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Ecotoxicological information

Toxicity to terrestrial plants

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Endpoint:
toxicity to terrestrial plants: long-term
Type of information:
experimental study
Adequacy of study:
key study
Study period:
27 April 2004 to 26 August 2004
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 208 (Terrestrial Plants Test: Seedling Emergence and Seedling Growth Test)
Version / remarks:
Proposal for Updating Guideline 208, Draft Document July 2000
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
6.75% solution of 2,6-dichlorobenzonitrile
Analytical monitoring:
no
Details on sampling:
Additional pots were treated with 3200 g test material/ha for analytical verification.
Vehicle:
no
Details on preparation and application of test substrate:
- Method of application to soil surface (if used): to ensure a homogenous distribution, the test material was mixed with a fixed amount of sand. After grinding the sand to quartz powder, the granules were added and ground until the sand-granule mixture was homogenous. This mixture was then spread homogeneously onto the moist soil.
Species:
Allium cepa
Plant group:
Monocotyledonae (monocots)
Details on test organisms:
- Common name: onion
- Plant family: Liliaceae
Species:
Avena sativa
Plant group:
Monocotyledonae (monocots)
Details on test organisms:
- Common name: oat
- Plant family: Poaceae
Species:
Helianthus annuus
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Common name: sunflower
- Plant family: Compositeae
Species:
Lycopersicon esculentum
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Common name: tomato
- Plant family: Solanaceae
Species:
Pisum sativum
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Common name: pea
- Plant family: Fabaceae
Species:
Raphanus sativus
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Common name: radish
- Plant family: Brassicaceae
Test type:
other: seedling emergence and seedling growth
Study type:
laboratory study
Substrate type:
natural soil
Limit test:
no
Total exposure duration:
21 d
Details on test conditions:
TEST SYSTEM
- Testing facility: growth chamber
- Test container (type, material, size): commercial plastic flower pots - 14 cm diameter
- Method of seeding: seeds were manually introduced
- No. of seeds per container: 5
- No. of replicates per treatment group: 8
- No. of replicates per control: 8
- No. of replicates per vehicle control: 8

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Soil type: sandy loam
- Organic carbon (%): 1.02 ± 0.17 %

GROWTH CONDITIONS
- Photoperiod: 16 hours light/8 hours dark
- Light intensity and quality: 5320-17690 lux
- Day/night temperatures: 22.51-26.30 ºC (day); 16.68-18.61 ºC (night)
- Relative humidity (%): 45.90-69.90 (day); 44.20-81.80 (night)
- Watering regime and schedules: daily when necessary via bottom watering through saucers

EFFECT PARAMETERS MEASURED:
- Fresh weight of all plants of a pot - 21 days after application
- Mortality - 21 days after application
- Germination - 7, 14 and 21 days after application
- Phytotoxicity rating system (if used): EPPO Standard PP 1/135 - 7, 14 and 21 days after application
- Growth stages - on application day and 21 days after application

TEST CONCENTRATIONS
- Range finding study results used to determine the conditions for the definitive study: yes
Nominal and measured concentrations:
13.11, 32.77, 81.92, 204.8, 512, 1280 and 3200 g test material/ha
Reference substance (positive control):
no
Key result
Species:
Avena sativa
Duration:
21 d
Dose descriptor:
ER50
Effect conc.:
100.31 g/ha
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Key result
Species:
Allium cepa
Duration:
21 d
Dose descriptor:
ER50
Effect conc.:
206.78 g/ha
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Key result
Species:
Lycopersicon esculentum
Duration:
21 d
Dose descriptor:
ER50
Effect conc.:
452.1 g/ha
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Key result
Species:
Raphanus sativus
Duration:
21 d
Dose descriptor:
ER50
Effect conc.:
659.78 g/ha
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Key result
Species:
Pisum sativum
Duration:
21 d
Dose descriptor:
ER50
Effect conc.:
1 249.53 g/ha
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Key result
Species:
Helianthus annuus
Duration:
21 d
Dose descriptor:
ER50
Effect conc.:
1 365.04 g/ha
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Details on results:
Results of fresh weight measurements, germination, mortality, phytotoxicity and plant growth are summarised in Table 2.

Fresh weights - The fresh weights were statistically significant reduced in A. sativa, A. cepa and L. esculentum at 81.92 g test material/ha and higher. For R. sativus, P. sativum and H. annuus the fresh weights were statistically significant reduced at 204.8, 1280 and 3200 g test material/ha, respectively.

Mortality - Mortality was observed in A. sativa at dose rate of 81.92 g test materiall/ha and higher (statistically significant at 204.8 g test material/ha) and for A. cepa statistically significant mortality was observed at 1280 g test material/ha (21 %). For R. sativus and H. annuus mortality was observed at 1280 g test material/ha (3 %, each) but not at the higher rate 3200 g test material/ha. No mortality was observed in L. esculentum and P. sativum.

Germination - The germination rate was significantly reduced in A. sativa at a dose rate of 204.8 g test material/ha and higher, for A. cepa at 1280 g dichlobenil/ha, for R. sativus at 512 g test material/ha and higher, for L. esculentum at 1280 g test material/ha and higher and for P. sativum at 1280 g test material/ha and higher. The germination rate of H. annuus was not affected.

Phytotoxicity - Phytotoxic effects, observed on all plant species, was mainly observed as growth reduction. In addition, chlorosis were observed in R. sativus and P. sativum and necrosis occurred in P. sativum and H. annuus.

The effective (dose) rates (based on fresh weight measurements are summarised in Table 2.

Table 2: Summarised resaults of fresh weight, germination, mortality, phytotoxicity and plant growth

  Treatment (g/ha) Germination (%) Mortality (%) Fresh wt (g) Effect* (%) Phytotoxicity (%) Growth stage (BBCH)
 Species        21DAA   7DAA 14DAA 21DAA  
A. sativa Control  95  0  8.32  0  0  0  0  13
   13.11  93  0  8.09  -2.73  0  0  1  13
   32.77  98  0  8.19  -1.64  0  1  2  13
   81.92  90  3  5.11  -38.57  0  5  17  13
   204.8  53  14  1.17  -85.93  0  9  46  12 - 13
   512  0  -  -  -  -  -  -  -
A. cepa  Control  98  0  1.13  0  0  0  0  12
   32.77  88  0  0.90  -21.02  0  0  0  12
   81.92  90  0  0.70  -37.98  0  0  0  12
   204.8  95  0  0.69  -38.85  0  0  4  12
   512  83  0  0.45  -60.69  0  2  11  11 - 12
   1280  60  21  0.03  -97.53  0  12  60  012 - 11
L. esculentum  Control  95  0  8.45  0  0  0  0  13
   81.92  98  0  8.00  -5.32  0  0  0  13
   204.8  100  0  5.52  -34.59  0  0  0  13
   512  95  0  4.63  -45.19  0  0  3  13
   1280  60  0  1.32  -84.39  0  5  28  12
   3200  0  -  -  -  -  -  -  -
R. sativus  Control  95  0  13.00  0  0  0  0  14
   81.92  98  0  11.77  -9.42  0  0  0  14
   204.8  93  0  11.18  -13.95  0  0  0  14
   512  75  0  8.75  -32.70  0  7  9  14
   1280  73  0  2.73  -79.02  0  20  39  11 - 14
   3200  35  0  0.13  -98.97  0  30  78  09 - 12
P. sativum  Control  100  0  13.25  0  0  0  0  16
   81.92  98  0  13.84  4.47  0  0  0  16
   204.8  98  0  12.51  -5.55  0  0  0  16
   512  95  0  11.75  -11.31  0  0  0  16
   1280  70  0  6.93  -47.72  0  4  13  15
   3200  10  0  0.39  -97.06  0  22  67  10 - 14
H. annuus  Control  98  0  19.20  0  0  0  0  14
   81.92  100  0  18.84  -1.88  0  0  0  14
   204.8  100  0  18.90  -1.58  0  0  0  14
   512  98  0  17.76  -7.48  0  3  0  14
   1280  100  0  10.33  -46.19  0  29  24  14
   3200  83  0  2.01  -89.52  0  50  61  14

DAA = days after treatment

*negative vales mean the reduction compared to the control

Table 2: Effective Dose Rates

 Species  NOER (g/ha)  LOER (g/ha)  ER25 (g/ha)  ER50 (g/ha)
 A. sativa  32.77  81.92  65.31  100.31
 A. cepa  32.77  81.92  57.65  206.78
 L. esculentum  81.92  204.8  192.55  452.10
 R. sativus  81.92  204.8  353.50  659.78
 P. sativum  512  1280  802.31  1249.53
 H. annuus  512  1280  863.40  1365.04
Validity criteria fulfilled:
yes
Conclusions:
Under the conditions of the test, the most sensitive species, based on terms of fresh weight reduction, is Avena sativa with an ER50 value of 100.31 g test material/ha.
Executive summary:

In a GLP compliant seedling emergence and seedling growth test conducted in line with standardised guideline OECD 208, the effect of the test material on seedling emergence and seedling growth over 21 days was determined. The effect of the test material on seedling emergence and seedling growth in six species was tested at nominal concentrations of 13.11, 32.77, 81.92, 204.8, 512, 1280 and 3200 g test material/ha. Under the conditions of the test, the most sensitive species, based on terms of fresh weight reduction, is Avena sativa with an ER50 value of 100.31 g test material/ha.

Endpoint:
toxicity to terrestrial plants: long-term
Type of information:
experimental study
Adequacy of study:
key study
Study period:
19 April 2004 to 26 August 2004
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 208 (Terrestrial Plants Test: Seedling Emergence and Seedling Growth Test)
Version / remarks:
Proposal for Updating Guideline 208, Draft Document July 2000
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
6.75% solution of 2,6-dichlorobenzonitrile
Analytical monitoring:
no
Details on sampling:
One additional pot was treated with 8100 g test material/ha for analytical verification.
Vehicle:
no
Details on preparation and application of test substrate:
- Method of application to soil surface (if used): to ensure a homogenous distribution, the test material was mixed with a fixed amount of sand. After grinding the sand to quartz powder, the granules were added and ground until the sand-granule mixture was homogenous. This mixture was then spread homogeneously onto the moist soil.
Species:
Allium cepa
Plant group:
Monocotyledonae (monocots)
Details on test organisms:
- Common name: onion
- Plant family: Liliaceae
Species:
Avena sativa
Plant group:
Monocotyledonae (monocots)
Details on test organisms:
- Common name: oat
- Plant family: Poaceae
Species:
Helianthus annuus
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Common name: sunflower
- Plant family: Compositeae
Species:
Lycopersicon esculentum
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Common name: tomato
- Plant family: Solanaceae
Species:
Pisum sativum
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Common name: pea
- Plant family: Fabaceae
Species:
Raphanus sativus
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Common name: radish
- Plant family: Brassicaceae
Test type:
vegetative vigour test
Study type:
laboratory study
Substrate type:
natural soil
Limit test:
no
Total exposure duration:
21 d
Details on test conditions:
TEST SYSTEM
- Testing facility: growth chamber
- Test container (type, material, size): commercial plastic flower pots - 14 cm diameter
- Method of seeding: seeds were manually introduced
- No. of seeds per container: 5 (A. cepa and A.sativa) or 3 (all other species)
- No. of replicates per treatment group: 8 (A. cepa and A.sativa) or 14 (all other species)
- No. of replicates per control: 8 (A. cepa and A.sativa) or 14 (all other species)

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Soil type: sandy loam
- Organic carbon (%): 1.02 ± 0.17 %

GROWTH CONDITIONS
- Photoperiod: 16 hours light/8 hours dark
- Light intensity and quality: 5320-19240 lux
- Day/night temperatures: 22.51-25.34 ºC (day); 16.92-18.61 ºC (night)
- Relative humidity (%): 48.10-69.90 (day); 48.70-81.80 (night)
- Watering regime and schedules: daily when necessary via bottom watering through saucers

EFFECT PARAMETERS MEASURED:
- Fresh weight of all plants of a pot - 21 days after application
- Mortality - 21 days after application
- Phytotoxicity rating system (if used): EPPO Standard PP 1/135 - 7, 14 and 21 days after application
- Growth stages - on application day and 21 days after application

TEST CONCENTRATIONS
- Spacing factor for test concentrations: Monocots - 4; Dicots - 3
- Range finding study
- Results used to determine the conditions for the definitive study: yes
Nominal and measured concentrations:
Dicots: 100, 300, 900, 2700, 8100, 24300 and 72900 g test material/ha

Monocots: 0.49, 1.98, 7.91, 32, 127, 506 and 2025 g test material/ha
Reference substance (positive control):
no
Key result
Species:
Allium cepa
Duration:
21 d
Dose descriptor:
ER50
Effect conc.:
821.07 g/ha
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Key result
Species:
Lycopersicon esculentum
Duration:
21 d
Dose descriptor:
ER50
Effect conc.:
3 914.05 g/ha
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Key result
Species:
Avena sativa
Duration:
21 d
Dose descriptor:
ER50
Effect conc.:
4 406.52 g/ha
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Key result
Species:
Pisum sativum
Duration:
21 d
Dose descriptor:
ER50
Effect conc.:
32 308.33 g/ha
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Key result
Species:
Raphanus sativus
Duration:
21 d
Dose descriptor:
ER50
Effect conc.:
258 027.45 g/ha
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Key result
Species:
Helianthus annuus
Duration:
21 d
Dose descriptor:
ER50
Effect conc.:
136 379 g/ha
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Details on results:
Results of fresh weight measurements, mortality, phytotoxicity and plant growth are summarised in Table 2.

Fresh weights - A statistically significant reduction in fresh weights was observed at and above 127 g test material/ha for A. cepa at and above 900 g test material/ha onwards for L. esculentum, at and above 32 g test material/ha for A. sativa, 100 g test material/ha for P. sativum, 2700 g test material/ha for R. sativus and at and above 300 g test material/ha for H. annuus.

Mortality - Mortality was observed for A. cepa at 506 and 2025 g test material/ha (13 and 73 %). Mortality of A. cepa was statistically significant different from the controls at 2025 g as/ha. For all other plant species tested no mortality was observed.

Phytotoxicity - Phytotoxic effects were observed in all plant species and were mainly evident as growth reduction and chlorosis. In addition, necrosis was observed in L. esculentum, R. sativus and H. annuus.

The Effective Rates (based on fresh weights) are summarised in Table 2.

Table 2: Summarised results of fresh weight, germination, mortality, phytotoxicity and plant growth

  Treatment group (g/ha) Mortality (%) Fresh wt (g) Effect* (%) Phytotoxicity (%) Growth stage (BBCH)
 Species      21DAA    7DAA  14DAA  21DAA  At application  21DDA
A. cepa  Control  0  14.63  0  0  0  0  12  15
   1.98  0  12.25  -16.25  0  0  0    15
   7.91  0  12.35  -15.59  0  0  0    15
   32  0  12.61  -13.79  0  0  0    15
   127  0  12.08  -17.46  1  1  0    15
   506  13  10.87  -25.67  3  16  17    14 - 15
   2025  73  2.39  -83.66  11  76  80    14
L. esculentum  Control  0  40.82  0  0  0  0  14  17
   100  0  41.10  0.69  0  0  0    17
   300  0  40.70  -0.28  0  0  0    17
   900  0  34.93  -14.43  0  3  2    17
   2700  0  27.30  -33.12  6  14  18    16 - 17
   8100  0  9.69  -76.25  25  45  74    15 - 16
A. sativa  Control  0  31.04  0  0  0  0  13  16
   0.49  0  30.27  -2.50  0  0  0    16
   1.98  0  32.24  3.85  0  0  0    16
   7.91  0  30.05  -3.22  0  0  0    16
   32  0  25.32  -18.42  0  0  1    16
   127  0  25.10  -19.13  0  3  2    16
P. sativum  Control  0  17.38  0  0  0  0  14  70
   100  0  14.80  -14.84  0  0  0    70
   300  0  14.22  -18.20  0  0  0    70
   900  0  12.84  -26.10  0  0  0    70
   2700  0  17.09  -1.67  0  0  1    70
   8100  0  14.59  -16.07  0  5  3    67
   24300  0  9.55  -45.04  0  12  31    67
   72900  0  5.15  -70.35  0  28  53    39 - 63
R. sativus  Control  0  11.35    0  0  0  13 - 14  15
   100  0  11.33  -0.16  0  0  0    15
   300  0  10.40  -8.33  0  0  0    15
   900  0  10.94  -3.63  0  0  0    15
   2700  0  8.96  -21.03  4  9  10    14 - 15
   8100  0  9.35  -17.61  1  19  40    14 - 15
H. annuus  Control  0  40.83    0  0  0  14  19
   100  0  39.18  -4.04  0  0  0    19
   300  0  37.60  -7.89  0  0  0    19
   900  0  36.25  -11.21  0  3  0    18 - 19
   2700  0  33.86  -17.06  0  3  1    18 - 19
   8100  0  30.52  -25.25  7  18  24    18 - 19

DAA = days after treatment

*negative vales mean the reduction compared to the control

Table 2: Effective Dose Rates

 Species  NOER (g/ha)  LOER (g/ha)  ER25 (g/ha)  ER50 (g/ha)
 A. cepa  32  127  381.68  821.07
 L. esculentum  300  900  1827.46  3914.05
 A. sativa  7.9  32  229.49  4406.52
 P. sativum  <100  100  8486.94  32,308.33
 R. sativus  ≥8100  >8100  15,273.50  258,027.45
 H. annuus  100  300  8191.79  136,379.00
Validity criteria fulfilled:
yes
Conclusions:
Under the conditions of the test, the most sensitive species, based on terms of fresh weight reduction, is Allium cepa with an ER50 value of 821.07 g test material/ha. The ER50 values across all species ranged from 1 -258 kg test material/ha.
Executive summary:

In a GLP compliant vegetative vigour test conducted in line with standardised guideline OECD 208, the effect of the test material on vegetative vigour over 21 days was determined. The effect of the test material on vegetative vigour in six species was tested at nominal concentrations of 0.49, 1.98, 7.91, 32, 127, 506 and 2025 g/ha (monocots) and 100, 300, 900, 2700, 8100, 24300 and 72900 g/ha (dicots). Under the conditions of the test, the most sensitive species, based on terms of fresh weight reduction, is Allium cepa with an ER50 value of 821.07 g test material/ha.

Description of key information

The ER50 to the most sensitive species for seedling emergence, Avena sativa, was determined to be 100.31 g test material/ha according to a study performed in line with OECD Guideline 208.

The ER50 to the most sensitive species for vegetative vigour, Allium cepa, was determined to be 821.07 g test material/ha according to a study performed in line with OECD Guideline 208.

Key value for chemical safety assessment

Additional information

In a GLP compliant seedling emergence and seedling growth test conducted in line with standardised guideline OECD 208, the effect of the test material on seedling emergence and seedling growth over 21 days was determined. The effect of the test material on seedling emergence and seedling growth in six species was tested at nominal concentrations of 13.11, 32.77, 81.92, 204.8, 512, 1280 and 3200 g/ha. Under the conditions of the test, the most sensitive species, based on terms of fresh weight reduction, is Avena sativa with an ER50 value of 100.31 g test material/ha.

In a GLP compliant vegetative vigour test conducted in line with standardised guideline OECD 208, the effect of the test material on vegetative vigour over 21 days was determined. The effect of the test material on vegetative vigour in six species was tested at nominal concentrations of 0.49, 1.98, 7.91, 32, 127, 506 and 2025 g/ha (monocots) and 100, 300, 900, 2700, 8100, 24300 and 72900 g/ha (dicots). Under the conditions of the test, the most sensitive species, based on terms of fresh weight reduction, is Allium cepa with an ER50 value of 821.07 g test material/ha.