|
62.5 |
0 |
0 |
0 |
0 |
|
125 |
0 |
0 |
0 |
0 |
|
250 |
0 |
0 |
0 |
0 |
|
500 |
0 |
0 |
0 |
0 |
|
1000 |
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 |
||
Control |
a) |
10/10 |
10/10 |
10/10 |
10/10 |
Solvent Control |
a) |
10/10 |
10/10 |
10/10 |
10/10 |
62.5 |
a) |
10/10 |
10/10 |
10/10 |
10/10 |
125 |
a) |
10/10 |
10/10 |
10/10 |
10/10 |
250 |
a) |
10/10 |
10/10 |
10/10 |
10/10 |
500 |
a) |
10/10 |
10/10 |
10/10 |
10/10 |
1000 |
a) |
10/10 |
10/10 |
10/10 |
10/10 |
Body Weight Changes of the Adult Earthworms
Application rate |
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 |
0.481 |
0.541 |
0.06 |
12.5 |
0.10 ± 0.03 |
22.5 ± 6.42 |
- |
2 |
0.448 |
0.541 |
0.09 |
20.8 |
||||
3 |
0.439 |
0.565 |
0.13 |
28.7 |
||||
4 |
0.453 |
0.552 |
0.10 |
21.9 |
||||
Solvent Control |
1 |
0.441 |
0.570 |
0.13 |
29.3 |
|||
2 |
0.479 |
0.563 |
0.08 |
17.5 |
||||
3 |
0.447 |
0.583 |
0.14 |
30.4 |
||||
4 |
0.449 |
0.534 |
0.09 |
18.9 |
||||
62.5 |
1 |
0.479 |
0.593 |
0.11 |
23.8 |
0.13 ± 0.03 |
28.2 ± 7.74 |
No |
2 |
0.444 |
0.600 |
0.16 |
35.1 |
||||
3 |
0.451 |
0.605 |
0.15 |
34.2 |
||||
4 |
0.452 |
0.540 |
0.09 |
19.5 |
||||
125 |
1 |
0.451 |
0.580 |
0.13 |
28.6 |
0.13 ± 0.01 |
28.2 ± 3.63 |
No |
2 |
0.451 |
0.565 |
0.11 |
25.3 |
||||
3 |
0.478 |
0.601 |
0.12 |
25.7 |
||||
4 |
0.437 |
0.582 |
0.15 |
33.2 |
||||
250 |
1 |
0.477 |
0.575 |
0.10 |
20.5 |
0.12 ± 0.03 |
27.1 ± 6.57 |
No |
2 |
0.448 |
0.594 |
0.15 |
32.6 |
||||
3 |
0.453 |
0.554 |
0.10 |
22.3 |
||||
4 |
0.444 |
0.590 |
0.15 |
32.9 |
||||
500 |
1 |
0.444 |
0.581 |
0.14 |
30.9 |
0.12 ± 0.03 |
27.3 ± 7.81 |
No |
2 |
0.442 |
0.581 |
0.14 |
31.4 |
||||
3 |
0.454 |
0.596 |
0.14 |
31.3 |
||||
4 |
0.475 |
0.549 |
0.07 |
15.6 |
||||
1000 |
1 |
0.454 |
0.599 |
0.15 |
31.9 |
0.14 ± 0.02 |
29.9 ± 5.07 |
No |
2 |
0.445 |
0.606 |
0.16 |
36.2 |
||||
3 |
0.480 |
0.606 |
0.13 |
26.3 |
||||
4 |
0.441 |
0.553 |
0.11 |
25.4 |
||||
Reproduction Rate (Number of Juveniles after 8 Weeks)
Application rate |
Rep. |
Number of juveniles |
Mean ± SD |
CV [%] |
[%] of pooled control |
Significance compared to pooled control |
Control |
1 |
359 |
359 ± 49.6 |
13.8 |
- |
- |
2 |
338 |
|||||
3 |
456 |
|||||
4 |
323 |
|||||
Solvent Control |
1 |
354 |
||||
2 |
388 |
|||||
3 |
287 |
|||||
4 |
365 |
|||||
62.5 |
1 |
338 |
365 ± 28.5 |
7.81 |
102 |
No |
2 |
356 |
|||||
3 |
360 |
|||||
4 |
405 |
|||||
125 |
1 |
284 |
355 ± 60.2 |
17.0 |
98.9 |
No |
2 |
327 |
|||||
3 |
393 |
|||||
4 |
415 |
|||||
250 |
1 |
378 |
347 ± 88.3 |
25.5 |
96.6 |
No |
2 |
442 |
|||||
3 |
334 |
|||||
4 |
232 |
|||||
500 |
1 |
303 |
288 ± 49.1 |
17.1 |
80.3 |
No |
2 |
250 |
|||||
3 |
351 |
|||||
4 |
248 |
|||||
1000 |
1 |
344 |
346 ± 83.8 |
24.2 |
96.4 |
No |
2 |
385 |
|||||
3 |
424 |
|||||
4 |
230 |
Description of key information
PR112
Under the conditions of this study, Pigment Red 112 did not induce mortality or pathological symptoms of adult earthworms after an exposure to the test item concentration of 1000 mg test item/kg soil dry weight for 28 days. 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 was not statistically significantly reduced, but increased 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/ kg soil dry weight.
PO38
In this earthworm reproduction test according to OECD 222 the submission substance did not induce mortality or pathological symptoms of adult earthworms after an exposure to the test item concentrations of 62.5, 125, 250, 500 and 1000 mg/kg soil dry weight for 28 days. Also, the biomass increase of the adult earthworms after 28 days did not statistically significantly differ compared to the pooled control at all test item concentrations. In addition, after eight weeks of exposure the earthworm reproduction was not statistically significantly reduced at all applied test item concentrations.
Thus the No Observed Effect Concentration (NOEC) for the submission substance with respect to mortality, body weight and reproduction of Eisenia fetida fetida after 8 weeks of exposure in artificial soil was determined to be 1000 mg test item/kg soil dry weight.
Key value for chemical safety assessment
- Long-term EC10, LC10 or NOEC for soil macroorganisms:
- 1 000 mg/kg soil dw
Additional information
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.
