A mixture of: 2-ethylhexyl mono-D-glucopyranoside; 2-ethylhexyl di-D-glucopyranoside

Toxicity to soil macroorganisms except arthropods

Link to relevant study record(s)

Reference

Endpoint:

toxicity to soil macroorganisms except arthropods: long-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

December 17th 2014- February 12th 2015

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

OECD Guideline 222 (Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei))

Deviations:

yes

Remarks:

(The room temperature punctually reached 22.5°C(less than one hour at day 47)).

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
PHYSICO-CHEMICAL PROPERTIES
- Water solubility (under test conditions): soluble in cold water
-Water content : 1.5 %

Analytical monitoring:

not required

Vehicle:

no

Details on preparation and application of test substrate:

- Method of mixing into soil
The test item “Reaction mass of 2 ethylhexyl mono D-glucopyranoside, 2-ethylhexyl di-D-glucopyranoside” is soluble in water.
A stock solution of the test item was prepared in deionised water at 3 g/L.
The intended test item concentrations in soil were achieved by adding the corresponding volume of test item stock solution in the desired mass of artificial soil and mixing it for 5 minutes.
For the lower concentrations (29.8 to 312.5 mg/L), the corresponding volume of test item stock solution was diluted in deionised water in order to reach 250 ml and then added to the artificial soil and mixed for 5 minutes. This step allows homogeneous mixing of the test item in the artificial soil.
The final moisture content in the soils was then adjusted by adding deionised water. The soils have been mixed again for 5 minutes.

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

TEST ORGANISM
- Common name: Eisenia fetida
- Source: breeding in test facility
- Age at test initiation: Between 5 and 7 months
- Weight at test initiation (mean and range, SD):
At the initiation of the test, 30 worms were randomly sampled from the test population and weighted :
Mean value : 404.2 mg
Minimal value : 324.1 mg
Maximal value : 580.9 mg
Standard deviation : 60.4

ACCLIMATION
- Acclimation period: 24 hours in artificial soil
- Acclimation conditions (same as test or not): same as the test
- Health during acclimation (any mortality observed): no mortality

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

no

Total exposure duration:

56 d

Post exposure observation period:

None

Test temperature:

Stabilized in the range of 18 to 22°C during the exposure period, the room temperature reached 22.5°C at day 47, during less than one hour.

pH:

See Table 2 for pH measurments at day 0, day 28 and day 56 for control and test concentrations.

Moisture:

See Table 3 for moisture measurments at day 0, day 28 and day 56 for control and test concentrations.

Details on test conditions:

TEST SYSTEM
- Test container (material, size): Glass containers (177 cm2 X 7 cm)
- Amount of soil or substrate: around 500g dry mass corresponding to a layer of 5-6 cm
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 4
- No. of replicates per control: 8

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
Artificial soil prepared according to the guideline recommandations
- Composition :
10% blond sphagnum peat air dried and finely ground
20% kaolin (acid washed powder)
70% air dried quartz sand (HN38) containing 94.9% of particles between 50 and 200µm
0.3% Calcium carbonate
- Maximum water holding capacity (in % dry weigth): 78.8
- pH measured with KCl : 5.9

OTHER TEST CONDITIONS
- Photoperiod: 16 hours light - 8 hours darkness
- Light intensity: range from 415 to 782 Lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Mortality and growth of adults after 4 weeks exposure
- Reproductive output (number of juveniles per test condition) after 8 weeks exposure

VEHICLE CONTROL PERFORMED: no

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.6
- Range finding study : Mortality of adults after 14 days exposure, 2 replicates per concentration
- Test concentrations( in mg/kg dry mass): 0.1 - 1.0 - 10 - 100 - 1 000
- Results used to determine the conditions for the definitive study: 15 % mortality after 14 days at 1 000 mg/kg (dry mass)

Nominal and measured concentrations:

Nominal test concentrations (in mg/kg dry mass): 29.8 - 47.7 - 76.3 - 122.1 - 195.3 - 312.5 - 500.0 - 800.0
No test concentration measurments were performed

Reference substance (positive control):

yes

Remarks:

Benomyl at 1 and 3 mg/kg (dry mass)

Key result

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

500 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks:

adults

Duration:

28 d

Dose descriptor:

LOEC

Effect conc.:

800 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks:

adults

Duration:

56 d

Dose descriptor:

NOEC

Effect conc.:

800 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Remarks on result:

other: Highest tested concentration

Details on results:

- Mortality of adults at end of 28 days exposure : 2.5 % at 29.8 mg/kg dry masse, 12.5 % at 800 mg/kg dry mass (see table 4 for details)
- Morphological abnormalities: None
- Behavioural abnormalities: None

- Total mass of adults at beginning of test: See table 4
- Changes in body weigth of live adults (% of initial weight) at end of exposure period: See table 4
- No. of offspring produced: See table 6

Results with reference substance (positive control):

- Reference item :
Two concentrations of Benomyl were prepared (1 and 3 mg/kg dry mass) and four replicates per concentration were performed.
Considering the use of solvent (acetone) to prepare refernce substance concentrations, foru solvent controls were prepared

- Relevant effect levels: at 3 mg/kg dry mass
*Mean change of the body fresh weight = 19.7% versus 88.4% for solvent control , corresponding to 77.8% inhibition compared to solvent control
*Reproductive output = 0 juveniles per replicate, corresponding to 100% reproduction inhibition compared to solvent control

Reported statistics and error estimates:

For the test performed with the test item, significant differences in changes in mean body weight and reproductive output were calculated using one way ANOV (p-value <5) followed by the Bonferroni post-hoc test (p-value <5), after verification of the normality and the homogeneity of data ( Toxstat 3.5, Western Inc and D. Gulley, University of Wyoming).
For the test performed with thereference item, significant differences in changes in mean body weight and reproductive output were calculated using one way ANOVA (p-value <5) followed by theDunett's test (p-value <5), after verification of the normality and the homogeneity of data ( Toxstat 3.5, Western Inc and D. Gulley, University of Wyoming).

RESULTS

1 - REFERENCE ITEM
After exposure to the artificial soil containing 1.0 mg/kg (dry mass) of reference item, the fresh body weight increase (7.4% relative to the control) and reproduction output (-22.7% relative to the control) were not statistically different by comparison with the data recorded for the worms exposed to the solvent control soil.
Significant differences were observed for the organisms exposed to the reference item at 3.0 mg/kg of soil (dry mass). The following observations were made:
- A statistically significant reduction of the body fresh weight increase (-77.8%
relative to the solvent control)
- A total inhibition of reproduction.

These results, summarised in Table 4, confirmed the appropriate sensitivity of the test organisms.

Table 4: Results of the reproduction test performed with the reference item

	Solvent control	Reference item concentration (mg/kg dry mass)
		1.0	3.0
Mortality after 4 weeks exposure	0.0%	0.0%	0.0%
Mean change in body fresh weight after 4 weeks exposure	88.4%	94.9%	19.7%*
Variation of body weight (compared to control)	/	7.4%	-77.8*
Reproductive output (juveniles per replicate)	90.5	70.0	0.0*
Variation of reproductive output (compared to the control)	/	-22.7%	-100.0%*

*Results statistically significant (one-way ANOVA (p-value < 0.05) followed by the Dunnett’s post-hoc test (p-value < 0.05)), Toxstat 3.5, Western Inc. and D. Gulley, University of Wyoming.

The experiment on the reference item was designed to encompass the EC50 value and to check that this value was included in the appropriate range of concentration. Twoconcentrations were tested: 1 and 3 mg benomyl /kg of soil (dry mass) and compared to a solvent control.
Each solvent control replicate produced more than 30 juveniles by the end of the test and no mortality was recorded in such control. The coefficient of variation of the reproduction endpoint (e.g. 30.9%) exceeded slightly the 30% threshold require by the OECD Guideline for testing of chemicals n°222 (see paragraph V. Validity Criteria). However in such case (i.e. use of four replicate of solvent control), the suitability of this validity criterion needs to be discussed and can’t be directly transposed as the CV of reproduction for control is generally calculated on six to eight control replicates (depending on the test design for NOEC, ECx determination or combined approach). Given that (i) that the CV of reproduction calculated for the test item assay performed during the same period didn’t exceed 27%, and (ii) that the results obtained for reproduction in both reference item (mean = 90.5; SD = 27.9) and test item assay (mean = 106.3; SD = 28.6) are similar, it can therefore be assumed that this slightly higher CV for the solvent control didn’t invalid the results of the test with reference item.

 

2 - TEST ITEM CONCENTRATIONS PH AND WATER CONTENTS MEASUREMENTS
The pH values and the water content in control soil and test item concentrations are summarised in the Table 5.
Table 5: pH and water content measured in test concentrations and in controls during the study

Nominal test item concentration (mg/kg dry mass) pH Water content (%) Day 0 Day 28 Day 56 Day 0 Day 28 Day 56 Variation between Day 0 and Day 56  Control 5.6 6.8  7.1  42.8  43.3  45.4  6%  29.8 5.5  6.6  7.1   43.2  42.6  45.6 6%  47.7 5.5  6.7  7.0   42.9  43.2  46.5 8%  76.3 5.6  6.6  7.0   43.1  43.7  43.0 0%  122.1 5.6  6.8  6.9   42.9  41.7  44.5 4%  195.3 5.5 6.9  6.9   42.6  45.4  45.7 7%  312.5 5.5  6.9  7.0   43.0  43.0  47.1 9%  500.0 5.5  6.8  6.9   42.8  42.5  46.8 9%  800.0 5.5  6.8  6.9   42.7  42.9  46.8 9%

 

The water content in soil didn’t vary more than 9 % between the beginning and the end of the test.

 

3 - MORTALITY AND GROWTH OF ADULTS EARTHWORMS
Results obtained for fresh body weight variations and mortality are presented in Tables 6.
After four weeks of exposure, no significant increase of the mortality was observed up to 500.0 mg/kg of soil (dry mass) and no mortality was observed in the control replicates. At 800 mg/kg of soil (dry mass), a slight but statistically significant increase of the earthworms mortality was recorded (12.5% relative to the control). It should be noted that mortality (including missing worms) was observed only in two of the four replicates and consequently, these results should be considered cautiously. No other adverse effects were observed.
The mean fresh body weight of the adult worms increased in the control (101.8%) and in all test concentrations (between 101.6 and 112.2%, depending on the test item concentration). These increases correspond to variations of -0.2% to 10.2% relative to the control condition (depending on the test item concentration). No statistically significant effect of the test item was observed on the growth of the adult earthworms whatever the test item concentration.

 

Table 6 (4): Mass of adults and changes in the body weight between the beginning and the end of the 28 days exposure period.

		Day 0	Day 28			Changes in body weight (% of initial weight)
Nominal test item concentrations (mg/kg dry mass)		Earthworms total body weight (mg)	Number of surviving earthworms	Earthworms total body weight (mg)	Mean body weight / earthworm (mg)
Control	1	4 046	10	7 701	770.1	90.3
	2	3 880	10	7 492	749.2	93.1
	3	3 601	10	7 050	705.0	95.8
	4	3 455	10	7 746	774.6	124.2
	5	3 874	10	7 578	757.8	95.6
	6	3 649	10	7 445	744.5	104.0
	7	3 720	10	7 178	717.8	93.0
	8	3 415	10	7 467	746.7	118.7
Mean SD (s)		3 705.0 205.0	10 /	7 457.1 224.3	745.7 22.4	101.8 12.0
29.8	1	4 029	10	8 316	831.6	106.4
	2	3 717	9	6 619	735.4	97.9
	3	3 582	10	8 022	802.2	124.0
	4	3 759	10	7 537	753.7	100.5
Mean SD (s)		3 771.8 162.3	9.75 /	7 623.5 643.2	780.7 38.2	107.2 10.2
47.7	1	3 600	10	7 517	751.7	108.8
	2	3 609	10	7 685	768.5	112.9
	3	3 866	10	8 099	809.9	109.5
	4	3 552	10	7 733	773.3	117.7
Mean SD (s)		3 656.8 122.7	10 /	7 758.5 212.3	775.9 21.2	112.2 3.5
76.3	1	3 838	10	7 839	783.9	104.2
	2	3 777	10	7 560	756.0	100.2
	3	3 733	10	7 169	716.9	92.0
	4	3 723	10	7 861	786.1	111.1
Mean SD (s)		3 767.8 45.4	10 /	7 607.3 279.5	760.7 27.9	101.9 6.9
122.1	1	3 635	10	7 743	774.3	113.0
	2	3 841	10	7 834	783.4	104.0
	3	3 842	10	7 981	798.1	107.7
	4	3 633	10	7 374	737.4	103.0
Mean SD (s)		3 737.8 103.8	10 /	7 733.0 224.0	773.3 22.4	106.9 3.9
195.3	1	3 793	10	7 459	745.9	96.7
	2	3 894	10	7 351	735.1	88.8
	3	3 691	10	8 001	800.1	116.8
	4	3 421	10	7 384	738.4	115.8
Mean SD (s)		3 699.8 176.2	10 /	7 548.8 264.0	754.9 26.4	104.5 12.1
312.5	1	3 856	10	7 332	733.2	90.1
	2	3 747	10	7 868	786.8	110.0
	3	3 872	10	7 856	785.6	102.9
	4	3 603	10	7 548	754.8	109.5
Mean SD (s)		3 769.5 107.5	10 /	7 651.0 224.4	765.1 22.4	103.1 8.0
500.0	1	3 611	10	7 334	733.4	103.1
	2	3 907	10	7 843	784.3	100.7
	3	3 611	10	7 650	765.0	111.9
	4	3 486	10	7 688	768.8	120.5
Mean SD (s)		3 653.8 154.9	10 /	7 628.8 184.9	762.9 18.5	109.1 7.8
800.0	1	3 987	10	7 459	745.9	87.1
	2	3 692	7	5 098	728.3	97.3
	3	3 441	8	5 773	721.6	109.7
	4	3 506	10	7 443	744.3	112.3
Mean SD (s)		3 656.5 211.9	8.8* /	6 443.3 1035.6	735.0 10.4	101.6 10.1

*Results statistically significant (one-way ANOVA (p-value < 0.05) followed by the Bonferroni post-hoc t-test (p-value < 0.05)), Toxstat 3.5, Western Inc. and D. Gulley, University of Wyoming.

 

4 - REPRODUCTIVE OUTPUT
The total number of living offspring produced by adults is shown in Table 7. In the control, an average of 106.3 juveniles (SD = 28.6) per test vessels was recorded. Results obtained after exposure to the test item didn’t show any concentration-effect relationship. Except for the treatment at 312 mg/kg (dry soil), the reproduction was always lower by comparison with the control (ranged from -15.1% to -38.4% relative to the control).
However, according to the statistical analyses (one-way ANOVA followed by the Bonferroni post-hoc t-test (p value < 0.05)), these differences are not significant.

Table 7: Number of juveniles per test vessel and variation of reproductive output in test item concentrations

Nominal test item concentrations (mg/kg dry mass)		Number of juveniles	CV (%)	Variation of reproductive output (%)
Control	1	162	26.9	/
	2	116
	3	121
	4	70
	5	88
	6	105
	7	118
	8	70
Mean SD (s)		106.3 28.6
29.8	1	71	18.8	-38.4
	2	82
	3	49
	4	60
Mean SD (s)		65.5 12.3
47.7	1	93	18.6	-30.1
	2	54
	3	74
	4	76
Mean SD (s)		74.3 13.8
76.3	1	90	15.9	-15.1
	2	100
	3	104
	4	67
Mean SD (s)		90.3 14.4
122.1	1	51	35.2	-23.1
	2	110
	3	111
	4	55
Mean SD (s)		81.8 28.8
195.3	1	94	11.1	-16
	2	103
	3	78
	4	82
Mean SD (s)		89.3 9.9
312.5	1	134	32.5	9.4
	2	52
	3	149
	4	130
Mean SD (s)		116.3 37.8
500.0	1	90	25.7	-36.2
	2	76
	3	62
	4	43
Mean SD (s)		67.8 17.4
800.0	1	113	39.4	-36.5
	2	59
	3	49
	4	49
Mean SD (s)		67.5 26.6

 

 

 

Validity criteria fulfilled:

yes

Remarks:

In control conditions: Each replicate produced more than 30 juveniles by the end of the test, the coefficient of variation was lower than 30% (26.9%) and the adult mortality over the initial 4 weeks didn't exceed 10 % ( 0%)

Conclusions:

Effect of the test item “Reaction mass of 2 ethylhexyl mono D-glucopyranoside, 2-ethylhexyl di-D-glucopyranoside” on earthworm reproduction was assessed by testing eight concentrations: 29.8 – 47.7 – 76.3 – 122.1 – 195.3 – 312.5 – 500.0 – 800.0 mg/kg.
The results obtained (summarised in Table 8) have shown a decrease of the reproduction when the worms were exposed to the test item, but no concentration-effect relationship was determined. Moreover, according to the statistical analyses (one-way ANOVA followed by the Bonferroni post-hoc t-test (p value < 0.05)), these differences in worms reproduction cannot be considered as significant. Taking altogether the mortality, the growth and the reproduction of the exposed worms, the highest concentration of the test item “Reaction mass of 2 ethylhexyl mono D-glucopyranoside, 2-ethylhexyl di-D-glucopyranoside” without statistically significant inhibitory effect on Eisenia fetida was determined to be, at least, 500 mg/kg of soil (dry mass). At 800 mg/kg of soil (dry mass), a statistically slight increase of the worms mortality was observed (12.5% relative to the control), while no effect on worms growth and reproduction was reported. However, this increase in worms mortality should be considered cautiously as missing worms were observed only for tow of the four test vessel replicates.
According to these results the NOEC value for worms mortality is therefore 500 mg/kg (dry mass) and the NOEC value for worms reproduction is 800 mg/kg (dry mass), the highest tested concentration.

Executive summary:

TEST PURPOSE

Determination of the effect of the test item "Reaction mass of 2 ethylhexyl mono D-glucopyranoside, 2-ethylhexyl di-D-glucopyranoside" on the reproductive output of the earthworm Eisenia fetida. The test was carried out according to the procedure described in the OECD Guideline for testing of chemicals n°222 "Earthworm Reproduction test", in conformity with the clauses recommended in the study plan of October 28th 2014, signed by the sponsor on November 6th 2014 and the study plan amendment of December 08th 2014, signed by the sponsor on December 11th 2014, signed by the sponsor on December 11th 2014.

 

GUIDELINE

OECD Guideline for testing of chemicals n°222 "Earthworm Reproduction test" (Adopted April 13th 2014).

 

TEST SYSTEM

Eisenia fetida

 

TEST ITEM

"Reaction mass of 2 ethylhexyl mono D-glucopyranoside, 2-ethylhexyl di-D-glucopyranoside"

 

TEST CONCENTRATION

Nominal concentrations: 29.8 - 47.7 - 76.3 - 122.1 - 195.3 - 312.5 - 500.0 - 800.0 mg/kg of soil (dry mass)

 

DURATION

56 days

 

RESULTS

Effects on earthworms exposed to the test item “Reaction mass of 2 ethylhexyl mono D-glucopyranoside, 2-ethylhexyl di-D-glucopyranoside”are summarised in Table 1 and Table 2.

After 4 weeks of exposure, no statistically significant increase of the worm mortality was recorded up to 500 mg/kg of soil (dry mass).

At 800 mg/kg of soil (dry mass) a mortality of 12.5% relative to the control was observed. According to the statistical analyses (one-way ANOVA followed by the Bonferroni post-hoc t-test (p value < 0.05)), this slight increase of the mortality was found to be significant. However, this result should be considered cautiously as missing worms were observed only for two of the four test vessel replicates.

Whatever the concentration tested, the increase of the fresh body weight was similar to the control group and no statistically significant inhibition was observed. The variation of the body fresh weight compared to the control ranged from -0.2 to 10.2%.

Concerning the reproduction, results obtained after exposure to the test item didn’t show any concentration-effect relationship. Except for the treatment at 312.5 mg/kg (dry soil), the reproduction was always lower by comparison with the control (ranged from -15.1% to -38.4% relative to the control). However, according to the statistical analyses (one-way ANOVA followed by the Bonferroni post-hoc t-test (p value < 0.05)), these differences are not significant.

Taking altogether the mortality, the growth and the reproduction of the exposed worms, the highest concentration of the test item“Reaction mass of 2 ethylhexyl mono D-glucopyranoside, 2-ethylhexyl di-D-glucopyranoside”without statistically significant inhibitory effect onEisenia fetida was determined to be, at least, 500 mg/kg of soil (dry mass). According to these results, the NOEC value for mortality is therefore 500 mg/kg (dry mass) and the NOEC value for reproduction is 800 mg/kg (dry mass), highest tested concentration.

Concerning the test with reference item (Table 2), statistically significant inhibition of the growth and the reproduction were observed for the organisms exposed to the reference item at 3.0 mg.kg of soil (dry mass).

 

Table 1: Effects on earthworms exposed to the test item “Reaction mass of 2 ethylhexyl mono D-glucopyranoside, 2-ethylhexyl di-D-glucopyranoside”

Parameters	Control	Test item (mg/kg dry mass)
		29.8	47.7	76.3	122.1	195.3	312.5	500	800
Mortality after 4 weeks exposure	0.0%	2.5%	0.0%	0.0%	0.0%	0.0%	0.0%	0.0%	12.5%*
Mean increase of body fresh weight after 4 weeks exposure	101.8%	107.2%	112.2%	101.9%	106.9%	104.5%	103.1%	109.1%	101.6%
Variation of body weight (compared to control)	/	5.3%	10.2%	0.1%	5.0%	2.6%	1.3%	7.1%	-0.2%
Reproductive output (juveniles per replicate)	106.3	65.5	74.3	90.3	81.8	89.3	116.3	67.8	67.5
Variation of reproductive output (compared to control)	/	-38.4%	-30.1%	-15.1%	-23.1%	-16%	9.4%	-36.2%	-36.5%
*Results statistically significant (one-way ANOVA (p-value < 0.05) followed by the Bonferroni post-hoc t-test (p-value < 0.05)), Toxstat 3.5, Western Inc. and D. Gulley, University of Wyoming.

Table 2: Effects on earthworms exposed to the reference substance (Benomyl)

Parameters	Solvent control	Reference Item
		1	3
Mortality after 4 weeks exposure	0.0%	0.0%	0.0%
Mean increase of body fresh weight after 4 weeks exposure	88.4%	94.0%	19.7%*
Variation of body weight (compared to control)	/	7.4%	-77.8%
Reproductive output (juveniles per replicate)	90.5	70.0	0.0*
Variation of reproductive output (compared to control)	/	-22.7%	-100.0%*

 

Description of key information

Effect of the test item “Reaction mass of 2 ethylhexyl mono D-glucopyranoside, 2-ethylhexyl di-D-glucopyranoside” on earthworm reproduction was assessed by testing eight concentrations: 29.8 – 47.7 – 76.3 – 122.1 – 195.3 – 312.5 – 500.0 – 800.0 mg/kg.
The results obtained (summarised in Table 8) have shown a decrease of the reproduction when the worms were exposed to the test item, but no concentration-effect relationship was determined. Moreover, according to the statistical analyses (one-way ANOVA followed by the Bonferroni post-hoc t-test (p value < 0.05)), these differences in worms reproduction cannot be considered as significant. Taking altogether the mortality, the growth and the reproduction of the exposed worms, the highest concentration of the test item “Reaction mass of 2 ethylhexyl mono D-glucopyranoside, 2-ethylhexyl di-D-glucopyranoside” without statistically significant inhibitory effect on Eisenia fetida was determined to be, at least, 500 mg/kg of soil (dry mass). At 800 mg/kg of soil (dry mass), a statistically slight increase of the worms mortality was observed (12.5% relative to the control), while no effect on worms growth and reproduction was reported. However, this increase in worms mortality should be considered cautiously as missing worms were observed only for tow of the four test vessel replicates.
According to these results the NOEC value for worms mortality is therefore 500 mg/kg (dry mass) and the NOEC value for worms reproduction is 800 mg/kg (dry mass), the highest tested concentration.

Key value for chemical safety assessment

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

500 mg/kg soil dw