Pyrasulfotole

Toxicity to terrestrial plants

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

toxicity to terrestrial plants: long-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

30 Apr – 28 Jun 2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

other: OECD (Draft Document): Guideline for the testing of chemicals, Proposal for updating Guideline 208 „Terrestrial (Non-Target) Plant Test: 208 A: Seedling Emergence and Seedling Growth Test. 208 B: Vegetative Vigour Test."

Version / remarks:

Part A, July 2000

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPP 123-1 (Seedling Emergence Tier II)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Hessisches Ministerium für Umwelt, ländlichen Raum und Verbraucherschutz, Wiesbaden, Germany

Analytical monitoring:

yes

Details on sampling:

- Concentrations: Samples of the highest application rate were collected at day 0 and analysed for the test item. The samples were diluted with acetonitrile / purest water 20/80 (v/v)

Vehicle:

no

Details on preparation and application of test substrate:

- Method of application of test item and control: The spray solution was applied once, at test initiation on the soil surface. The blank control spray solution was 100 L deionized water. The test item was dissolved in deionized water and was applied once with 100 L/ha using an spray chamber equipped with an overhead nozzle (Lechner LU 367-02), with nozzle height set at 50 cm above the sprayed surface. The spray chamber volume was calibrated by weighing the amount of water applied to a known surface area.
- Controls: yes

Species:

Lolium perenne

Plant group:

Monocotyledonae (monocots)

Details on test organisms:

- Common name: Perennial ryegrass
- Plant family: Poaceae
- Variety: Deutsches Weidegras
- Source of seed: commercial sources via Bayer CropScience GmbH, Horticulture, H872, 65926 Frankfurt am Main
- Prior seed treatment/sterilization: Seeds used on the study had not been treated with pesticides or repellents prior to test initiation
- Seed storage: Seeds were stored in plastic boxes in the refrigerator

Species:

Avena sativa

Plant group:

Monocotyledonae (monocots)

Details on test organisms:

- Common name: Oat
- Plant family: Poaceae
- Variety: Flämings Nova
- Source of seed: commercial sources via Bayer CropScience GmbH, Horticulture, H872, 65926 Frankfurt am Main
- Prior seed treatment/sterilization: Seeds used on the study had not been treated with pesticides or repellents prior to test initiation
- Seed storage: Seeds were stored in plastic boxes in the refrigerator

Species:

Zea mays

Plant group:

Monocotyledonae (monocots)

Details on test organisms:

- Common name: Corn
- Plant family: Poaceae
- Variety: Lorenzo
- Source of seed: commercial sources via Bayer CropScience GmbH, Horticulture, H872, 65926 Frankfurt am Main
- Prior seed treatment/sterilization: Seeds used on the study had not been treated with pesticides or repellents prior to test initiation
- Seed storage: Seeds were stored in plastic boxes in the refrigerator

Species:

Hordeum vulgare

Plant group:

Monocotyledonae (monocots)

Details on test organisms:

- Common name: Spring barley
- Plant family: Poaceae
- Variety: Baroness
- Source of seed: commercial sources via Bayer CropScience GmbH, Horticulture, H872, 65926 Frankfurt am Main
- Prior seed treatment/sterilization: Seeds used on the study had not been treated with pesticides or repellents prior to test initiation
- Seed storage: Seeds were stored in plastic boxes in the refrigerator

Species:

Lycopersicon esculentum

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

- Common name: Tomato
- Plant family: Solanaceae
- Variety: Balkonstar
- Source of seed: commercial sources via Bayer CropScience GmbH, Horticulture, H872, 65926 Frankfurt am Main
- Prior seed treatment/sterilization: Seeds used on the study had not been treated with pesticides or repellents prior to test initiation
- Seed storage: Seeds were stored in plastic boxes in the refrigerator

Species:

Helianthus annuus

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

- Common name: Sunflower
- Plant family: Asteraceae
- Variety: Big Smile
- Source of seed: commercial sources via Bayer CropScience GmbH, Horticulture, H872, 65926 Frankfurt am Main
- Prior seed treatment/sterilization: Seeds used on the study had not been treated with pesticides or repellents prior to test initiation
- Seed storage: Seeds were stored in plastic boxes in the refrigerator

Species:

Beta vulgaris

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

- Common name: Sugar beet
- Plant family: Chenopodiaceae
- Variety: Achat
- Source of seed: commercial sources via Bayer CropScience GmbH, Horticulture, H872, 65926 Frankfurt am Main
- Prior seed treatment/sterilization: Seeds used on the study had not been treated with pesticides or repellents prior to test initiation
- Seed storage: Seeds were stored in plastic boxes in the refrigerator

Species:

Glycine max (G. soja)

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

- Common name: Soybean
- Plant family: Fabaceae
- Variety: Trail
- Source of seed: commercial sources via Bayer CropScience GmbH, Horticulture, H872, 65926 Frankfurt am Main
- Prior seed treatment/sterilization: Seeds used on the study had not been treated with pesticides or repellents prior to test initiation
- Seed storage: Seeds were stored in plastic boxes in the refrigerator

Species:

Brassica napus

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

- Common name: Oilseed rape
- Plant family: Brassicaceae
- Variety: Liratop
- Source of seed: commercial sources via Bayer CropScience GmbH, Horticulture, H872, 65926 Frankfurt am Main
- Prior seed treatment/sterilization: Seeds used on the study had not been treated with pesticides or repellents prior to test initiation
- Seed storage: Seeds were stored in plastic boxes in the refrigerator

Species:

Cucumis sativus

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

- Common name: Cucumber
- Plant family: Curcubitaceae
- Variety: Delikatess
- Source of seed: commercial sources via Bayer CropScience GmbH, Horticulture, H872, 65926 Frankfurt am Main
- Prior seed treatment/sterilization: Seeds used on the study had not been treated with pesticides or repellents prior to test initiation
- Seed storage: Seeds were stored in plastic boxes in the refrigerator

Test type:

seedling emergence and seedling growth test

Study type:

laboratory study

Substrate type:

natural soil

Limit test:

no

Total exposure duration:

21 d

Test temperature:

23 ± 5 °C day, 18 ± 5 °C night (temperature control set)

pH:

7.4

Moisture:

Humidity: 80% day, 95% night (glasshouse control settings)

Details on test conditions:

TEST SYSTEM
- Testing facility: Glasshouse conditions Bayer CropScience GmbH; Frankfurt (Main), Germany
- Test container (type, material, size): Commercial plastic flower pots (10 cm diameter)
- Method of seeding: Seeds were introduced manually in the soil, covered by up to 5 mm of soil and top watered immediately to facilitate germination. The size of seed determined the sowing depth, i.e. 5 mm diameter seeds covered by 5 mm soil and 2 mm diameter seeds covered by 2 mm soil.
- No. of seeds per container: Each pot contained 5 seeds. In total 40 seeds per treatment group were tested.
- No. of replicates per treatment group: 8 pots per treatment group
- No. of replicates per control: 8 pots per treatment group

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Geographic location: Standard soil (silty loam) from Bayer CropScience GmbH, Global Biology Herbicides, Horticulture, H 872, Industriepark Hoechst, 65926 Frankfurt am Main; Germany
- Pesticide use history at the collection site: Locally supplied soil steam pasteurized before use (120 °C vapour for about 30 minutes)
- Collection procedures: soil was sieved to 2 mm. The soil was analysed separately without GLP in the laboratory of Development Ecotoxicology, Bayer CropScience AG, 40789 Monheim, Germany
- Soil texture (if natural soil)
- % sand: 14.2%
- % silt: 65.1%
- % clay: 20.7%
- Soil taxonomic classification: Silty loam
- Organic carbon (%): 1.19%

GROWTH CONDITIONS
- Photoperiod: 16 h light, 8 h dark (glasshouse control settings)
- Light source and intensity: Natural daylight supplemented by artificial lighting to provide the required photoperiod. Regulation of light intensity >10000 lux lamps turn off, >20000 lux shading closing
- Day/night temperatures: 23 ± 5°C day, 18 ± 5°C night (temperature control set)
- Relative humidity (%): 80% day, 95% night (glasshouse control settings)
- Watering regime and schedules: Watered as required at least once per day. Following the initial top watering to facilitate germination of the seed, bottom watering was performed with saucers standing below each pot. Water was given and retained within the saucer according to the need of the plants in order to have an optimal water supply for plant growth. This was checked daily.
- Water source/type: Local tap water. Pots contained in plastic saucers in which the water was added
- Volume applied: test item was dissolved in deionized water and was applied once with 100 L/ha using an spray chamber
- Interval of applications: once at test start
- Method of application: with an overhead nozzle (Lechner LU 367-02), with nozzle height set at 50 cm above the sprayed surface
- Any pest control method/fertilization (if used): 2.4 g/L of granular fertilizer (Blaukorn) was added to the soil prior to sowing

EFFECT PARAMETERS MEASURED: Emergence, survival, phytotoxicity, growth stage, plant dry weight and length 21 d after application (observations on day 7, 14 and 21)

- Phytotoxicity rating system (if used): 0 (no injury/effect), 20%(indicates slight effect), 40% (indicates moderate effect), 60% (indicates severe effect), 80% (Indicates total plant effect), 100% (plant death).

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.0

Nominal and measured concentrations:

Control, 0.453, 0.906, 1.77, 3.53, 7.07,14.2, 28.3, 56.6, and 113 g a.i. /ha (nominal application rate)
Control, 0.01, 0.02, 0.039, 0.078, 0.156, 0.313, 0.625, 1.25, and 2.5 kg product/ha (nominal test concentration)
Control, 2.5, 1.25, 0.625, 0.313, 0.156 and 0.078 kg product / ha (nominal test concentrations for corn and oat)
Control, 1.25, 0.625, 0.313, 0.156, 0.078 and 0.039 kg product / ha (nominal test concentrations for barley)
Control, 0.625, 0.313, 0.156, 0.078, 0.039, 0.020 and 0.010 kg product / ha (nominal test concentrations for cucumber, oilseed rape, soybean, sunflower, sugar beet, tomato and ryegrass)

Reference substance (positive control):

no

Species:

Lycopersicon esculentum

Duration:

21 d

Dose descriptor:

EC50

Effect conc.:

0.008 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: plant biomass

Remarks on result:

other: equivalent to 6.12 g a.i./ha

Species:

Lycopersicon esculentum

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

>= 0.002 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: Plant biomass

Remarks on result:

other: equivalent to 1.77 g a.i./ha

Species:

Brassica napus

Duration:

21 d

Dose descriptor:

EC50

Effect conc.:

0.013 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: plant shoot length

Remarks on result:

other: equivalent to 9.61 g a.i./ha

Species:

Lycopersicon esculentum

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

>= 0.001 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: plant shoot length

Remarks on result:

other: equivalent to 0.89 g a.i./ha

Species:

Beta vulgaris

Duration:

21 d

Dose descriptor:

EC50

Effect conc.:

0.015 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: plant survival

Remarks on result:

other: equivalent to 11.28 g a.i./ha

Species:

other: Brassica napus, Beta vulgaris, Lycopersicon esculentum

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

>= 0.009 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: plant survival

Remarks on result:

other: equivalent to 7.08 g a.i./ha

Species:

other: Cucumis sativus, Brassica napus, Lycopersicon esculentum, Lolium perenne, Helianthus annuus L., Glycine max, Beta vulgaris

Duration:

21 d

Dose descriptor:

EC50

Effect conc.:

> 0.038 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

seedling emergence

Remarks on result:

other: equivalent to 28.3 g a.i./ha

Species:

other: Cucumis sativus, Brassica napus, Lycopersicon esculentum, Lolium perenne, Helianthus annuus L., Glycine max, Beta vulgaris

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.038 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

seedling emergence

Remarks on result:

other: equivalent to 28.3 g a.i./ha

Details on results:

- Typical symptoms: chlorosis, necrosis, abnormal growth and growth suppression with 5 of the species in this study, i.e. oilseed rape, sugar beet, sunflower, tomato and corn, which exhibited some or all of these symptoms, the severity of which increased with dose rate tested.
- Biological and analytical results in details are summarized with the tables in section "Any other information on results incl tables"

Reported statistics and error estimates:

Statistical Analysis: Mortality and biomass (plant dry weight) were compared using the ToxRat software for statistical analysis (version 2.09).
Detrimental Effect Levels: EC25 and EC50 with the 95 percent confidence limits as well as the LOEC (Lowest Observed Effect Concentration) and NOEC (No Observed Effect Concentration)

Endpoints:

The endpoints were reported as kg test item/ha and g a.i./ha. Based on these reported values the endpoints were recalculated to mg/kg soil which were not part of the report but based on the reported data of g a.i./h and a density of 1.5 g/cm³ and a soil depth of 5 cm.

Analytical Results:

Measured concentrations were 92.5% - 96.5% of nominal.

Table 1: Validity criteria in the untreated control for the seedling emergence and growth test with the test item

	Validity criteria for germination (% of sown seeds that emerged)	Actual germination achieved	Survival (% of emerged)	Phytotoxicity in untreated controls in %
Validity criteria			>90%	0%
Cucumber	80%	95%	92.1	0
Oilseed rape	75%	80%	100	0
Soybean	75%	82.5%	90.9	0
Sugar beet	55%	97.5%	100	0
Sunflower	80%	100%	97.5	0
Tomato	75%	90%	100	0
Barley	80%	85%	100	0
Corn	85%	100%	100	0
Oat	80%	97.5%	100	0
Ryegrass	75%	87.5%	100	0

 

Table 2: The mean phytotoxicity values at day 21 for the seedling emergence and growth test with the test item

Day 21 phytotoxicity summary (mean % damage) Dicot species
kg product/ha	Cucumber	Oilseed rape	Soybean	Sugar beet	Sunflower	Tomato
0.03773	15	88	0	95	95	-
0.01888	6	85	0	71	35	60
0.00944	0	66	0	30	4	33
0.00472	0	4	0	0	0	46
0.00236	0	0	0	0	0	33
0.001187	0	0	0	4	0	16
0.0006	0	0	0	0	0	0
Control	0	0	0	0	0	0

 

Day 21 phytotoxicity summary (mean % damage) Monocot species
kg product/ha	Barley	Corn	Oat	Ryegrass
0.1507	NT	13	0	NT
0.07551	0	0	0	NT
0.03773	0	0	0	0
0.01888	0	0	0	0
0.00944	0	0	0	0
0.00472	0	0	0	NT
0.00236	0	NT	NT	0
0.001187	NT	NT	NT	0
0.00006	NT	NT	NT	0
Control	0	0	0	0

a Percent effect values were calculated in comparison to controls.

NT = concentration not tested

- 100% mortality

Table 3: Analytical Results

Sample no.	Desired value mg/L	Result mg/L	Result % of theory	CV (%)
Recovery	550.04	548.80	99.8	0.6
SE04/006-2-001	0	0	--	--
SE04/006-2-002	566.25	523.82	92.5	0.1

Dicotyledonous species

Cucumber (Cucumis sativus)

The foliar application of the test item applied at seven dose rates of 0.03773, 0.01888, 0.00944 0.00472, 0.00236, 0.001187, 0.0006 mg a.i./kg soil resulted in no significant effects of seedling emergence and survival and therefore the NOEC was the highest rate tested in this study and the LOEC > this rate. Analysis of shoot length after 21 days resulted in EC25 and EC50 endpoints of 0.03921 and >0.03773 mg a.i./kg soil, respectively. Significant adverse effects on shoot length were apparent at dose rates including and above 0.01888 mg a.i./kg soil with a NOEC of 0.00944 mg a.i./kg soil. Analysis of plant biomass (shoot dry weight) at day 21 resulted in EC25 and EC50 endpoints of 0.0226 and >0.03773 mg a.i./kg soil, respectively with significant adverse effects only at the highest rate of 0.03773 mg a.i./kg soil and a NOEC of 0.0189 mg a.i./kg soil. Slightly symptoms of phytotoxicity were apparent at the two highest dose rates; there were marginal effects of plant growth as determined by growth stage analysis at day 21 at doses of 0.00236 mg a.i./kg soil and above.

Oilseed rape (Brassica napus)

The foliar application of the test item applied at seven dose rates of 0.03773, 0.01888, 0.00944 0.00472, 0.00236, 0.001187, 0.0006 mg a.i./kg soil resulted in no significant effects of seedling emergence and therefore the NOEC was the highest rate tested and the LOEC > this rate. Analysis of survival after 21 days resulted in EC25 and EC50 endpoints of 0.0116 and 0.0264 mg a.i./kg soil, respectively. Significant adverse effects on survival were apparent at dose rates including and above 0.01888 mg a.i./kg soil with a NOEC of 0.00944 mg a.i./kg soil. Analysis of shoot length after 21 days resulted in EC25 and EC50 endpoints of 0.00556 and 0.012813 mg a.i./kg soil, respectively. Significant adverse effects on shoot length were apparent at dose rates including and above 0.00944 mg a.i./kg soil with a NOEC of 0.00472 mg a.i./kg soil. Analysis of plant biomass (shoot dry weight) at day 21 resulted in EC25 and EC50 endpoints of 0.0058 and 0.01027 mg a.i./kg soil, respectively with significant adverse effects at the rates of 0.00944 mg a.i./kg soil and above. Therefore, the NOEC was 0.00472 mg a.i./kg soil. Symptoms of phytotoxicity wer apparent at rates of 0.00472 mg a.i./kg soil and above and development of plant growth, as determined by growth stage analysis at day 21 was reduced including and above 0.00944 mg a.i./kg soil.

Soybean (Glycine max)

The foliar application of the test item applied at seven dose rates of 0.03773, 0.01888, 0.00944 0.00472, 0.00236, 0.001187, 0.0006 mg a.i./kg soil resulted in no significant effects of seedling emergence, survival, shoot length and plant biomass and therefore the NOEC was the highest rate tested of 0.03773 mg a.i./kg soil in this study and the LOEC >this rate. Symptoms of phytotoxicity were not apparent at any dose rate however, there were marginal effects of plant growth as determined by growth stage analysis at day 21 at doses of 0.01888 mg a.i./kg soil and above.

Sugar beet (Beta vulgaris)

The foliar application of the test item applied at seven dose rates of 0.03773, 0.01888, 0.00944 0.00472, 0.00236, 0.001187, 0.0006 mg a.i./kg soil resulted in no significant effects of seedling emergence and therefore the NOEC was the highest rate tested in this study and the LOEC > this rate. Analysis of survival after 21 days resulted in EC25 and EC50 endpoints of 0.00852 and 0.01504 mg a.i./kg soil, respectively. Significant adverse effects on survival were apparent at dose rates including and above 0.01888 mg a.i./kg soilwith a NOEC of 0.00944 mg a.i./kg soil. Analysis of shoot length after 21 days resulted in EC25 and EC50 endpoints of 0.009667 and 0.01745 mg a.i./kg soil, respectively. Significant adverse effects on shoot length were apparent at dose rates including and above 0.00944 mg a.i./kg soilwith a NOEC of 0.00472 mg a.i./kg soil. Analysis of plant biomass (shoot dry weight) at day 21 resulted in EC25 and EC50 endpoints of 0.00924 and 0.01873 mg a.i./kg soil, respectively with significant adverse effects only at the highest rate of 0.03773 mg a.i./kg soil. This results in a NOEC of 0.00472 mg a.i./kg soil. Severe symptoms of phytotoxicity were apparent at rates including and above 0.00944 mg a.i./kg soil. Development of plant growth, as determined by growth stage analysis at day 21 was reduced at 0.001187 mg a.i./kg soiland including and above 0.00944 kg product/ha.

 

Sunflower (Helianthus annuus)

The foliar application of the test item applied at seven dose rates of 0.03773, 0.01888, 0.00944 0.00472, 0.00236, 0.001187, 0.0006 mg a.i./kg soil resulted in no significant effects of seedling emergence and therefore the NOEC was the highest rate tested in this study and the LOEC > this rate. Analysis of survival after 21 days resulted in EC25 and EC50 endpoints of 0.04199 and >0.03773 mg a.i./kg soil, respectively. Significant adverse effects on survival were apparent at 0.03773 mg a.i./kg soil with a NOEC of 0.01888 mg a.i./kg soil. Analysis of shoot length after 21 days resulted in EC25 and EC50 endpoints of 0.0114 and 0.02108 mg a.i./kg soil, respectively. Significant adverse effects on shoot length were apparent at dose rates including and above 0.00472 mg a.i./kg soilwith a NOEC of 0.00236 mg a.i./kg soil. Analysis of plant biomass (shoot dry weight) at day 21 resulted in EC25 and EC50 endpoints of 0.01693 and 0.02907kg product/ha, respectively with significant adverse effects including and above 0.01888 mg a.i./kg soil with a NOEC of 0.00944 mg a.i./kg soil. Symptoms of phytotoxicity were apparent at rates including and above 0.00944 mg a.i./kg soil. Development of plant growth, as determined by growth stage analysis at day 21 was reduced at dose rates including and above 0.00236 mg a.i./kg soil.

Tomato (Lycopersicum esculentum)

The foliar application of the test item applied at seven dose rates of 0.03773, 0.01888, 0.00944 0.00472, 0.00236, 0.001187, 0.0006 mg a.i./kg soil resulted in no significant effects of seedling emergence and therefore the NOEC was the highest rate tested in this study and the LOEC > this rate. Analysis of survival after 21 days resulted in EC25 and EC50 endpoints of 0.01256 and 0.01957 mg a.i./kg soil, respectively. Significant adverse effects on survival were apparent at 0.01888 and 0.03773 mg a.i./kg soil with a NOEC of 0.00944 mg a.i./kg soil. Analysis of shoot length after 21 days resulted in EC25 and EC50 endpoints of 0.00496 and >0.03773 mg a.i./kg soil, respectively. Significant adverse effects on shoot length were apparent at dose rates including and above 0.00236 mg a.i./kg soil with a NOEC of 0.001187 mg a.i./kg soil. Analysis of plant biomass (shoot dry weight) at day 21 resulted in EC25 and EC50 endpoints of 0,001693333 and 0.00816 mg a.i./kg soil, respectively with significant adverse effects including and above 0.00472 mg a.i./kg soil with a NOEC of 0.00236 mg a.i./kg soil. Symptoms of phytotoxicity were apparent a rates including and above 0.001187 mg a.i./kg soil. Development of plant growth, as determined by growth stage analysis at day 21 was reduced at dose rates including and above 0.001187 mg a.i./kg soil.

 

Monocotyledonous species

Barley (Hordeum vulgare)

The foliar application of the test item applied at six dose rates of 0.07551, 0.03773, 0.01888, 0.00944, 0.00472 and 0.00236 mg a.i./kg soil resulted in no significant effects of seedling emergence, survival, shoot length and plant biomass and therefore the NOEC was the highest rate tested of 0.07551 mg a.i./kg soil in this study and the LOEC > this rate. Symptoms of phytotoxicity were not apparent at any dose rate and there was no adverse effect on plant growth.

Corn (Zea mays)

The foliar application of the test item applied at six dose rates of 0.1507, 0.07551, 0.03773, 0.01888, 0.00944 and 0.00472 mg a.i./kg soil resulted in no significant effects of seedling emergence, survival, shoot length and plant biomass and therefore the NOEC was the highest rate tested of 0.1507 mg a.i./kg soil in this study and the LOEC > this rate. Symptoms of phytotoxicity were apparent at the highest dose rate of 0.1507 mg a.i./kg soil and there was no adverse effect on plant growth

Oats (Avena sativa)

The foliar application of the test item applied at six dose rates of 0.1507, 0.07551, 0.03773, 0.01888, 0.00944 and 0.00472 mg a.i./kg soil resulted in no significant effects of seedling emergence, survival, shoot length and plant biomass and therefore the NOEC was the highest rate tested of 0.1507 mg a.i./kg soil in this study and the LOEC > this rate. Symptoms of phytotoxicity were not apparent at any dose rate and there was no adverse effect on plant growth.

Ryegrass (Lolium perenne)

The foliar application of the test item applied at seven dose rates of 0.03773, 0.01888, 0.00944 0.00472, 0.00236, 0.001187, 0.0006 mg a.i./kg soil resulted in no significant effects of seedling emergence, survival, shoot length and plant biomass and therefore the NOEC was the highest rate tested in this study and the LOEC > this rate. LOEC and NOECs for both shoot length and biomass were statistically determined as 0.001187 and 0.0006 respectively however, the 0.001187 mg a.i./kg soil data is considered an outlier as rates from 0.00944 up to 0.03773 mg a.i./kg soil resulted in no significant adverse effect and therefore the values are considered >0.03773 and 0.03773 kg product/ha. Symptoms of phytotoxicity were apparent at the highest dose rate of 0.03773 mg a.i./kg soil and there was no adverse effect on plant growth.

Validity criteria fulfilled:

yes

Remarks:

See Table 1 in "Any other information on results incl. tables".

Reference 2

Endpoint:

toxicity to terrestrial plants: long-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

23 April - 18 May 2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

other: OECD (Draft Document): Guideline for the testing of chemicals, Proposal for updating Guideline 208 „Terrestrial (Non-Target) Plant Test: 208 A: Seedling Emergence and Seedling Growth Test. 208 B: Vegetative Vigour Test."

Version / remarks:

Part B, Draft Document July 2000

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPP 123-1 (Vegetative Vigor Tier II)

Version / remarks:

1982/Plants, PB83-153940; EPA540/9-82-020; Series 123

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Hessisches Ministerium für Umwelt, ländlichen Raum und Verbraucherschutz

Analytical monitoring:

yes

Details on sampling:

- Concentrations: A sample of the highest application rate was collected at day 0 and analysed for the test item.

Vehicle:

no

Details on preparation and application of test substrate:

- Method of application: Spray solution was applied once, at test initiation on the leaves and above-ground portions of plant; Plants were grown from seed to the 2 to 4-leaf stage. The test item was dissolved in deionized water and was applied once with 100 L/ha using an spray chamber equipped with an overhead nozzle (Lechner LU 367-02), with nozzle height set at 50 cm above the sprayed surface.
- Controls: The blank control spray solution was deionized water.

Species:

Lolium perenne

Plant group:

Monocotyledonae (monocots)

Details on test organisms:

- Common name: Perennial ryegrass
- Plant family: Poaceae
- Variety: Deutsches Weidegras
- Source of seed: from commercial sources via Bayer CropScience GmbH, Horticulture, H872, 65926 Frankfurt am Main; Germany
- Prior seed treatment/sterilization: Seeds used on the study had not been treated with pesticides or repellents prior to test initiation
- Seed storage: Seeds were stored in plastic boxes in the refrigerator

Species:

Allium cepa

Plant group:

Monocotyledonae (monocots)

Details on test organisms:

- Common name: Onion
- Plant family: Liliaceae
- Variety: Zittauer Gelbe
- Source of seed: from commercial sources via Bayer CropScience GmbH, Horticulture, H872, 65926 Frankfurt am Main; Germany
- Prior seed treatment/sterilization: Seeds used on the study had not been treated with pesticides or repellents prior to test initiation
- Seed storage: Seeds were stored in plastic boxes in the refrigerator

Species:

Avena sativa

Plant group:

Monocotyledonae (monocots)

Details on test organisms:

- Common name: Oat
- Plant family: Poaceae
- Variety: Flämings Nova
- Source of seed: from commercial sources via Bayer CropScience GmbH, Horticulture, H872, 65926 Frankfurt am Main; Germany
- Prior seed treatment/sterilization: Seeds used on the study had not been treated with pesticides or repellents prior to test initiation
- Seed storage: Seeds were stored in plastic boxes in the refrigerator

Species:

Zea mays

Plant group:

Monocotyledonae (monocots)

Details on test organisms:

- Common name: Corn
- Plant family: Poaceae
- Variety: Lorenzo
- Source of seed: from commercial sources via Bayer CropScience GmbH, Horticulture, H872, 65926 Frankfurt am Main; Germany
- Prior seed treatment/sterilization: Seeds used on the study had not been treated with pesticides or repellents prior to test initiation
- Seed storage: Seeds were stored in plastic boxes in the refrigerator

Species:

Lycopersicon esculentum

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

- Common name: Tomato
- Plant family: Solanaceae
- Variety: Balkonstar
- Source of seed: from commercial sources via Bayer CropScience GmbH, Horticulture, H872, 65926 Frankfurt am Main; Germany
- Prior seed treatment/sterilization: Seeds used on the study had not been treated with pesticides or repellents prior to test initiation
- Seed storage: Seeds were stored in plastic boxes in the refrigerator

Species:

Helianthus annuus

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

- Common name: Sunflower
- Plant family: Asteraceae
- Variety: Big Smile
- Source of seed: from commercial sources via Bayer CropScience GmbH, Horticulture, H872, 65926 Frankfurt am Main; Germany
- Prior seed treatment/sterilization: Seeds used on the study had not been treated with pesticides or repellents prior to test initiation
- Seed storage: Seeds were stored in plastic boxes in the refrigerator

Species:

Beta vulgaris

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

- Common name: Sugar beet
- Plant family: Chenopodiaceae
- Variety: Achat
- Source of seed: from commercial sources via Bayer CropScience GmbH, Horticulture, H872, 65926 Frankfurt am Main; Germany
- Prior seed treatment/sterilization: Seeds used on the study had not been treated with pesticides or repellents prior to test initiation
- Seed storage: Seeds were stored in plastic boxes in the refrigerator

Species:

Glycine max (G. soja)

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

- Common name: Soybean
- Plant family: Fabaceae
- Variety: Trail
- Source of seed: from commercial sources via Bayer CropScience GmbH, Horticulture, H872, 65926 Frankfurt am Main; Germany
- Prior seed treatment/sterilization: Seeds used on the study had not been treated with pesticides or repellents prior to test initiation
- Seed storage: Seeds were stored in plastic boxes in the refrigerator

Species:

Brassica napus

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

- Common name: Oilseed rape
- Plant family: Brassicaceae
- Variety: Liratop
- Source of seed: from commercial sources via Bayer CropScience GmbH, Horticulture, H872, 65926 Frankfurt am Main; Germany
- Prior seed treatment/sterilization: Seeds used on the study had not been treated with pesticides or repellents prior to test initiation
- Seed storage: Seeds were stored in plastic boxes in the refrigerator

Species:

Cucumis sativus

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

- Common name: Cucumber
- Plant family: Curcubitaceae
- Variety: Delikatess
- Source of seed: from commercial sources via Bayer CropScience GmbH, Horticulture, H872, 65926 Frankfurt am Main; Germany
- Prior seed treatment/sterilization: Seeds used on the study had not been treated with pesticides or repellents prior to test initiation
- Seed storage: Seeds were stored in plastic boxes in the refrigerator

Test type:

vegetative vigour test

Study type:

laboratory study

Substrate type:

natural soil

Limit test:

no

Total exposure duration:

21 d

Test temperature:

23 ± 5 °C at day and 15 ± 5°C night (greenhouse conditions)

pH:

7.4

Moisture:

Humidity: 80% day, 95% night (glasshouse control settings)

Details on test conditions:

TEST SYSTEM
- Testing facility: Bayer CropScience, Germany
- Test container: plastic container: 13 cm diameter x 10 cm high; 10 cm diameter and 9 cm high (onion only)
- Method of seeding: Seeds were placed into soil; Depth varied with size of seed, i.e. a 5 mm diameter seed was covered with 5 mm depth of soil, a 3 mm diameter see with 3 mm depth of soil
- No. of seeds per container: 40 seeds per treatment/10 species
- No. of plants (retained after thinning): 4 plants/pot: cucumber, sunflower, tomato 5 plants/pot: oilseed rape, soybean, sugar beet, corn, oat, onion, ryegrass
- No. of replicates per treatment group: 8 or 10 replicates
- No. of replicates per control: 8 or 10 replicates

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Geographic location: Standard soil (silty loam) from Bayer CropScience GmbH, Global Biology Herbicides, Horticulture, H 872, Industriepark Hoechst, 65926 Frankfurt am Main, Germany
Soil texture:
- % sand: 14.2%
- % silt: 65.1%
- % clay: 20.7%
- Soil taxonomic classification: Silty loam
- Organic carbon (%): 1.19%
- pH: 7.4
- Pretreatment of soil: Locally supplied soil steam pasteurized before use (120 °C vapour for about 30 minutes)

NUTRIENT MEDIUM
- Description: Two applications of 0.1% liquid fertilizer (Wurax Super) with sub-irrigation during the test period.

GROWTH CONDITIONS
- Photoperiod: 16 h light, 8 h dark (glasshouse control settings)
- Light intensity and quality: Natural daylight supplemented by artificial lighting to provide the required photoperiod. Regulation of light intensity > 10000 lux lamps turn off, > 20000 lux shading closing
- Day/night temperatures: 23 ± 5 °C day, 15 ± 5 °C night (temperature control set)
- Relative humidity (%): 80% day, 95% night (glasshouse control settings)
- Watering regime and schedules: Following the initial top watering to facilitate germination of the seed, bottom watering was performed with saucers standing below each pot. Water was given and retained within the saucer according to the need of the plants in order to have an optimal water supply for plant growth. This was checked daily.
- Water source/type: Local tap water. Watered as required at least once per day. Pots contained in plastic saucers in which the water was added
- Interval of applications: once at test start
- Method of application: applied once with 100 L/ha using an spray chamber equipped with an overhead nozzle (Lechner LU 367-02), with nozzle height set at 50 cm above the sprayed surface
- Any pest control method/fertilization: 2.4 g/L of granular fertilizer (Blaukorn) was added to the soil prior to sowing

EFFECT PARAMETERS MEASURED: Survival, phytotoxicity, growth stage, plant dry weight and length; visual enumeration; total shoot length to 0.1 cm; dry weight per balance to 0.001 g; Observation interval: weekly.

- Phytotoxicity rating system: 0 (no injury/effect), 20% (indicates slight effect), 40% (indicates moderate effect), 60% (indicates severe effect), 80% (Indicates total plant effect), 100% (plant death)

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2

Nominal and measured concentrations:

Control, 0.227, 0.453, 0.906, 1.77, 3.53, 7.07, 14.2, 28.3, 56.6 and 113 g test item/ha (nominal application rate)
Control, 0.005, 0.01, 0.02, 0.039, 0.078, 0.156, 0.313, 0.625, 1.25, and 2.5 kg product/ha (nominal test concentrations)
Control, 2.5, 1.25, 0.625, 0.313 and 0.156 kg product / ha (Perennial ryegrass and oats)
Control, 2.5, 1.25, 0.625, 0.313, 0.156 and 0.078 kg product / ha (onion and corn)
Control, 0.156, 0.078, 0.039, 0.020, 0.010 and 0.005 kg product / ha (Cucumber, oilseed rape, soybean, sunflower, sugar beet and tomato)

Reference substance (positive control):

no

Species:

Beta vulgaris

Duration:

21 d

Dose descriptor:

EC50

Effect conc.:

0.003 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: plant biomass (shoot dry weight)

Remarks on result:

other: equivalent to 2.36 g a.i./ha

Species:

Beta vulgaris

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.001 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: plant biomass (shoot dry weight)

Remarks on result:

other: equivalent to 0.45 g a.i./ha

Species:

other: Brassica napus, Beta vulgaris, Cucumis sativus

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.001 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: Plant shoot length

Remarks on result:

other: eqivalent to 0.45 g a.i./ha

Species:

Lycopersicon esculentum

Duration:

21 d

Dose descriptor:

EC50

Effect conc.:

0.004 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: Plant Shoot length

Remarks on result:

other: equivalent to 2.81 g a.i./ha

Species:

Beta vulgaris

Duration:

21 d

Dose descriptor:

EC50

Effect conc.:

0.006 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: Plant survival

Remarks on result:

other: equivalent to 4.8 g a.i./ha

Species:

Lycopersicon esculentum

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.002 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: Plant survival

Remarks on result:

other: equivalent to 1.77 g a.i./ha

Details on results:

- Typical symptoms: bleaching, chlorosis, necrosis and growth suppression and all species except for ryegrass exhibited some or all of these symptoms, the severity of which increased with dose rate tested.
- Biological and analytical results in details are summarized with the tables in section "Any other information on results incl tables"

Reported statistics and error estimates:

Mortality, shoot length and biomass (plant dry weight) were compared using the ToxRat software for statistical analysis (version 2.09).

Endpoints:

The endpoints were reported as kg test item/ha and g a.i./ha. Based on these reported values the endpoints were recalculated to mg/kg soil which were not part of the report but based on the reported data of g a.i./h and a density of 1.5 g/cm³ and a soil depth of 5 cm.

Analytical Results:

Measured concentrations were 95.7% of nominal.

Table 1: The mean phytotoxicity values at day 21 for the vegetative vigour test with the test item

Day 21 phytotoxicity summary (mean % damage*) Dicot species
mg a.i./kg soil	Cucumber	Oilseed rape	Soybean	Sugar beet	Sunflower	Tomato
0.00944	41	65	55	86	55	95
0.00472	15	40	15	28	12	95
0.00236	15	15	15	15	0	35
0.001187	0	0	0	0	0	0
0.0006	0	0	0	0	0	0
0.0003067	0	0	0	0	0	0
Control	0	0	0	0	0	0

 

Day 21 phytotoxicity summary (mean % damage) Monocot species

mg a.i./kg soil	Corn	Oat	Onion	Ryegrass
0.1507	70	15	75	0
0.07551	35	0	15	0
0.03773	0	0	0	0
0.01888	0	0	0	0
0.0.00944	0	0	0	0
0.00472	0	NT	0	NT
Control	0	0	0	0

* % effect values were calculated in comparison to controls.

NT = concentration not tested

Table 2: Analytical Results

Sample no.	Desired value mg/L	Result mg/L	Result % of theory	CV (%)
Recovery	1091.20	1100.59	100.9	0.2
VV04/005-1-001 (Control)	0	0	--	--
VV04/005 -1-002 (Stock solution)	1132.50	1083.96	95.7	0.8

Table 3: Validity criteria for OECD 208

Criterionfrom the guideline	Outcome	Validity criterion fulfilled
Seedling emergence in the controls is ≥ 70%.	-	-
No visible phytotoxic effects (e.g. chlorosis, necrosis, wilting, deformatons) in control seedlings and the plants of every particular species exhibit normal variation in growth and morphology.	No visible phytotoxic effects occurred in the controls.	yes
In the controls, the mean survival of emerged control seedlings is ≥ 90% for the duration of the study.	In the controls, the mean survival of emerged control seedlings was ≥ 97.5% for the duration of the study.	yes
The environmental conditions for a particular species are identical and the growing media contain the same amount of soil matrix, support media or substrate from the same source.	The environmental conditions for a particular species were identical and the growing media contain the same amount of soil matrix, support media or substrate from the same source.	yes

 

Dicotyledonous species

Cucumber (Cucumis sativus)

The foliar application of the test item applied at six dose rates of 0.0003067, 0.0006, 0.001187, 0.00236, 0.00472 and 0.00944 mg a.i./kg soil resulted in no significant effects of survival and therefore the NOEC was the highest rate tested in this study and the LOEC > than this rate. Analysis of shoot length after 21 days resulted in EC25 and EC50 endpoints of 0.00163 and 0.00944 mg a.i./kg soil, respectively. Significant adverse effects on shoot length were apparent at dose rates including and above 0.001187 mg a.i./kg soil with a NOEC, determined by Dunnett’s t-test of 0.0006 mg a.i./kg soil. Analysis of plant biomass (shoot dry weight) at day 21 resulted in EC25 and EC50 endpoints both >0.00944 mg a.i./kg soil, with significant adverse effects at 0.00944 mg a.i./kg soil and a NOEC, determined by Dunnett’s t-test of 0.00472 mg a.i./kg soil. Symptoms of phytotoxicity were apparent at dose rates including and above 0.00236 mg a.i./kg soil and there was no impact on the development of plant growth as determined by growth stage analysis at day 21.

 

Oilseed rape (Brassica napus)

The foliar application of the test item applied at six dose rates of 0.0003067, 0.0006, 0.001187, 0.00236, 0.00472 and 0.00944 mg a.i./kg soil resulted in no significant effects of survival and therefore the NOEC was the highest rate tested in this study and the LOEC > this rate. Analysis of shoot length after 21 days resulted in EC25 and EC50 endpoints of 0.00373 and 0.00857 mg a.i./kg soil, respectively. Significant adverse effects on shoot length were apparent at dose rates including and above 0.001187 mg a.i./kg soil with a NOEC, determined by William’s t-test of 0.0006 mg a.i./kg soil. Analysis of plant biomass (shoot dry weight) at day 21 resulted in EC25 and EC50 endpoints of 0.000229 and 0.00405 mg a.i./kg soil, respectively. Significant adverse effects on shoot biomass were apparent at dose rates including and above 0.00236 mg a.i. /kg soil with a NOEC, determined by William’s t-test, of 0.001187 mg a i/kg soil. Symptoms of phytotoxicity were apparent at dose rates including and above 0.00236 mg a.i./kg soil. Plant growth development as determined by growth stage analysis at day 21 was reduced at dose rates including and above 0.001187 mg a.i. /kg soil.

 

Soybean (Glycine max)

The foliar application of the test item applied at six dose rates of 0.0003067, 0.0006, 0.001187, 0.00236, 0.00472 and 0.00944 mg a.i./kg soil resulted in no significant effects of survival and therefore the NOEC was the highest rate tested in this study and the LOEC > this rate. Analysis of shoot length after 21 days resulted in EC25 and EC50 endpoints of 0.00852 and > 0.00944 mg product/ha, respectively. Significant adverse effects on shoot length were apparent at dose rates including and above 0.00236 mg a.i./kg soil with a NOEC, determined by William’s t-test of 0.001187 mg product/ha. Analysis of plant biomass (shoot dry weight) at day 21 resulted in EC25 and EC50 endpoints of 0.0026 and 0.005253 mg product/ha, respectively. Significant adverse effects on shoot biomass were apparent at dose rates including and above 0.00236 mg product/ha with a NOEC, determined by William’s t-test, of 0.001187 mg product/ha. Symptoms of phytotoxicity were apparent at dose rates including and above 0.00236 mg product/ha and there was no impact on the development of plant growth as determined by growth stage analysis at day 21.

 

Sugar beet (Beta vulgaris)

The foliar application of the test item applied at six dose rates of 0.0003067, 0.0006, 0.001187, 0.00236, 0.00472 and 0.00944 mg a.i./kg soil resulted in EC25 and EC50 endpoints for survival of 0.082 and 0.106 kg product/ha, repectively. Analysis of shoot length after 21 days resulted in EC25 and EC50 endpoints of 0.0026 and 0.00532 mg a.i./kg soil, respectively. Significant adverse effects on shoot length were apparent at dose rates including and above 0,001186667 kg product/ha with a NOEC, determined by William’s t-test of 0.0006 mg a.i./kg soil. Analysis of plant biomass (shoot dry weight) at day 21 resulted in EC25 and EC50 endpoints of 0,001866667 and 0,003146667 mg a.i./kg soil, respectively. Significant adverse effects on shoot biomass were apparent at dose rates including and above 0.001186667 mg a.i./kg soil with a NOEC, determined by William’s t-test, of 0,0006 mg a.i./kg soil. Symptoms of phytotoxicity were apparent at dose rates including and above 0.00236 mg a.i./kg soil. Development of plant growth as determined by growth stage analysis at day 21 was reduced at dose rates including and above 0.00236 mg a.i./kg soil.

 

Sunflower (Helianthus annuus)

The foliar application of the test item applied at six dose rates of 0.0003067, 0.0006, 0.001187, 0.00236, 0.00472 and 0.00944 mg a.i./kg soil resulted in no significant effects of survival and therefore the NOEC a was the highest rate tested in this study and the LOEC > this rate. Analysis of shoot length after 21 days resulted in EC25 and EC50 endpoints of 0.005373 and 0.008213 mg a.i./kg soil, respectively. Significant adverse effects on shoot length were apparent at dose rates including and above 0.00236 mg a.i./kg soil with a NOEC, determined by William’s t-test of 0.001187 mg a.i./kg soil. Analysis of plant biomass (shoot dry weight) at day 21 resulted in EC25 and EC50 endpoints of 0.004413 and 0.00616 mg a.i./kg soil, respectively. Significant adverse effects on shoot biomass were apparent at dose rates including and above 0.00472 mg a.i./kg soil with a NOEC, determined by William’s t-test, of 0.00236 mg a.i./kg soil. Symptoms of phytotoxicity were apparent at dose rates including and above 0.00472 mg a.i./kg soil. Development of plant growth as determined by growth stage analysis at day 21 was reduced at the highest dose rate tested of 0.00944 mg a.i./kg soil.

Tomato (Lycopersicum esculentum)

The foliar application of the test item applied at six dose rates of 0.0003067, 0.0006, 0.001187, 0.00236, 0.00472 and 0.00944 mg a.i./kg soil resulted in EC25 and EC50 endpoints of 0.086 and 0.137 kg product/ha, respectively with a NOEC, determined by the Fishers exact Binomial test with Bonferroni correction of 0.00236 mg a.i./kg soil. Analysis of shoot length after 21 days resulted in EC25 and EC50 endpoints of 0.00212 and 0.003747 mg a.i./kg soil, respectively. Significant adverse effects on shoot length were apparent at dose rates including and above 0,00236 mg a.i./kg soil with a NOEC, determined by William’s t-test of 0.001187 mg a.i./kg soil. Analysis of plant biomass (shoot dry weight) at day 21 resulted in EC25 and EC50 endpoints of 0.001627 and 0.00327 mg a.i./kg soil, respectively. Significant adverse effects on shoot biomass were apparent at dose rates including and above 0.00236 mg a.i./kg soil with a NOEC, determined by William’s t -test, of 0.001187 mg a.i./kg soil. Symptoms of phytotoxicity were apparent at dose rates including and above 0.00236 mg a.i./kg soil. Plant growth as determined by growth stage analysis at day 21 was reduced at dose rates including and above 0.001187 mg a.i./kg soil.

Monocotyledonous species

Corn (Zea mays)

The foliar application of the test item applied at six dose rates of 0.00472, 0.00944, 0.01888, 0.03773, 0,07551 and 0,1507 mg a.i./kg soil resulted in no significant effects of survival and therefore the NOEC the highest rate tested in this study and the LOEC > this rate. Analysis of shoot length after 21 days resulted in EC25 and EC50 endpoints both >0.1507 mg a.i./kg soil. Significant adverse effects on shoot length were apparent at dose rates including and above 0.07551 mg a.i./kg soil with a NOEC of 0.03773 mg a.i./kg soil. Analysis of plant biomass (shoot dry weight) at day 21 resulted in EC25 and EC50 endpoints of 0.06 and 0.1112 mg a.i./kg soil, respectively. Significant adverse effects on shoot biomass were apparent at dose rates including and above 0.07551 mg a.i./kg soil with a NOEC, determined by the Welch t-test, of 0.03773 mg a.i./kg soil. Symptoms of phytotoxicity were apparent at dose rates including and above 0.07551 mg a.i./kg soil. There was no impact in growth stages for all application rates.

 

Oats (Avena sativa)

The foliar application of the test item applied at five dose rates of 0.00944, 0.01888, 0.03773, 0.07551 and 0.1507 mg a.i./kg soil resulted in no significant effects of survival and therefore the NOEC was the highest rate tested in this study and the LOEC > this rate. Analysis of shoot length after 21 days resulted in EC25 and EC50 endpoints both >0.1507 mg a.i./kg soil. Significant adverse effects on shoot length were apparent at the highest dose rate of 0.1507 mg a.i./kg soil with a NOEC of 0.07551 mg a.i./kg soil. Analysis of plant biomass (shoot dry weight) at day 21 resulted in EC25 and EC50 endpoints both 0.07667 and >0.1507 mg a.i./kg soil, respectively. Significant adverse effects on shoot biomass were apparent at dose rates including and above 0.07551 mg a.i./kg soil with a NOEC, determined by William’s t-test, of 0.037733333 mg a.i./kg soil. Symptoms of phytotoxicity were apparent at the highest rate tested of 0.1507 mg a.i./kg soil and there was no adverse effect on the development of plant growth.

 

Onion (Allium cepa)

The foliar application of the test item applied at six dose rates of 0.00472, 0.00944, 0.01888, 0.03773, 0,07551 and 0,1507 mg a.i./kg soil resulted in EC25 and EC50 endpoints of 1.272 and 1.612 kg product/ha, respectively with a NOEC, determined by the Fishers exact Binomial test with Bonferroni correction of 0.075507 mg a.i. /kg soil. The LOEC was 0.1507 mg a.i. /kg soil. Analysis of shoot length after 21 days resulted in EC25 and EC50 endpoints of 0.0744 and >0.1507 mg a.i. /kg soil, respectively. Significant adverse effects on shoot length were apparent at dose rates including and above 0.07551 mg a.i./kg soil with a NOEC, determined by William’s ttest, of 0.03773 mg a.i. /kg soil. Analysis of plant biomass (shoot dry weight) at day 21 resulted in EC25 and EC50 endpoints both 0.0385 and 0.0947 mg a.i./kg soil, respectively. Significant adverse effects on shoot biomass were apparent at dose rates including and above 0.03773 mg a.i./kg soil with a NOEC, determined by William’s t-test, of 0.01888 mg a.i./kg soil. Symptoms of phytotoxicity were apparent at dose rates including and above 0.07551 mg a.i./kg soil and development of plant growth as determined by growth stage analysis at day 21 was reduced at the highest does rate of 0.1507 mg a.i. /kg soil.

 

Ryegrass (Lolium perenne)

The foliar application of the test item applied at five dose rates of 0.156, 0.313, 0.625, 1.25 and 2.5 mg a.i./kg soil resulted in no significant effects of survival and therefore the NOEC was the highest rate tested in this study and the LOEC was > this rate. Analysis of shoot length resulted EC25 and EC50 endpoints both >0.1507 mg a.i./kg soil. A NOEC was determined at 0.01888, however, there was a 16.4% increase in shoot length at this dose rate and only a 9.9% decrease in shoot length at 0.1507 mg a.i. /kg soil. There was no adverse effect of shoot biomass (shoot dry weight) up to and including the highest rate tested of 0.1507 mg a.i. /kg soil. There were also no symptoms of phytotoxicity or adverse effects on the development of plant growth at any dose rate.

Validity criteria fulfilled:

yes

Remarks:

See Table 3 in "Any other information on results incl. tables".

Description of key information

NOEC (21 d): 0.0006 mg a.i./kg soil (vegetative vigour, Sugar beet, Cucumber, Oilseed rape, OECD 208, shoot length,and biomass for sugar beet, nominal)

NOEC (21 d): 0.001187 mg a.i./kg soil (seedling emergence, Tomato, OECD 208, shoot length, nominal)

Key value for chemical safety assessment

Long-term EC10, LC10 or NOEC for terrestrial plants:

0.001 mg/kg soil dw

Additional information

Two experimental studies are available investigating the potential effects of the test item (CAS 874195-61-6) on vegetative vigour (M-263656-01-2) and Seedling emergence and growth (M-263640-01-2) of plants. The studies were performed according to OECD guideline 208 under GLP conditions. The substance tested was a mixture of the active substance and a safener. However, the effect concentrations of both studies were reported based on the active substance.

For the vegetative vigour study (M-263656-01-2) ten species of non-target terrestrial plants (4 monocots and 6 dicots) were sprayed at various application rates of the formulation. Perennial ryegrass and oats were sprayed with 0.00944, 0.01888, 0.03773, 0.07551 and 0.1507 mg a.i./kg soil. Onion and corn were sprayed with 0.00472, 0.00944, 0.01888, 0.03773, 0.07551 and 0.1507 mg a.i./kg soil. Cucumber, oilseed rape, soybean, sunflower, sugar beet and tomato were sprayed with 0.0003067, 0.0006, 0.001187, 0.00236, 0.00472 and 0.00944 mg a.i./kg soil. All plants were at the two to four leaf stages on the day of application and test duration was 21 days following application of the test substance. Spray treatments were applied once to each species at test initiation with a laboratory track sprayer set at the nominal spray volume of 100 litres/ha. Spray stock solutions were analysed for the quantification of test item. Measured concentrations in the spray solutions were 95.7% of nominal. A total of forty plants were used for each application rate. There were ten replicate pots of cucumber sunflower and tomato, with each pot containing four plants. There were eight replicates of the other seven species with each pot containing five plants. All pots were randomised on benches within the glasshouse. Typical symptoms with the test item observed in this study were bleaching, chlorosis, necrosis and growth suppression and all species except for ryegrass exhibited some or all of these symptoms, the severity of which increased with dose rate tested. The most sensitive EC50 was 0.003147 mg a.i./kg soil equivalent to 2.36 g a.i.(test item)/ha for sugar beet plant biomass. The lowest NOEC value was 0.0006 mg a.i./kg soil, equivalent to 0.45 g a.i./ha for shoot length (cucumber, oilseed rape and sugar beet) for shoot biomass (sugar beet).

For the seedling emergence and growth study (M-263640-01-2) ten species of non-target terrestrial plants (4 monocots and 6 dicots) were sprayed at various application rates of the formulation. Corn and oat were sprayed with 0.1507, 0.07551, 0.03773, 0.01888, 0.00944, 0.00472 mg a.i./kg soil. Barley was sprayed with 0.07551, 0.03773, 0.01888, 0.00944, 0.00472 and 0.00236 mg a.i./kg soil. Cucumber, oilseed rape, soybean, sunflower, sugar beet, tomato and ryegrass were sprayed with 0.037733333, 0.01888, 0.00944 0.00472, 0.00236, 0.001187, 0.0006 mg a.i./kg soil. Seeds were sown in soil in pots prior to application of the test substance. Spray treatments were applied once to the soil surface with an Track sprayer set at the nominal spray volume of 100 L/ha. Control pots were sprayed with deionised water. Spray stock solutions were analysed for the quantification of the test item. Measured concentrations of the exposure solution were 92.5% - 96.5% of nominal. A total of forty seeds were used for each treatment level. There were 8 replicates for all species with each pot containing 5 seeds. All pots were randomised on benches within the glasshouse. Visually observations for germination and phytotoxicity were made on test days 7, 14 and 21. Final assessments were made for germination (emergence) growth stage, plant survival, shoot height and shoot biomass (dry weight) 21 days after application against the untreated controls. Statistical analysis of data was performed to obtain NOEC, LOEC and EC50 values for emergence, survival shoot length and biomass (shoot dry weight), using ToxRat software for statistical analysis (version 2.09). This study was valid with NOECs obtained for all 10 species and with all endpoints.. The most sensitive EC50 was 0.00816 mg a.i./kg soil equivalent to 6.12 g a.i.(test item)/ha for tomato plant biomass. The lowest NOEC was 0.001187 mg a.i./kg soil, equivalent to 0.89 g a.i./ha for tomato shoot length.