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Toxicity to terrestrial plants

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Endpoint:
toxicity to terrestrial plants: long-term
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Justification for type of information:
See the read-across report attached in Section 13.
Reason / purpose for cross-reference:
read-across source
Species:
Hordeum vulgare
Duration:
5 d
Dose descriptor:
NOEC
Effect conc.:
322 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
other: Mn2+
Basis for effect:
other: Root elongation
Endpoint:
toxicity to terrestrial plants: long-term
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Justification for type of information:
See the read-across report attached in Section 13.
Qualifier:
according to guideline
Guideline:
other: ISO 11269-1 (1993):barley root elongation
Principles of method if other than guideline:
The principle of this test is based on the growth of 6 pregerminated seeds per pot under controlled conditions (16h/8h cycle light/darkness, 20°C during night time, 70% humidity). for a period of 5 days. The endpoint of this assay is the mean length of the longest root of each seedling per pot.
GLP compliance:
yes
Remarks:
K.U.Leuven
Analytical monitoring:
yes
Details on sampling:
- Sampling method: Total metal concentrations in soil were determined after the post-spiking incubation time for both spiked and control soils.
Vehicle:
yes
Details on preparation and application of test substrate:
All soils were air dried at 25°C and sieved to <4mm. Soil samples were preincubated at a moisture content equivalent to 50% of pF2 for 7 days at 20°c.Uncontaminated soils were spiked with MnCl2 to seven concentrations (control plus six treatments; 0, 0.01, 0.032, 0.1, 0.32, 1 and 3.2 x CEC;). Added Mn concentrations in corresponding treatments differ between the 3 soils as their CEC values increase from 6 to 21 cmolc/kg. Additional deionised water was added together with the spike solution to adjust the soil moisture content to 75% of pF2.0. All soils were thoroughly mixed after amendments using laboratory spoons. These spiked and preincubated soils was used for all microbial assays.
Species:
Hordeum vulgare
Plant group:
Monocotyledonae (monocots)
Test type:
seed germination/root elongation toxicity test
Study type:
laboratory study
Substrate type:
natural soil
Total exposure duration:
5 d
Post exposure observation period:
Not reported
Test temperature:
20°C
pH:
Soil 1- 4.8
Soil 2- 6.3
Soil 3- 7.5
Moisture:
Deionised water was added together with the spike solution to adjust the soil moisture content to 75% of pF 2.0.
Details on test conditions:
TEST SYSTEM
- Test container : Cylinder pots, approximately 8 cm in diameter and 11 cm in height.
- Amount of soil: MnCl2 spiked and preincubated soil was divided in 500 g (dry weight basis) subsamples.
- Method of seeding: Seeds with a radical length lower than 2 mm were planted in each pot approximately 10 mm beneath the surface of the soil.
- No. of seeds per container: 6 seeds per pot
- No. of plants (retained after thinning): Not reported
- No. of replicates per treatment group: 3
- No. of replicates per control: 3


SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Geographic location: Soil 1: Kasterlee (Belgium), Soil 2: Ter Munck (Belgium), Soil 3: Nagyhorcsok (Hungary)
- Collection procedures: Soils were collected from uncontaminated topsoils with contrasting soil properties. Soils were collected with a metal spade from the plough layer.Stones and vegetation were cleared from the soil samples before being put in 60 litre plastic drums.
- Sampling depth (cm): Not reported
- Soil texture (if natural soil)
- % sand: Soil 1- 79%, Soil 2- 8%, Soil 3- 13%
- % silt: Soil 1- 9%, Soil 2- 70%, Soil 3- 54%
- % clay: Soil 1- 9%, Soil 2- 70%, Soil 3- 54%
- Organic carbon (%): Soil 1- 2.8 %, Soil 2- 0.9 %, Soil 3- 2.1 %
- Maximum water holding capacity (in % dry weight): Soil 1 - pF0 -47%, pF1.8 -31, pF2- 27. Soil 2- pF0 -43%, pF1.8 -34, pF2-34. Soil 3-pF0 -48%, pF1.8 -35, pF2- 34
- CEC(at pH soil cmolc/kg): Soil 1-6.3, Soil 2-12.2 , Soil 3-24.8
- Storage : Air dry, sieved soils were stored in 60 litre plastic drums.


GROWTH CONDITIONS
- Photoperiod: 16h/8h cycle light/darkness
- Light intensity and quality: Light intensity at canopy height was 650 µmol photons/ m2/ s
- Day/night temperatures: 20°C during light hours and 16°C during night time
- Relative humidity (%): 70 %

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
The longest root per seedling was measured after 5 days of growth.


TEST CONCENTRATIONS
- Results used to determine the conditions for the definitive study: The rationale behind this spiking procedure (Mn addition as a fraction of the CEC) was based on previous research on metal toxicity in the lab showing strong positive correlations between metal toxicity (ECx values) and soil CEC i.e. soils with high CEC generally have higher ECx values than soil with low CEC. Therefore it was postulated that higher Mn additions were required in soil with high CEC compared to soil with low CEC to have a similar ECx values.
Nominal and measured concentrations:
Soil 1- 0, 17, 55, 170, 545, 1703, 5450 - Added Mn (mg/kg)
Soil 2- 0, 34, 107, 335, 1072, 2251, 10724 -Added Mn (mg/kg)
Soil 3- 0, 68, 218, 681, 2180, 6813, 21800 -Added Mn (mg/kg)
See table 6
Reference substance (positive control):
no
Species:
Hordeum vulgare
Duration:
5 d
Dose descriptor:
NOEC
Effect conc.:
322 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
other: Mn2+
Basis for effect:
other: Root elongation
Details on results:
Not reported
Results with reference substance (positive control):
Not reported
Reported statistics and error estimates:
Dose-response curves were fitted by a log-logistic model (Doelman and Haanstra, 1989) using the Maquardt Method (proc NLIN, SAS® 9.1; Cary, NC, USA):

Y = c/[1 + exp(b(x-a))]

With y = response variable, c= response in the control treatment, b= slope parameter, x= logarithm of the added Mn concentration and a = logarithm of the EC50 value. The model was adjusted to account for hormesis effects (Schabenberger et al., 1999; Van Ewijk and Hoekstra, 1993). The ‘dose’ in this model is the added Mn concentration (nominal concentration); with the dose in the control soil attributed a very small value (e.g. 0.001 mg Mn/kg). The EC50 values and their 95% confidence interval, expressed as added Mo, are predicted from the appropriate parameters of the curve. The EC50 values, expressed as ‘total Mn’, are calculated from the sum of background Mn concentration of a soil and corresponding EC50 (expressed as added Mn). No observed Effect Concentration (NOEC) are the highest Mn concentration in the soil at which no significant adverse effects on plant/ microbial assay were observed compared to the control soil (P> 0.05), and Lowest Observed Effect Concentrations (LOEC) are the lowest Mn concentration in the soil at which a significant adverse effect on plant/ microbial assay were observed compared to the control soil (P<0.05). Significant effects were determined by ANOVA (Dennett’s test). The threshold values (NOEC, LOEC and EC50) were determined in terms of soil Mn (mg/kg).

Table 8.Soil Mn toxicity thresholds for barley root elongation in freshly spiked soils. Thresholds are expressed as added + background Mn and the Confidence Intervals (CI) of EC10 and EC50 are given

 

Soil

Background Mn Conc. (mg/kg)

Root length control soil (cm)

NOEC (mg/kg)

EC10

(mg/kg)

95% CI

EC50 (mg/kg)

95% CI

1

152

14.3

322

497

369-701

965

767-1226

2

461

12.9

796

820

742-920

1239

1081-1438

3

722

9.4

940

1081

1003-1181

1500

1342-1699

Table 9.Root elongation expressed as lengths (cm) in Mn freshly spiked soil

 

Added Mn(mg/kg)

Soil 1

Root Length (cm)

0

14

10.5

13.8

14.2

13.5

14.8

0

14.2

14.5

16

15.5

15

13.5

0

14.5

16

13.4

15.4

13.2

15.1

17

11.8

11

11.5

12.5

14.3

13.2

17

10.4

11.5

12.4

11.8

12

12.5

17

15.2

12.4

14.2

13.5

10.2

1

55

14.5

13

13

12.5

13.7

13.5

55

13.8

12.5

12

12

12

11.5

55

13.5

15

13.5

11

13.5

13.5

170

11.5

13.5

11

15

11.5

12.4

170

12.8

13.2

16

13

14.5

13.5

170

14.8

14.5

16.4

15

14

14.5

545

12

11.2

11.4

12

12.1

12

545

11.7

12.4

11

11.

12.7

11.4

545

7.2

4.5

5

5

5.5

4.8

1703

1.5

1.7

1.8

2

3

1.5

1703

2

1.4

1.5

1.3

2

1.5

1703

2.4

2.3

1.1

1.8

1.2

1.5

5450

0.5

0.8

0.8

0.5

0.6

1

5450

1

1

0.5

0.4

0.3

0.4

5450

1.2

1

1

0.4

0.4

1

 

  10.Root elongation expressed as lengths (cm) in Mn freshly spiked soil.

  

Added Mn(mg/kg)

Soil 2

Root Length (cm)

0

9.5

13.7

13

11

14.5

13.5

0

11

13.2

11

14.5

11.5

15

0

14.5

13

13.2

12

14.2

13

34

15.7

12.7

11.5

14.5

14.8

15

34

10

13

11.2

11

12.2

14

34

14.7

15

13.2

14

11.5

14

107

11.4

12.2

12

11.8

10.8

11

107

12.5

13.8

14.4

13.5

13.4

 

107

13.7

12.8

11.5

11.4

13.5

16

335

11.8

12.7

11.8

12.7

11.2

12

335

12.7

11.1

12.7

11

12.4

11.7

335

12.2

12.5

11.7

10.5

 

8.5

1072

4.8

3.5

4.4

4.4

4.1

3.5

1072

3.8

3.1

4.5

4.5

4

3.5

1072

3

1.2

1.5

1.1

0.5

0.5

3351

1.5

1.7

1.8

2

3

1.5

3351

2

1.4

1.5

1.3

2

1.5

3351

2.4

2.3

1.1

1.8

1.2

1.5

10724

0.5

0.8

0.8

0.5

0.6

1

10724

1

1

0.5

0.4

0.3

0.4

10724

1.2

1

1

0.4

0.4

1

 

Table 11.Root elongation expressed as lengths (cm) in Mn freshly spiked soil.

 

 

Added Mn(mg/kg)

Soil 3

Root Length (cm)

0

9.2

8.7

11

10.1

9.2

9.8

0

8.8

9

9.5

10.5

9.5

6

0

9

9.8

10

10.2

10.2

8.7

68

12.5

11.8

9.2

11

11

11.8

68

10.4

11.8

11

11.5

12.2

10

68

12.4

10.5

11.2

10

11

9.8

218

12.2

9.8

10.5

9

11.1

10.7

218

 13.7

12.5

10.7

11.4

9.5

11

218

12

12.7

11.4

12.7

10.2

 

681

6.7

3.2

3.5

3.7

2.8

2.7

681

3.8

2.7

5

5.1

2.5

2.7

681

6

5.8

5.4

4.4

4

3.7

2180

2.3

3.8

2.5

3.2

3

3.1

2180

2.2

1.7

2.1

1.7

1.7

1.8

2180

3.2

2.5

3.5

3

2.7

3

6813

1.5

1.7

1

1.2

1.1

1.1

6813

1.4

1.4

1.5

1.3

1

1.1

6813

1.2

1.3

1.1

1.5

1.5

1.5

21800

0.4

0.5

0.7

0.3

0.5

0.8

21800

1

1

0.5

0.4

0.3

0.4

21800

1.1

0.7

0.5

0.4

0.4

0.7
Conclusions:
Under the conditions of the study, increasing Mn concentration significantly affected the barley root length in the three soils. The lowest NOEC was 322 mg Mn/kg.
Endpoint:
toxicity to terrestrial plants: long-term
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Justification for type of information:
See the read-across report attached in Section 13.
Reason / purpose for cross-reference:
read-across source
Species:
Hordeum vulgare
Duration:
23 d
Dose descriptor:
NOEC
Effect conc.:
697 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
other: Mn2+
Basis for effect:
growth
Species:
Lactuca sativa
Duration:
23 d
Dose descriptor:
NOEC
Effect conc.:
322 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
other: Mn2+
Basis for effect:
growth
Endpoint:
toxicity to terrestrial plants: long-term
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Justification for type of information:
See the read-across report attached in Section 13.
Qualifier:
according to guideline
Guideline:
other: ISO 11269-2 (2005): Plant growth assay (2 endpoints: barley growth and lettuce growth).
Principles of method if other than guideline:
This phototoxicity test was based on the emergence and early stages of growth of terrestrial plant species in the presence of various concentrations of toxic substances in soil. Seeds were planted in pots containing soil to which the test substance has been added. The emergence of the seeds and amass (dry or fresh basis) of the aboveground plant biomass were determined and compared to those of plants grown in uncontaminated soil. The validity criterion of this assay was the number of seedlings that emerge per pot and the endpoint was the shoot yield of the plants.
GLP compliance:
yes
Remarks:
K.U.Leuven
Analytical monitoring:
yes
Details on sampling:
- Sampling method: Total metal concentrations in soil were determined after the post-spiking incubation time for both spiked and control soils.
Vehicle:
yes
Details on preparation and application of test substrate:
All soils were air dried at 25°C and sieved to <4mm. Soil samples were preincubated at a moisture content equivalent to 50% of pF2 for 7 days at 20°c.Uncontaminated soils were spiked with MnCl2 to seven concentrations (control plus six treatments; 0, 0.01, 0.032, 0.1, 0.32, 1 and 3.2 x CEC;). Added Mn concentrations in corresponding treatments differ between the 3 soils as their CEC values increase from 6 to 21 cmolc/kg. Additional deionised water was added together with the spike solution to adjust the soil moisture content to 75% of pF2.0. All soils were thoroughly mixed after amendments using laboratory spoons. These spiked and preincubated soils was used for all microbial assays.
Species:
Hordeum vulgare
Plant group:
Monocotyledonae (monocots)
Species:
Lactuca sativa
Plant group:
Dicotyledonae (dicots)
Test type:
other: Emergence and early stages of growth
Study type:
laboratory study
Substrate type:
natural soil
Total exposure duration:
23 d
Test temperature:
Growing condition :20°C during light hours and 16°C during night time
The dry mass of the shoots was determined as well after oven drying at 70°C.
pH:
Not reported
Moisture:
Deionised water was added together with the spike solution to adjust the soil moisture content to 75% of pF 2.0.
Details on test conditions:
TEST SYSTEM
- Test container : Non porous plastic pots with a top internal diameter of 85 mm was used.
- Amount of soil: 450 g (dry weight)
- Method of seeding: Twenty uniform, undressed seeds of the selected species was planted in each pot.
- No. of seeds per container: 20
- No. of replicates per treatment group: 4
- No. of replicates per control: 4

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Geographic location: Soil 1: Kasterlee (Belgium), Soil 2: Ter Munck (Belgium), Soil 3: Nagyhorcsok (Hungary)
- Collection procedures: Soils were collected from uncontaminated topsoils with contrasting soil properties. Soils were collected with a metal spade from the plough layer.Stones and vegetation were cleared from the soil samples before being put in 60 litre plastic drums.
- Sampling depth (cm): Not reported
- Soil texture (if natural soil)
- % sand: Soil 1- 79%, Soil 2- 8%, Soil 3- 13%
- % silt: Soil 1- 9%, Soil 2- 70%, Soil 3- 54%
- % clay: Soil 1- 2%, Soil 2- 12%, Soil 3- 18%
- Organic carbon (%): Soil 1- 2.8 %, Soil 2- 0.9 %, Soil 3- 2.1 %
- Maximum water holding capacity (in % dry weight): Soil 1 - pF0 -47%, pF1.8 -31, pF2- 27. Soil 2- pF0 -43%, pF1.8 -34, pF2-34. Soil 3-pF0 -48%, pF1.8 -35, pF2- 34
- CEC: Soil 1-6.3, Soil 2-12.2 , Soil 3-24.8
- Pretreatment of soil: Not reported
- Storage (condition, duration): Air dry, sieved soils were stored in 60 litre plastic drums.


GROWTH CONDITIONS
- Photoperiod: 16h/8h cycle light/darkness
- Day/night temperatures: 20°C during light hours and 16°C during night time
- Relative humidity (%): 70 %
- Any pest control method/fertilization (if used): Prior to the palnt assay, the preincubated and MnCl2 spiked soil was fertilised with 50 mg P/kg soil as KH2PO4 and 100 mg N/Kg as KNO3 prior to the 7 day equilibration period.


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
After 2 days of growth, emergence was determined within each pot and seedlings were thinned to give a total of five evenly spaced representative specimens of the plants in the pots. After an additional 21 days of growth, shoot biomass above the soil surface was removed and the fresh weight was determined immediately after cutting. The dry mass of the shoots was determined as well after oven drying at 70°C for 16 h.


VEHICLE CONTROL PERFORMED: no

TEST CONCENTRATIONS
- Results used to determine the conditions for the definitive study:The rationale behind this spiking procedure (Mn addition as a fraction of the CEC) was based on previous research on metal toxicity in the lab showing strong positive correlations between metal toxicity (ECx values) and soil CEC i.e. soils with high CEC generally have higher ECx values than soil with low CEC. Therefore it was postulated that higher Mn additions were required in soil with high CEC compared to soil with low CEC to have a similar ECx values.
Nominal and measured concentrations:
Nominal Concentration
Soil 1- 0, 17, 55, 170, 545, 1703, 5450 mg/kg
Soil 2- 0, 34, 107, 335, 1072, 3351, 10724 mg/kg
Soil 3- 0, 68, 218, 681, 2180, 6813, 21800 mg/kg
Reference substance (positive control):
no
Species:
Hordeum vulgare
Duration:
23 d
Dose descriptor:
NOEC
Effect conc.:
697 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
other: Mn2+
Basis for effect:
growth
Species:
Lactuca sativa
Duration:
23 d
Dose descriptor:
NOEC
Effect conc.:
322 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
other: Mn2+
Basis for effect:
growth
Details on results:
Not reported
Results with reference substance (positive control):
Not reported
Reported statistics and error estimates:
Dose-response curves were fitted by a log-logistic model (Doelman and Haanstra, 1989) using the Maquardt Method (proc NLIN, SAS® 9.1; Cary, NC, USA):

Y = c/[1 + exp(b(x-a))]

With y = response variable, c= response in the control treatment, b= slope parameter, x= logarithm of the added Mn concentration and a = logarithm of the EC50 value. The model was adjusted to account for hormesis effects (Schabenberger et al., 1999; Van Ewijk and Hoekstra, 1993). The ‘dose’ in this model is the added Mn concentration (nominal concentration); with the dose in the control soil attributed a very small value (e.g. 0.001 mg Mn/kg). The EC50 values and their 95% confidence interval, expressed as added Mo, are predicted from the appropriate parameters of the curve. The EC50 values, expressed as ‘total Mn’, are calculated from the sum of background Mn concentration of a soil and corresponding EC50 (expressed as added Mn). No observed Effect Concentration (NOEC) are the highest Mn concentration in the soil at which no significant adverse effects on plant/ microbial assay were observed compared to the control soil (P> 0.05), and Lowest Observed Effect Concentrations (LOEC) are the lowest Mn concentration in the soil at which a significant adverse effect on plant/ microbial assay were observed compared to the control soil (P<0.05). Significant effects were determined by ANOVA (Dennett’s test). The threshold values (NOEC, LOEC and EC50) were determined in terms of soil Mn (mg/kg).

After 21 days of growth, barley biomass weight in the control soils varied between 0.23 -0.48g dry biomass/pot. Biomass weight of both plants decreased with increasing MnCl2 addition, and no growth was observed in the highest Mn treatment of all soils. Barley growth in the soil 2 showed a significant hormesis effect. Barley biomass weight were significantly lower Mn additions of 0.32x CEC for soils 2 and 3 and 1x CEC for soil 1 compared to the control soil, corresponding to a range of 1533 -2902 mg Mn/kg soil. Addition of Mn at concentrations corresponding to 1 and 3.2xCEC gives no or very little germination of barley seeds.

Lettuce biomass weight in the control soils varied between 0.13 -0.63 g dry biomass/pot, and was affected by increasing Mn doses in the 3 soils. Effects occurred at 0.3xCEC for soil 1, 0.3xCEC for soil1, 0.03xCEC for soil 2 and 0.1xCEC for soil 3 corresponding to 568 -697 mg Mn/kg soil. No growth was observed in the 1 and 3.2xCEC treatment of all soils.

Table 8.Soil Mn toxicity thresholds for barley growth in freshly spiked soils. Thresholds are expressed as added + background Mn and the Confidence Intervals (CI) of EC10 and EC50 are given.

 

Soil

Background Mn Conc. (mg/kg)

Barley dry weight control soil (g)

NOEC (mg/kg)

EC10

(mg/kg)

95% CI

EC50 (mg/kg)

95% CI

1

152

0.40

697

360

13-1552

717

379-1558

2

461

0.38

796

971

888-1069

1373

1202-1583

3

722

0.27

1403

1613

1200-2317

2456

2139-2843

  

Table 9. Soil Mn toxicity thresholds for lettuce growth in freshly spiked soils. Thresholds are expressed as added+ background Mn and the confidence Intervals (CI) of EC10 and EC50 are given.

 

Soil

Background Mn Conc. (mg/kg)

Barley dry weight control soil (g)

NOEC (mg/kg)

EC10

(mg/kg)

95% CI

EC50 (mg/kg)

95% CI

1

152

0.22

322

343

268-467

450

359-582

2

461

0.60

495

494

472-560

698

604-855

3

722

0.20

940

869

788-1051

1159

1026-1350

Table 10. Evaluation of higher plants development (barley) in Mn freshly spiked soils expressed as dry mass weight (g)- Soil 1

 

Added Mn (mg/kg)

Soil 1

Emergence (viable seeds)

Barley dry weight (g)

0

19

0.4603

0

19

0.2304

0

19

0.4774

0

20

0.4307

17

19

0.3009

17

19

0.295

17

20

0.2184

17

14

0.2842

55

20

0.2564

55

18

0.2788

55

20

0.183

55

19

0.3515

170

17

0.2065

170

17

0.2277

170

17

0.2385

170

17

0.2703

545

20

0.1596

545

18

0.174

545

20

0.2439

545

18

0.2346

1703

3

0.0437

1703

1

0.0526

1703

0

0

1703

0

0

5450

0

0

5450

0

0

5450

0

0

5450

0

0

 

Table 11. Evaluation of higher plants development (barley) in Mn freshly spiked soils expressed as dry mass weight (g)- Soil 2

 

Added Mn (mg/kg)

Soil 2

Emergence (viable seeds)

Barley dry weight (g)

0

20

0.4150

0

19

0.3042

0

20

0.4038

0

20

0.4020

34

17

0.4000

34

20

0.5230

34

19

0.4314

34

19

0.3220

107

19

0.5310

107

18

0.4952

107

18

0.4942

107

18

0.5782

335

17

0.4311

335

20

0.4350

335

17

0.4305

335

17

0.1740

1072

3

0

1072

0

0.1757

1072

2

0

1072

0

0

3351

0

0

3351

0

0

3351

0

0

3351

0

0

10724

0

0

10724

0

0

10724

0

0

10724

0

0

  

Table 12. Evaluation of higher plants development (barley) in Mn freshly spiked soils expressed as dry mass weight (g)- Soil 3

 

Added Mn (mg/kg)

Soil 3

Emergence (viable seeds)

Barley dry weight (g)

0

20

0.2695

0

20

0.2750

0

20

0.2832

0

17

0.2804

68

14

0.2535

68

19

0.2775

68

20

0.2717

68

20

0.2979

218

17

0.3127

218

17

0.3194

218

15

0.3939

218

19

0.3277

681

17

0.3095

681

20

0.2908

681

17

0.2927

681

19

0.2278

2180

1

0.0929

2180

15

0.0764

2180

17

0.0817

2180

4

0.1294

6813

0

0

6813

0

0

6813

0

0

6813

0

0

21800

0

0

21800

0

0

21800

0

0

21800

0

0

 

 Table 13.Evaluation of higher plants emergence (Barley and Lettuce) in Mn freshly spiked soils.

  

Soil

Nominal added salt conc (mmol/kg)

Emergence of barley in MnCl2 treatment

Emergence of lettuce in MnCl2 treatment

1

9.9

19

12

31.0

2

9

99.2

0

0

2

19.5

3

9

61.0

0

0

195.2

0

0

3

39.7

15

12

124.0

0

0

396.9

0

0

 

 

Table 14. Evaluation of higher plants development (lettuce) in Mn freshly spiked soils expressed as dry mass weight (g)- Soil 1

  

Added Mn (mg/kg)

Soil 1

Emergence (viable seeds)

Lettuce dry weight (g)

0

14

0.1965

0

14

0.2285

0

15

0.1868

0

15

0.2596

17

15

0.1416

17

7

0.2022

17

7

0.211

17

4

0.1569

55

8

0.1469

55

4

0.2283

55

8

0.1863

55

10

0.0975

170

6

0.2147

170

8

0.1332

170

8

0.1695

170

7

0.1288

545

12

0.0295

545

11

0.0121

545

10

0.0292

545

12

0.0247

1703

10

<0.01

1703

10

<0.01

1703

8

<0.01

1703

8

<0.01

5450

0

0

5450

0

0

5450

0

0

5450

0

0

 

Table 15. Evaluation of higher plants development (lettuce) in Mn freshly spiked soils expressed as dry mass weight (g)- Soil 2

 

Added Mn (mg/kg)

Soil 2

Emergence (viable seeds)

Lettuce dry weight (g)

0

18

0.6314

0

20

0.5040

0

20

0.6223

0

17

0.6261

34

20

0.5590

34

18

0.4761

34

20

0.5455

34

9

0.5558

107

10

0.4760

107

17

0.2388

107

19

0.3218

107

20

0.3638

335

18

0.4404

335

18

0.2940

335

14

0.3611

335

10

0.0408

1072

8

0.0070

1072

5

0.0211

1072

14

0.0400

1072

8

0

3351

0

0

3351

0

0

3351

0

0

3351

0

0

10724

0

0

10724

0

0

10724

0

0

10724

0

0

 

Table 16. Evaluation of higher plants development (lettuce) in Mn freshly spiked soils expressed as dry mass weight (g)- Soil 3 

  

Added Mn (mg/kg)

Soil 3

Emergence (viable seeds)

Barley dry weight (g)

0

16

0.2911

0

15

0.2650

0

15

0.1322

0

15

0.2212

68

14

0.2011

68

20

0.2173

68

17

0.3355

68

17

0.2090

218

12

0.2027

218

12

0.2299

218

17

0.1667

218

15

0.1661

681

8

0.1137

681

10

0.0200

681

8

0.0818

681

7

0.0384

2180

10

0.0280

2180

12

0.0277

2180

11

0.0288

2180

14

0

6813

0

0

6813

0

0

6813

0

0

6813

0

0

21800

0

0

21800

0

0

21800

0

0

21800

0

0
Validity criteria fulfilled:
yes
Remarks:
Emergence in the control soils varied between 14 and 20 out of 20 seeds for the barley and lettuce growth assay. As such the experimental setup was valid for all soils and seedlings were thinned to 5 healthy evenly distributed seedlings per pot.
Conclusions:
Barley and, especially, lettuce growth was sensitively affected by MnCl2 additions to soil, and toxic concentrations varied between soils. The lowest NOEC for Barley growth was 697 mg Mn/kg and for lettuce growth, 322 mg Mn/kg.

Description of key information

Del Carmen Hernandez Soriano et al. (2009) Read Across from MnCl2 (Barley Root)


Under the conditions of the study, increasing Mn concentration significantly affected the barley root length in the three soils. The lowest NOEC was 322 mg Mn/kg.


 


Del Carmen Hernandez Soriano et al. (2009a) Read Across from MnCl2 (Barley and Lettuce Growth)


Barley and, especially, lettuce growth was sensitively affected by MnCl2 additions to soil, and toxic concentrations varied between soils. The lowest NOEC for Barley growth was 697 mg Mn/kg and for lettuce growth, 322 mg Mn/kg.

Key value for chemical safety assessment

Long-term EC10, LC10 or NOEC for terrestrial plants:
322 mg/kg soil dw

Additional information

Del Carmen Hernandez Soriano et al. (2009) Read Across from MnCl2 (Barley Root)


Under the conditions of the study, increasing Mn concentration significantly affected the barley root length in the three soils. The lowest NOEC was 322 mg Mn/kg. The study was awarded a reliability score of 2 in accordance with the criteria set forth by Klimisch et al. (1997).


 


Del Carmen Hernandez Soriano et al. (2009a) Read Across from MnCl2 (Barley and Lettuce Growth)


Barley and, especially, lettuce growth was sensitively affected by MnCl2 additions to soil, and toxic concentrations varied between soils. The lowest NOEC for Barley growth was 697 mg Mn/kg and for lettuce growth, 322 mg Mn/kg. The study was awarded a reliability score of 2 in accordance with the criteria set forth by Klimisch et al. (1997).