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Toxicity to terrestrial plants

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Reference
Endpoint:
toxicity to terrestrial plants: long-term
Type of information:
experimental study
Adequacy of study:
key study
Study period:
26 Apr - 25 May 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 208 (Terrestrial Plants, Growth Test)
Deviations:
yes
Remarks:
Temperature and humidity were outside of the recommended range owing to a technical fault. They are not considered to have had any relevant impact on the test results. This judgement is supported by the condition and vitality of the control group.
GLP compliance:
yes (incl. QA statement)
Remarks:
Sächsisches Staatsministerium für Umwelt und Landwirtschaft, Dresden, Germany
Analytical monitoring:
no
Vehicle:
no
Details on preparation and application of test substrate:
- Method of mixing into soil (if used): After cooling the test item in a refrigerator for 2 hours at <= -18 ºC, the test item was grated to a fine powder using a grater. It was ensured that not too much force was applied during the grating to prepare a very fine powder. The grating procedure was performed in a cooled room at 2-8 ºC.
Since the test item is insoluble in water, an exact weighed test item amount was mixed with a defined amount of soil. This mixture was prepared immediately prior to application. The mixture was diluted with soil so that 1000 g of the dilution contained the amount of test item required for the test item concentration of 1000 mg test item/kg soil dry weight. The treated soil (1000 g) was added to the prepared amount of soil (24 kg dry weight) yielding 25 kg dry soil. The test item mixture was then mixed thoroughly with the soil by intensive stirring in a mixing machine at 2-8 ºC. Before application, the soil was cooled down to 2-8 ºC. The cooled soil inhibited melting and agglutinating of the test item powder within the soil during application. The homogeneity was assessed visually.
- Controls: yes, test soil without application of test item
Species:
Avena sativa
Plant group:
Monocotyledonae (monocots)
Details on test organisms:
- Plant family: Poaceae
- Variety: Scorpion
- Source of seed: Saatenunion GmbH, Isernhagen, Germany
- Historical germination of seed (germination of seed lot tested): 91%
Species:
Allium cepa
Plant group:
Monocotyledonae (monocots)
Details on test organisms:
- Plant family: Liliaceae
- Variety: Braunschweiger Dkl. blutrote
- Source of seed: N.L. Chrestensen, Erfurt, Germany
- Historical germination of seed (germination of seed lot tested): 94%
Species:
Brassica napus
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Plant family: Brassicaceae
- Variety: Visby
- Source of seed: Saatenunion GmbH, Isernhagen, Germany
- Historical germination of seed (germination of seed lot tested): 95%
Species:
Lycopersicon esculentum
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Plant family: Solanaceae
- Variety: Tamina
- Source of seed: N.L. Chrestensen, Erfurt, Germany
- Historical germination of seed (germination of seed lot tested): 89%
Species:
Cucumis sativus
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Plant family: Cucurbitaceae
- Variety: Burpless Tasty Green
- Source of seed: N.L. Chrestensen, Erfurt, Germany
- Historical germination of seed (germination of seed lot tested): 96%
Species:
Glycine max (G. soja)
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Plant family: Fabaceae
- Variety: Merlin
- Source of seed: Saatbau Linz, Leonding, Austria
- Historical germination of seed (germination of seed lot tested): 98%
Test type:
seedling emergence and seedling growth test
Study type:
laboratory study
Substrate type:
natural soil
Limit test:
yes
Total exposure duration:
21 d
Remarks:
21 d after 50 % of the control plants had emerged.
Test temperature:
15.3 - 35.6 °C
pH:
6.3
Details on test conditions:
TEST SYSTEM
- Testing facility: BioChem agrar, Gerichshain, Germany
- Test container (type, material, size): non-porous plastic flower pot (diameter: 15 cm)
- Amount of soil: 1 kg dry soil/pot
- Method of seeding: The seeds were sown on the soil surface and covered with a thin layer of soil.
- No. of seeds per container: 5 (oat, onion), 3 (oilseed rape), 2 (tomato, cucumber, soybean); number of seeds/treatment: 20-25
- No. of replicates per treatment group: 5 (oat, onion), 7 (oilseed rape), 10 (tomato, cucumber, soybean)
- No. of replicates per control: 5 (oat, onion), 7 (oilseed rape), 10 (tomato, cucumber, soybean)

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Geographic location: agricultural soil from site Gerichshain (batch G 01/2016)
- Pesticide use history at the collection site: no pesticides were applied on the origin plot of the test soil for at least 5 years (the plot was fallow land)
- Soil texture (if natural soil)
- % sand: 58.3%
- % silt: 30.7%
- % clay: 11.0%
- Soil taxonomic classification: loamy sand
- Soil classification system: DIN 4220
- Organic carbon (%): 0.9%
- Maximum water holding capacity (in % dry weigth): 28.0 g/100 g dry soil
- Salt content (mg KCl/100 g dry soil): 37.2
- pH: 6.3
- Pretreatment of soil: sieved before use in the test
- Storage (condition, duration): stored for at least 1 year

GROWTH CONDITIONS
- Photoperiod: 16 h light (incl. 4 h HQL light)
- Light intensity and quality: 311 - 391 μE/m²/s
- Day/night temperatures: 15.3 - 35.6 °C
- Relative humidity (%): 20.2 - 86.4%
- Watering regime and schedules: bottom watering in pot saucers daily with tap water (pH: 7.3, conductivity: 80 µS/cm)
- Volume applied: The amount of water depended on the growth of the plants.
- Any pest control method/fertilization (if used): according to good horticultural practice with the liquid fertiliser “Pflanzenvollnahrung“ (origin: Gute Wahl GmbH, München, Germany); content: 4% N, 3% P2O5, 3% K2O; cycle: weekly; amount/pot: 100 mL (0.5%)

EFFECT PARAMETERS MEASURED
- 7 to 21 days after 50% seedling emergence in the control group (weekly), the plants were observed for emergence (mortality), survival of emerged seedlings and visible phytotoxicity compared to untreated control plants
- Determination of shoot fresh weight after 21 d
- Determination of growth stage after 7, 14 and 21 d

General assessment parameters
- mortality (photosynthetically inactive plants, wilted plants)
- plant growth (shoot fresh weight, growth inhibition)
- chlorosis (full leaf, partially on the leaf, between lead veins, albinism)
- necrosis (marginal, apical, isolated on the leaf, between leaf veins, on root crown)
- deformation (leaf: ruffled, coiled, not unfolded, dwarf stem)
- alteration in habitus (parted shoot apex, swollen leaf veins, dwarf, leaf veins deformed, compressed internodes, wilting)

VEHICLE CONTROL PERFORMED: no (not applicable)

TEST CONCENTRATIONS
- Spacing factor for test concentrations: not applicable (limit test)
Nominal and measured concentrations:
nominal: control, 1000 mg/kg soil dw
Reference substance (positive control):
no
Species:
other: Avena sativa, Allium cepa, Brassica napus, Lycopersicon esculentum, Cucumis sativus, Glycine max
Duration:
21 d
Dose descriptor:
EC10
Remarks:
EC50
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
seedling emergence
Species:
other: Avena sativa, Allium cepa, Brassica napus, Lycopersicon esculentum, Cucumis sativus, Glycine max
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
>= 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
seedling emergence
Species:
other: Avena sativa, Allium cepa, Brassica napus, Lycopersicon esculentum, Cucumis sativus, Glycine max
Duration:
21 d
Dose descriptor:
EC10
Remarks:
EC50
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: survival of emerged seedlings
Species:
other: Avena sativa, Allium cepa, Brassica napus, Lycopersicon esculentum, Cucumis sativus, Glycine max
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
>= 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: survival of emerged seedlings
Species:
other: Avena sativa, Allium cepa, Brassica napus, Lycopersicon esculentum, Cucumis sativus, Glycine max
Duration:
21 d
Dose descriptor:
EC10
Remarks:
EC50
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: shoot fresh weight
Species:
other: Avena sativa, Allium cepa, Brassica napus, Lycopersicon esculentum, Cucumis sativus, Glycine max
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
>= 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: shoot fresh weight
Details on results:
SEEDLING EMERGENCE
- Percent seedling emergence: 100% after 21 d in the control and the treatment for all plant species
- Percent survival: 100% after 21 d in the control and the treatment for all plant species
- Chlorosis: 0%
- Necrosis: 0%
- BBCH growth stage: No changes between control and treatment were recorded for all plant species.
- Shoot fresh weight: For all plant species the inhibition of shoot fresh weigth was < 5% compared to the control.

SEED GERMINATION
- Percent seed germination: 98-98%
Reported statistics and error estimates:
Effects on seedling emergence (inhibition in %) and survival (mortality in %), effects on shoot fresh weight (mean, SD, cv and % reduction) and phytotoxic effects (necrosis, chlorosis and growth inhibition in %) were calculated for each plant species and treatment group. Statistical analysis was performed using the software ToxRat Professional 3.2.1 (Ratte, 2015).
For statistical evaluation of the seedling emergence/survival of emerged seedlings results Fisher`s Exact Binomial Test (alpha = 0.05, one-sided greater) was used. For the biomass (shoot fresh weight) the NOEC values were determined using the Student-t-test (alpha = 0.05, two-sided).
Validity criteria fulfilled:
yes

Description of key information

Experimental result: Overall NOEC (21 d) ≥ 1000 mg/kg soil dw (OECD 208)

Key value for chemical safety assessment

Additional information

One experimental study is available investigating the effects of UMA 121 on the seedling emergence and growth of six plant species (Friedrich, 2016). The study was conducted according to OECD 208 (GLP) using two monocotyledonous (Avena sativa and Allium cepa) and four dicotyledonous (Brassica napus, Lycopersicon esculentum, Cucumis sativus and Glycine max) plant species. UMA 121 was ground to a very fine powder and directly incorporated into soil. A natural agricultural soil (loamy sand) with 0.9% organic carbon was used for testing. The study was designed as a limit test with a nominal concentration of 1000 mg/kg soil dw and 20-25 seeds/plant species. The temperature and humidity were outside of the recommended range owing to a technical fault. They are not considered to have had any relevant impact on the test results. This judgement is supported by the condition and vitality of the control group. During the study the seedling emergence, survival of emerged seedlings and phytotoxicity compared to the untreated control was assessed. Moreover the shoot fresh weight and growth stage were recorded. The study was finalised 21 d after 50% of the control plants had emerged. No effects on any plant species were observed resulting in an EC10/EC50 (21 d) > 1000 mg/kg soil dw and a NOEC (21 d) ≥ 1000 mg/kg soil dw.