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Toxicity to soil microorganisms

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Endpoint:
toxicity to soil microorganisms
Type of information:
experimental study
Adequacy of study:
key study
Study period:
23 Mar - 20 Apr 2004
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 216 (Soil Microorganisms: Nitrogen Transformation Test)
Version / remarks:
adopted 21 January 2000
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
Ministerium fur Umwelt, Raumordnung und Landwirtschaft des Landes Nordrhein-Westfalen, Düsseldorf, Germany
Analytical monitoring:
no
Vehicle:
no
Details on preparation and application of test substrate:
AMENDMENT OF SOIL
- Type of organic substrate: Lucerne-grass-green meal; source: Hoeveler Kraftfutter, Langenfeld, Germany

APPLICATION OF TEST SUBSTANCE TO SOIL
- Method: Sieved soil (2 mm) was treated with either 10 g ground quartz sand/kg dry weight soil (control) or a mixture of quartz sand and the test item (0.07 mg or 0.33 mg/kg dry weight soil). The samples were mixed with pulverized lucerne-grass-green meal (5 g/kg dry weight soil) and quartz sand in 3 liter polyethylene containers by rolling on a gyrowheel mixer (Elte 650, J. Engelsmann AG, Ludwigshafen, Germany) for 15 min at 50 rpm. In separate studies it was determined that mixing substances into soils as described gives a homogeneous and reproducible distribution (Blumenstock, 1989). Since the soil was not extracted and analyzed, values given for the active ingredients are nominal.

Test organisms (inoculum):
soil
Total exposure duration:
28 d
Test temperature:
20 ± 2 °C
Moisture:
40% of the maximum water holding capacity
Details on test conditions:
TEST SYSTEM
- Testing facility: Bayer CropScience AG
- Test container (type, material, size): 500 mL brown glass bottles and closed with parafilm.
- Amount of soil: 300 g dry weight
- No. of replicates per concentration: 3
- No. of replicates per control: 3

SOIL INCUBATION
- Method: bulk / series of individual subsamples
The soil was mixed and kept in a brown glass bottles closed with parafilm. It was held in the dark at 20 ± 2 °C and about 40% of the maximum water holding capacity (WHCmax).

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Geographical reference of sampling site (latitude, longitude): at a latitude of 51°4' north and a longitude of 6°55' east.
- History of site: The soil used is from Field plot F on the Bayer CropScience AG's experimental farm Laacherhof, Germany. The plot has been under grass and has not been treated with fertilizers since 1996.
- Vegetation cover: On 07 Mar 2000 the plot was plowed and then freshly planted with grass.
- Treatments with pesticides or fertilizers: Plant protection chemicals have not been used on this field since 1981.
- Accidental contamination: no
- Depth of sampling: 0-20 cm
-Soil texture "lehmiger Sand" (loamy sand)
- 2000 - 630 (µm): 4.0%
- 630 - 200 (µm): 52.5%
- 200 - 63 (µm): 21.6%
- 63 - 20 (µm): 6.9%
- 20 - 6.3 (µm): 4.3%
- 6.3 - 2.0 (µm): 4.4%
- 2.0 - 0 (µm): 6.3%
- Soil taxonomic classification: "lehmiger Sand" (loamy sand)
- pH (in water): 6.7
- Initial nitrate concentration for nitrogen transformation test (mg nitrate/kg dry weight): The loamy sand soil held 12.58 mg nitrate-N/kg dry weight soil
- Cation exchange capacity: 6.4 meq/100 g dry weight soil
- Pretreatment of soil: Sieved soil (2 mm) was treated with either 10 g ground quartz sand/kg dry weight soil (control) or a mixture of quartz sand and test item (0.07 mg or 0.33 mg/kg dry weight soil). The samples were mixed with pulverized lucerne-grass-green meal (5 g/kg dry weight soil) and quartz sand in 3 liter polyethylene containers by rolling on a gyrowheel mixer for 15 min at 50 rpm. Since the soil was not extracted and analyzed, values given for the active ingredients are nominal.
- Storage (condition, duration): stored at 4 ± 2 °C until used
- Initial microbial biomass as % of total organic C: 4.8%

DETAILS OF PREINCUBATION OF SOIL (if any): no preincubation

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Effects on non-target soil microorganisms are measured after 0, 7, 14, 28 days. Therefore the amount of Ammonium-N and Nitrate-N were determined as mg Nitrogen (N)/kg dry weight soil

VEHICLE CONTROL PERFORMED: no

RANGE-FINDING STUDY: no range-finding test was performed
Nominal and measured concentrations:
concentrations of 0.07 mg and 0.33 mg test item/kg dry weight soil
Reference substance (positive control):
yes
Duration:
28 d
Dose descriptor:
other: trigger value of 25% inhibition compared with the control
Effect conc.:
> 0.25 kg/ha
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
nitrate formation rate
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
>= 0.316 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
nitrate formation rate
Remarks on result:
other: Endpoint was not part of the report but derivable from the reported results
Duration:
28 d
Dose descriptor:
EC10
Effect conc.:
> 0.316 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
nitrate formation rate
Remarks on result:
other: Endpoint was not part of the report but derivable from the reported results
Details on results:
The substance caused no adverse effects (difference to control <25%, OECD 216) on the soil nitrogen transformation (expressed as NO3-N-production) at the end of the 28 d incubation period. The study was performed in a field soil at concentrations up to 0.316 mg a.s./kg soil dry weight, which are equivalent to application rates up to 0.25 test item/ha.
Results with reference substance (positive control):
- Results with reference substance valid? yes
- Relevant effect levels: 16 g NaCI/kg dry weight soil had a distinct and long-term (> 28 days) influence on microbial mineralization of nitrogen.
Reported statistics and error estimates:
All calculations were performed using Microsoft Excel 97. The % differences in the quantities of nitrate-N formed between control soils and treated soils were expressed as absolute values and determined as follows: ((treatment rates - control rates)/control rates) x 100 % = % difference. Rates were expressed in "mg nitrate-N/kg dry weight soil/day". Homogeneity of variances was determined by F-test (significance level 5%). Depending on the results of the F-Tests, the appropriate T-Tests were performed. In the T-Tests the mean values of nitrate-N/kg dry weight soil/time interval/day from control soils and treated soils were compared. The statistical calculations were carried out using Microsoft Excel 97 software.

During the 28-day test, in a soil amended with lucerne-grass-green-meal (5 g/kg), it was found that 0.07 mg test item/kg dry weight soil (equivalent to 0.05 kg product/ha) and the 5-fold overdose of the compound had no influence on nitrogen transformation in a loamy sand. In none of the time intervals analysed during the 28 day exposure the difference in the daily nitrate-N rates exceeds the trigger value of 25%.

The pH values show that the product did not cause a change in soil pH. Nitrite formation was not detected in any of the soil samples. The highest difference between nitrate-N rates per day of control and the one-foldtreated soil samples was 7% (14-28 days after soil treatment). The highest difference between nitrate-N rates per day of control and of the 5-foldoverdose-treated soil samples was 11% (0 -7 days after soil treatment). In this study, the highest coefficient of variation (CV) between nitrate-N concentration in replicate control samples was 6% (7 and 14 days after treatment) and thus did not exceed the recommended limit <15%.

Table 1: Nitrogen transformation: Indicidual values and mean values

Days after Treatment

Test item/ kg dry weight Soil

 

0 mg

0.07 mg

0.33 mg

 

mg Nitrogen (N) / kg dry weight Soil

0

 

Ammonium-N

Nitrate -N

 

Ammonium-N

Nitrate -N

 

Ammonium-N

Nitrate -N

0

R1

3.15

21.52

R4

3.27

21.33

R7

4.12

20.35

R2

3.38

21.32

R5

2.96

20.71

R8

2.80

20.27

R3

3.23

20.97

R6

2.96

20.36

R9

3.03

19.93

MV

3.25

21.27

MV

3.06

20.80

MV

3.32

20.19

SD

±0.12

± 0.28

SD

±0.18

±0.49

SD

±0.71

±0.22

CV

4%

1%

CV

 

 

CV

 

 

7

R1

1.63

6.11

R4

1.95

5.48

R7

2.33

6.73

R2

1.71

5.42

R5

2.49

6.35

R8

2.02

7.30

R3

2.06

5.93

R6

2.68

4.76

R9

1.95

5.37

MV

1.80

5.82

MV

2.37

5.53

MV

2.10

6.47

SD

± 0.23

±0.36

SD

±0.38

±0.80

SD

±0.21

±0.99

CV

13 %

6%

CV

 

 

CV

 

 

14

R1

1.48

11.28

R4

1.32

11.11

R7

1.75

12.46

R2

1.24

12.34

R5

1.32

11.50

R8

1.36

12.63

R3

1.36

10.99

R6

1.32

11.09

R9

1.36

12.17

MV

1.36

11.54

MV

1.32

11.23

MV

1.49

12.42

SD

±0.12

±0.71

SD

±0.00

±0.24

SD

±0.22

±0.23

CV

9%

6%

CV

 

 

CV

 

 

28

R1

1.95

25.63

R4

1.44

26.98

R7

1.40

26.08

R2

1.52

25.12

R5

1.32

25.47

R8

1.36

26.23

R3

1.36

25.57

R6

1.32

25.74

R9

1.36

26.56

MV

1.61

25.44

MV

1.36

26.07

MV

1.37

26.29

SD

±0.30

± 0.28

SD

±0.07

±0.81

SD

±0.02

±0.24

CV

19%

1 %

CV

 

 

CV

 

 

Samples of a loamy sand soil (0.7% org. C, pH (KCI) 5.6) were treated with the test item, amended with lucerne-grass-green meal (5 g/kg dry weight soil) and incubated in the dark at 20 ± 2 °C and 40% of the maximum water holding capacity.

Mean values (MV) and standard deviations (SO) from 3 soil samples (R). CV = Coefficient of Variation ((SO/MV)*100)

Table 2: Rates of nitrogen transformation / time interval: individual values and mean values

Time lnterval (Days)

Test item / kg dry weight Soil

 

0 mg

0.07 mg

0.33 mg

 

mg Nitrogen (N) / kg dry weight Soil / Time Interval

 

 

Nitrate-N

 

Nitrate-N

 

Nitrate-N

0-7

R1

-15.40

R4

-15.85

R7

-13.62

 

R2

-15.90

R5

-14.36

RB

-12.97

 

R3

-15.04

R6

-15.61

R9

-14.57

 

MV

-15.45

MV

-15.27

MV

-13.72

 

SD

±0.43

SD

±0.80

SD

±0.80

7-14

R1

5.16

R4

5.63

R7

5.73

 

R2

6.92

R5

5.15

RB

5.33

 

R3

5.06

R6

6.33

R9

6.80

 

MV

5.71

MV

5.70

MV

5.96

 

SD

±

SD

±0.59

SD

±0.76

14-28

R1

14.35

R4

15.88

R7

13.62

 

R2

12.78

R5

13.97

RB

13.59

 

R3

14.58

R6

14.66

R9

14.38

 

MV

13.90

MV

14.83

MV

13.87

 

SD

±0.98

SD

±0.97

SD

±0.45

 Samples of a loamy sand soil (0.7% org.C, pH (KCI) 5.6) were treated with the test item, amended with lucerne-grass-green meal (5 g/kg dry weight soil) and incubated in the dark at 20± 2 °C and 40% of the maximum water holding capacity.

Mean values (MV) and standard deviations (SD) from 3 soil samples (R)

Table 3: Rates of nitrogen transformation / day: individual values and mean values

Time lnterval (Days)

Test item / kg dry weight Soil

 

0 mg

0.07 mg

0.33 mg

 

mg Nitrogen (N) / kg dry weight Soil / Time lnterval

 

 

Nitrate-N

 

Nitrate-N

% Diff.

 

Nitrate-N

% Diff.

0-7

R1

-2.20

R4

-2.26

1

R7

-1.95

11

R2

-2.27

R5

-2.05

RB

-1.85

R3

-2.15

R6

-2.23

R9

-2.08

MV

-2.21

MV

-2.18

MV

-1.96

SD

±0.06

SD

±0.11

SD

±0.11

7-14

R1

0.74

R4

0.80

0

R7

0.82

4

R2

0.99

R5

0.74

RB

0.76

R3

0.72

R6

0.90

R9

0.97

MV

0.82

MV

0.81

MV

0.85

SD

±0.15

SD

±0.08

SD

±0.11

14-28

R1

1.03

R4

1.13

7

R7

0.97

0

R2

0.91

R5

1.00

RB

0.97

R3

1.04

R6

1.05

R9

1.03

MV

0.99

MV

1.06

MV

0.99

SD

±0.07

SD

±0.07

SD

±0.03

Samples of a loamy sand soil (0.7% org. C, pH (KCI) 5.6) were treated with the test item, amended with lucerne-grass-green meal (5 g/kg dry weight soil) and incubated in the dark at 20 ± 2 °C and 40% of the maximum water holding capacity.

(a) calculated from the individual and mean values of the respective intervals presented in Table 2.

% Diff. = % Difference (absolute values)

No significant difference between treated and untreated soil samples (t-test with 5 % probability of error).

Validity criteria fulfilled:
yes
Remarks:
The variation of nitrate concentrations between replicate control samples should be less than ± 15%, see Table 1 in "Any other information on results incl. tables".
Endpoint:
toxicity to soil microorganisms
Type of information:
experimental study
Adequacy of study:
key study
Study period:
23 Mar - 20 Apr 2004
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 217 (Soil Microorganisms: Carbon Transformation Test)
Version / remarks:
adopted 2000
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
Ministerium fur Umwelt, Raumordnung und Landwirtschaft des Landes Nordrhein-Westfalen, Germany
Analytical monitoring:
no
Vehicle:
no
Details on preparation and application of test substrate:
AMENDMENT OF SOIL
- Type of organic substrate: 2000 mg glucose/kg dry weight soil

APPLICATION OF TEST SUBSTANCE TO SOIL
- Method: Sieved soil (2 mm) was treated with either 10 g ground quartz sand/kg dry weight soil (control) or a mixture of quartz sand and test item (0.07 mg or 0.33 mg/kg dry weight soil). The samples were mixed in 3 liter polyethylene containers by rolling on a gyrowheel mixer for 15 min at 50 rpm. Since the soil was not extracted and analyzed, values given for the active ingredients are nominal. The test item concentrations 0.05 kg and 0.25 kg test item/ha corresponded to 0.07 mg and 0.33 mg test item/kg dry weight soil were used in the test.

VEHICLE: no vehicle was used
Test organisms (inoculum):
soil
Total exposure duration:
28 d
Test temperature:
20 ± 2 °C
Moisture:
40% of the maximum water holding capacity
Details on test conditions:
TEST SYSTEM
- Testing facility: Bayer CropScience AG
- Test container: 500 mL brown glass bottle and and were closed with parafilm
- Amount of soil: 350 g dry weight
- No. of replicates per concentration: 3
- No. of replicates per control: 3

SOIL INCUBATION
- Method: Three replicates were prepared per treatment. The quantity of product was added to the soil. The soil was held in the dark at 20 ± 2 °C and about 40% of the maximum water holding capacity (WHCmax).

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Geographical reference of sampling site (latitude, longitude): The soil used is from Field plot F on the Bayer AG's experimental farm Laacherhof, Germany located at a latitude of 51 °4' north and a longitude of 6°55' east.
- History of site: Plant protection chemicals have not been used on this field since 1981. The plot has been under grass and has not been treated with fertilizers since 1996. On March 07, 2000 the plot was plowed and then freshly planted with grass.
- Vegetation cover: grass
- Treatments with pesticides or fertilizers: no plant protection chemicals, no fertilizers has been used for the field
- Accidental contamination: no
- Depth of sampling: 5 cm
- Soil taxonomic classification: agricultural soil
- 2000 - 630 (µm): 4.0%
- 630 - 200 (µm): 52.5%
- 200 - 63 (µm): 21.6%
- 63 - 20 (µm): 6.9%
- 20 - 6.3 (µm): 4.3%
- 6.3 - 2.0 (µm): 4.4%
- 2.0 - 0 (µm): 6.3%
- Soil classification system: loamy sand
- pH (in water): 6.7
- Initial nitrate concentration for nitrogen transformation test (mg nitrate/kg dry weight): the soil held 12.58 mg nitrate-N/kg dry weight soil and 0.67 mg ammonium-N- and 12.58 mg nitrate-N/kg dry weight soil
- Cation exchange capacity: 6.4 meq/100 g dry weight soil
- Pretreatment of soil: Soil was passed through a 2 mm sieve and was treated with either 10 g ground quartz sand/kg dry weight soil (control) or a mixture of quartz sand test item (0.07 mg or 0.33 mg/kg dry weight soil). The samples were mixed in 3 liter polyethylene containers by rolling on a gyrowheel mixer for 15 min at 50 rpm.
- Storage (condition, duration): at 4 ± 2 °C
- Initial microbial biomass as % of total organic C: 0.7%

DETAILS OF PREINCUBATION OF SOIL (if any): no additional information

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Effects on non-target soil microorganisms on day 0, and after 7,14 and 28 days of incubation based to the glucosestimulatined soil respiration

VEHICLE CONTROL PERFORMED: no

RANGE-FINDING STUDY: no range-finding test was performed
Nominal and measured concentrations:
nominal concentrations: 0.07 mg test item/kg dry weight soil (0.05 kg test item/ha) and 0.33 mg test item/kg dry weight soil (0.25 kg test item/ha)
Reference substance (positive control):
yes
Remarks:
Sodium chloride
Duration:
28 d
Dose descriptor:
other: Difference to the control < 25%
Effect conc.:
0.07 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
respiration rate
Remarks on result:
other: test item had no influence on carbon transformation
Duration:
28 d
Dose descriptor:
other: Difference to the control < 25%
Effect conc.:
0.33 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
respiration rate
Remarks on result:
other: Test item had no influence on carbon transformation
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
>= 0.316 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
respiration rate
Remarks on result:
other: not part of the report but derivable from the reported results
Duration:
28 d
Dose descriptor:
EC10
Effect conc.:
> 0.316 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
respiration rate
Remarks on result:
other: not part of the report but derivable from the reported results
Details on results:
- Effect concentrations exceeding solubility of substance in test medium: During the 28-day tests, the recommended field rate of test item (0.05 kg test item/ha) and the 5-fold overdose of the test item had no influence on carbon transformation after addition of glucose to a loamy sand.
Results with reference substance (positive control):
- Results with reference substance valid? yes
- Relevant effect levels: In tests (non-GLP) with the agricultural soil described above, 16 g NaCI/kg dry weight soil had distinct and longterm (> 28 days) influences on microbial mineralization of carbon.
Reported statistics and error estimates:
All calculations were performed using Microsoft Excel 97. The % differences in the quantities of COa/h/kg dry weight soil formed between control soils and treated soils were expressed as absolute values and determined as follows: ((sum of treatment - sum of control)/sum of control) x 100 % = % difference. Homogeneity of variances was determined by F-test (significance level 5 %). Depending on the results of the F-Tests, the appropriate T-Tests were performed. In the T-Tests the sum of 12 hours of the mean values of CGVh/kg dry weight from control soils and treated soils were compared. The statistical calculations were carried out using Microsoft Excel 97 software according to the formulars of Sachs (Sachs, 1978).

During the 28-day tests, 0.07 mg test item/kg dry weight soil (0.05 kg test item/ha) and 0.33 mg test item/kg dry weight soil (0.25 kg test item/ha) had no influence on respiration after adding glucose to the loamy sand soil. After an exposure of 28 days the difference in the carbon dioxide production does not exceed the trigger value of 25% (mentioned in the guidelines). In this study, the highest coefficient of variation (CV) between carbon dioxide concentration in replicate control samples was 2.3% (14 days after treatment) and thus did not exceed the recommended limit <15%.

Table 1: Carbon transformation on day 0, just after treatment: Individual Values

Hours after addition of Glucose

mg Carbon Dioxide / Hour / kg dry weight Soil

Test Item/kg dry weight Soil

0 mg

Individual Values

0.07 mg

Individual Values

0.33 mg

Individual Values

1

14.17

14.53

13.57

13.83

13.50

14.00

13.00

13.33

14.33

2

14.77

14.83

14.00

13.73

13.33

13.97

14.00

14.17

13.83

3

14.77

15.60

14.77

14.30

14.27

14.10

14.10

14.10

14.10

4

15.33

15.33

14.67

13.83

14.33

13.83

14.07

14.47

14.67

5

15.50

16.00

15.50

14.83

15.07

14.50

14.67

15.50

14.83

6

16.83

18.33

16.67

15.67

15.83

15.83

15.83

16.17

16.17

7

18.67

18.50

18.67

17.00

17.43

17.50

17.00

17.17

17.93

8

20.33

20.67

20.50

18.83

19.17

19.17

18.50

20.17

20.17

9

23.00

23.00

23.17

20.83

21.83

20.83

20.33

22.00

21.70

10

26.07

26.50

25.63

22.50

23.83

22.67

22.50

24.17

24.67

11

28.33

30.00

29.17

25.00

27.27

26.23

25.50

28.00

28.83

12

35.33

34.67

34.50

29.43

31.50

30.60

29.67

32.70

32.00

Sum

243.10

247.97

240.80

219.80

227.37

223.23

219.17

231.93

233.23

MV of Sum

243.96

223.47*

228.11

SD

3.7

3.8

7.8

% CV

1.5

 

 

% of control

100

92

94

% Difference

-

8

6

Samples of a loamy sand soil (0.7% org. C, pH (KCI) 5.6) were treated with the test item., then immediately amended with 2000 mg glucose/kg dry weight soil and placed on an automated respirometer (gas analyzer).

Mean values (MV) and standard deviations (SD) from 3 soil samples.

% CV = Coefficient of Variation: (SD/MV of sum) x 100%.

% Difference (absolute value) = ((MV of sum of treatment- MV of sum of control)/MV of sum of control) x 100%.

* Significant difference between treated and untreated soil samples (t-test with 5 % probability of error).

Table 2: Carbon transformation 14 days after treatment: Individual Values

Hours after addition of Glucose

mg Carbon Dioxide / Hour / kg dry weight Soil

Test Item/kg dry weight Soil

0 mg

Individual Values

0.07 mg

Individual Values

0.33 mg

Individual Values

1

12.17

12.50

12.87

12.00

11.50

11.67

11.33

11.50

11.83

2

12.83

14.00

13.00

12.50

12.50

12.67

12.33

13.00

12.83

3

12.47

12.33

12.00

12.33

11.83

11.83

11.17

11.67

12.17

4

12.47

13.47

12.97

12.70

12.13

11.63

11.80

12.47

12.97

5

13.17

13.27

13.33

13.17

12.83

13.00

12.33

13.33

13.00

6

13.50

14.23

14.00

13.13

12.83

13.17

13.50

13.67

13.67

7

14.67

15.50

14.67

13.97

13.73

13.33

13.67

14.17

14.33

8

15.67

16.87

16.23

15.00

14.17

14.17

13.83

14.67

15.50

9

17.17

18.17

17.50

15.83

15.67

15.10

15.67

17.00

17.50

10

19.00

20.50

19.83

17.33

17.17

17.50

17.77

18.50

19.17

11

21.50

23.17

23.33

19.33

17.83

18.17

19.40

20.50

21.83

12

25.53

25.20

25.20

21.57

20.70

20.37

22.20

22.87

24.70

Sum

190.13

199.20

194.93

178.87

172.90

172.60

175.00

183.33

189.50

MV of Sum

194.76

174.79*

182.61

SD

4.5

3.5

7.3

% CV

2.3

 

 

% of control

100

90

94

% Difference

-

10

6

Samples of a loamy sand soil (0.7% org.C, pH (KCI) 5.6) were treated with test item, incubated in the dark at 20 ± 2 °C and 40% of the maximum water holding capacity for 14 days, then amended with 2000 mg glucose/kg dry weight soil and placed on an automated respirometer (gas analyzer).

Mean values (MV) and standard deviations (SD) from 3 soil samples.

% CV = Coefficient of Variation: (SD/MV of sum) x 100%.

% Difference (absolute value) = ((MV of sum of treatment- MV of sum of control)/MV of sum of control) x 100%.

* Significant difference between treated and untreated soil samples (t-test with 5 % probability of error).

Table 3: Carbon transformation 28 days after treatment: Individual Values

Hours after addition of Glucose

mg Carbon Dioxide / Hour / kg dry weight Soil

Test Item./kg dry weight Soil

0 mg

Individual Values

0.07 mg

Individual Values

0.33 mg

Individual Values

1

12.17

12.83

12.17

11.67

11.50

11.83

11.00

12.00

11.83

2

12.17

12.93

13.27

12.67

13.17

12.67

13.17

13.00

12.67

3

13.00

12.50

12.70

12.00

11.83

11.17

12.00

11.93

11.50

4

13.17

13.17

12.27

12.33

12.17

12.33

12.33

12.00

12.67

5

13.67

12.83

12.47

12.17

11.87

12.33

11.83

11.67

12.33

6

13.33

13.00

13.33

12.67

12.33

12.83

13.00

12.83

12.83

7

14.33

13.67

14.00

13.33

13.17

13.17

13.33

13.33

13.50

8

15.50

14.67

15.17

14.33

14.00

14.13

14.00

14.50

14.80

9

15.67

15.33

14.40

15.50

14.67

14.50

14.67

15.00

15.67

10

17.43

17.00

17.07

16.73

15.90

16.73

16.90

16.57

17.33

11

19.00

19.20

18.83

17.83

17.30

17.53

18.67

18.00

19.83

12

22.67

21.33

22.00

20.83

20.00

19.50

20.50

20.83

21.67

Sum

182.10

178.47

177.67

172.07

167.90

168.73

171.40

171.67

176.63

MV of Sum

179.41

169.57*

173.23

SD

2.4

2.2

2.9

% CV

1.3

 

 

% of control

100

95

97

% Difference

-

5

3

Samples of a loamy sand soil (0.7% org. C, pH (KCI) 5.6) were treated with test item, incubated in the dark at 20 ± 2 °C and 40% of the maximum water holding capacity for 28 days, then amended with 2000 mg glucose/kg dry weight soil and placed on an automated respirometer (gas analyzer).

Mean values (MV) and standard deviations (SO) from 3 soil samples.

% CV = Coefficient of Variation: (SO/MV of sum) x 100%.

% Oifference (absolute value) = ((MV of sum of treatment- MV of sum of control)/MV of sum of control) x 100%.

* Significant difference between treated and untreated soil samples (t-test with 5% probability of error).

 

Validity criteria fulfilled:
yes
Remarks:
The variation between replicate control samples should be less than ± 15%, see Table 1 in "Any other information on results incl. tables".

Description of key information

NOEC (28 d) ≥0.316 mg a.s./kg soil (OECD 216)
NOEC (28 d) ≥0.316 mg a.s./kg soil (OECD 217)

Key value for chemical safety assessment

Long-term EC10 or NOEC for soil microorganisms:
0.316 mg/kg soil dw

Additional information

Two experimental studies are available investigating the potential effects of the test item (CAS 874195-61-6) on nitrogen (M-080546-01-2) and carbon transformation (M-080753-01-2) of terrestrial microorganisms.The studies were performed according to OECD guideline 216 and 217 under GLP conditions.For both studies a loamy sand soil was exposed for 28 d to concentrations of 0.07 mg and 0.33 mg test item/kg dry weight soil (application rates were equivalent to 0.05 kg and 0.25 kg test item/ha).
In case of nitrogen transformation, Lucerne-grass-green meal was added to the soil (5 g/kg dry weight soil) to stimulate nitrogen transformation. The substance caused no adverse effects (OECD 216) on the soil nitrogen transformation (expressed as NO3-N-production) at the end of the 28 d incubation period. The study was performed in a field soil at concentrations up to 0.316 mg a.s./kg soil dry weight. The NOEC (28 d) was therefore reported as ≥ 0.316 mg a.s./kg soil. In case of carbon transformation glucose was added to the soil samples (2 g/kg dry weight soil) to induce maximum respiration rate. During the 28-day test, no influence on carbon transformation after addition of glucose to a loamy sand was recorded. The NOEC (28 d) was therefore reported as ≥0.316 mg a.s./kg soil.