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Toxicity to terrestrial plants

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Reference
Endpoint:
toxicity to terrestrial plants: short-term (with study design considered suitable for long-term assessment)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2014-04-08 to 2014-04-29
Reliability:
1 (reliable without restriction)
Qualifier:
according to
Guideline:
OECD Guideline 208 (Terrestrial Plants Test: Seedling Emergence and Seedling Growth Test)
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
no
Details on sampling:
No analytical monitoring
Vehicle:
no
Details on preparation and application of test substrate:
- Method of mixing into soil (if used): The respective test item amount was weighed out for each test item concentration and mixed thoroughly with
quartz sand in a glass vessel (1 % of the entire soil amount). Afterwards the treated quartz sand was mixed carefully with the soil using a mixer to
ensure a homogeneous distribution of the test item. The treated soil was transferred to the test container and the seeds were sown.
- Control: Untreated soil with 1 % quartz sand
Species:
Brassica napus
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Common name: Rape
- Plant family: Brassicaceae
- Variety: Kadore
- Source of seed: KWS Mais GmbH, D-37555 Einbeck, Germany
- Prior seed treatment/sterilization: No
- Seed storage: The seeds were stored dark at the test facility at room temperature (22 ± 10 °C) and protected from moisture until use.
Species:
Avena sativa
Plant group:
Monocotyledonae (monocots)
Details on test organisms:
- Common name: Oats
- Plant family: Gramineae
- Variety: Ivory
- Source of seed: NORDSAAT SAATZUCHT GMBH, 38895 Langenstein, Germany
- Prior seed treatment/sterilization: No
- Seed storage: The seeds were stored dark at the test facility at room temperature (22 ± 10 °C) and protected from moisture until use.
Species:
Glycine max (G. soja)
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Common name: Soybean
- Plant family: Leguminosae
- Variety: Merlin
- Source of seed: Saatbau Linz, A-4021 Linz, Austria
- Prior seed treatment/sterilization: No
- Seed storage: The seeds were stored dark at the test facility at room temperature (22 ± 10 °C) and protected from moisture until use.
Test type:
seedling emergence toxicity test
Study type:
laboratory study
Substrate type:
natural soil
Limit test:
no
Total exposure duration:
21 d
Test temperature:
nominal: 22 +/- 10 °C
actual: 13 - 21 °C
pH:
5.31 +/- 0.01 (natural soil)
Moisture:
- relative humidity: nominal: 70 +/- 25 %, actual: 58 - 100 %
Details on test conditions:
TEST SYSTEM
- Testing facility: Dr.U.Noack-Laboratorien
- Test container (type, material, size): Plastic container (standard flower pots) with a diameter of ca. 12 cm
- Amount of soil:
Control and 1000 mg/kg DW: 16.94 kg, divided into two parts, each 8.47 kg
400 - 160 - 64 - 25.6 mg/kg DW: 12.72 kg, divided into two parts, each 6.36 kg
- Method of seeding: Test containers were filled each with the same amount of treated soil (see application technique) and seeding holes were made
with a seedling pistil. One seed was given in each hole and covered with soil. During cultivation the test containers were bottom watered and
fertilized as necessary.
- No. of seeds per container: 5
- No. of replicates per treatment group: 8
- No. of replicates per control: 8


SOURCE AND PROPERTIES OF SUBSTRATE (if soil): A 2:1 mixture of natural soil LUFA 2.2 (batch number: Sp2.20214) and quartz sand (12a) was used.
- Geographic location: Soil collected by LUFA Speyer (67346 Speyer, Germany), quartz sand collected by Dörentrup Quarz GmbH Co. KG (31089
Duingen, Germany)
- Pesticide use history at the collection site: No pesticide use for the last five years.
- Soil texture (if natural soil)
- grain size: - Soil taxonomic classification: loamy sand (IS)
- Soil classification system: German DIN
- Organic carbon (%): 0.73
- CEC: 9.9 +/- 0.7 mval/100 g
- Storage (condition, duration): The soil has been stored at the test facility at green house conditions until use.


NUTRIENT MEDIUM (if used)
- Description:
Fertilizer: HAKAPHOS® SOFT SPEZIAL 16+8+22(+3) (0.5 ‰)
Origin: GRÜNES LANDHAUS, St. Godehard-Str. 23, 31139 Hildesheim, Germany

Nutrient Concentration [%]
Ammonium nitrogen 9.7
Nitrate nitrogen 6.3
P2O5 8
Potassium oxide 22
Magnesium oxide 3
Boron 0.03
Iron (Chelate) 0.15
Copper 0.02
Manganese 0.05
Molybdenum 0.005
Zinc 0.015


GROWTH CONDITIONS
- Photoperiod: 16 h
- Light source: Special-fluorescent tube Fluora and Daylight (OSRAM)
- Light intensity and quality: nominal: > 3500 lx, actual: 4954 ± 525
- Day/night temperatures: 22 +/- 10 °C, actual: 13 - 21 °C
- Relative humidity (%): 70 +/- 25, actual: 58 - 100
- Watering regime and schedules: Bottom watering of the test containers with nutrient solution as necessary.


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
During the 21 day observation period the plants were observed every 7 days for number of emerged seedlings, number of dead plants and visual
phytotoxic effects. The rating of the treated plants was done in relation to the control plants. Observations included all variations, either inhibitory
or stimulatory, between the treated test replicates and the untreated control replicates. Such variations were phytotoxic symptoms (chlorosis,
necrosis, wilting), formative effects and growth and development rates. At the end of the study shoot height and shoot fresh weight were measured.
Day -1: Determination of the WHC and the dry weight of the soil
Day 0 (Experimental starting): Application and sowing as specified above.
Every working day: The test containers were bottom watered and fertilised as needed.
Day 7, 14, 21: The replicates were observed for visual phytotoxic effects number of emerged seedlings and number of dead plants.
Test end (day 21): Final shoot heights and shoot fresh weight measurements were done.


- Phytotoxicity rating system (if used):

(1) Alteration of form and appearance (deformation)
(1.1) Curling / twisting / sticking or other deformation of the leafs
(1.2) Change of appearance
(1.3) Shoot deformations
(1.4) Wilting
(1.5) Leaf shedding
(1.6) Shedding of the leaf apex
(1.7) Stunted growth
(1.8) Lacking of cotyledons

(2) Discolouration of the leafs
(2.1) Lighter green (2.2) Darker green
(2.3) Chlorosis (2.3.1) Spots
(2.3.2) Whole leaf
(2.3.3) Veins
(2.3.4) Between veins
(2.3.5) Marginal
(2.4) Loss of colour
(2.5) Discolouration

(3) Alteration of the leaf cells
(3.1) Necrosis (3.1.1) Spots
(3.1.2) Marginal
(3.1.3) Apical
(3.1.4) Between veins
(3.1.5) Whole leaf

(E) Dead plant (Number of dead plants)


Assessment of the Effects

- = no effects observed / normal growth
+/- = slight effects
+ = medium effects
++ = strong effects


VEHICLE CONTROL PERFORMED: no


TEST CONCENTRATIONS

- Range finding study
Results of the Preliminary Test: Inhibition of Shoot Fresh Weight and Emergence on day 21

Test
concentration Inhibition of shoot fresh weight [%] Inhibition of emergence
[%]
[mg/kg DW] Oats Rape Soybean Oats Rape Soybean
10 4 4 -2 5 -6 -7
100 5 18 -2 0 0 -7
1000 8 16 8 5 -11 -7
Nominal and measured concentrations:
nominal:

Oats: 1000 mg/kg DW (dry weight)
Rape, soybean: 1000 - 400 - 160 - 64 - 25.6 mg/kg DW (dry weight)
Reference substance (positive control):
no
Species:
Avena sativa
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:
growth
Remarks:
(Shoot fresh weight, shoot height)
Species:
Brassica napus
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:
growth
Remarks:
(Shoot fresh weight, shoot height)
Species:
Glycine max (G. soja)
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:
growth
Remarks:
(Shoot fresh weight, shoot height)
Details on results:

SEEDLING EMERGENCE

The percent inhibition of shoot height, shoot fresh weight and rate of emergence is given in the tables below. The tabulated results represent
rounded values calculated on the exact raw data.


Oats: Inhibition [%] of Shoot Height, Shoot Fresh Weight and Rate of Emergence

Test concentration [mg/kg DW] Shoot height [cm] Inhibition [%] Shoot fresh weight [mg] Inhibition [%] Rate of emergence [%] Inhibition [%]
Control 40.5 - 1041 - 98 -
1000 41.8 -3 1029 1 98 0


Rape: Inhibition [%] of Shoot Height, Shoot Fresh Weight and Rate of Emergence

Test concentration [mg/kg DW] Shoot height [cm] Inhibition [%] Shoot fresh weight [mg] Inhibition [%] Rate of emergence [%] Inhibition [%]
Control 15.3 - 1628 - 95 -
25.6 14.8 3 1492 8 93 2
64 14.7 4 1471 10 95 0
160 14.7 4 1487 9 93 2
400 14.5 5 1459 10 95 0
1000 14.5 5 1380 15 95 0


Soybean: Inhibition [%] of Shoot Height, Shoot Fresh Weight and Rate of Emergence

Test concentration [mg/kg DW] Shoot height [cm] Inhibition [%] Shoot fresh weight [mg] Inhibition [%] Rate of emergence [%] Inhibition [%]
Control 28.9 - 2199 - 100 -
25.6 27.5 5 2174 1 98 2
64 26.9 7 2141 3 98 2
160 27.0 7 2028 8 100 0
400 27.5 5 2057 6 100 0
1000 28.6 1 2225 -1 100 0

Negative values = promoted growth




Results with reference substance (positive control):
No reference substance tested
Reported statistics and error estimates:
The percent inhibition of biomass growth (shoot height, shoot fresh weight) and rate of emergence were calculated in relation to the control.

One Way Analysis of Variance (ANOVA) was carried out for the determination of statistically significant differences compared to control replicates. When
running a One Way Analysis of Variance a Normality test and an Equal Variance test were done first. P-values for both Normality and Equal Variance test are
0.05. The -value for ANOVA (acceptable probability of incorrectly concluding that there is a difference) is =0.05.

Visual Phytotoxicity on Day 21

 

Oats: Visual Observations and Number of Dead Plants on Day 21

Test
concentration

[mg/kg DW]

Phyto-
toxic
effects

Phytotoxic effects

Replicate

1

2

3

4

5

6

7

8

control

-

-

-

-

-

-

-

-

1000

-

-

-

-

-

-

-

-

 

 

Rape: Visual Observations and Number of Dead Plants on Day 21

Test
concentration

[mg/kg DW]

Phyto-
toxic
effects

Phytotoxic effects

Replicate

1

2

3

4

5

6

7

8

control

-

-

-

-

-

-

-

-

    25.6

-

-

-

-

-

-

-

-

    64

-

-

-

-

-

-

-

-

  160

-

-

-

-

-

-

-

-

  400

-

-

-

-

-

-

-

-

1000

-

-

-

-

-

-

-

-

 

Soybean: Visual Observations and Number of Dead Plants on Day 21

Test
concentration

[mg/kg DW]

Phyto-
toxic
effects

Phytotoxic effects

Replicate

1

2

3

4

5

6

7

8

control

-

-

-

-

-

-

-

-

    25.6

-

-

-

-

-

-

-

-

    64

-

-

-

-

-

-

-

-

  160

-

-

-

-

-

-

-

-

  400

-

-

-

-

-

-

-

-

1000

-

-

-

-

-

-

-

-

- = no effects observed / normal growth

Validity criteria fulfilled:
yes
Conclusions:
After 21 days no test item related phytotoxic effects were noted for all tested plant species at test concentrations up to 1000 mg/kg DW.
No test item related effects > 15 % were observed on the shoot height, the shoot fresh weight and the rate of emergence of all tested plant species.
Therefore the EC50 -Value is > 1000 mg/kg soil DW for shoot height, shoot fresh weight and rate of emergence for all tested plant species.

The NOEC was determined to be 1000 mg/kg DW for shoot height, shoot fresh weight and rate of emergence for all tested plant species. (see table below).

NOEC Values of Shoot Height, Shoot Fresh Weight and Rate of Emergence

NOEC [mg/kg DW]
Species Shoot height Shoot fresh weight Rate of emergence
Oats 1000 1000 1000
Rape 1000 1000 1000
Soybean 1000 1000 1000

Executive summary:

The phytotoxicity of the test item Celitement (batch number: Not specified) to three terrestrial plant species was determined in a seedling emergence test over a period of 21 days, from 2014 -04 -08 to 2014 -04 -29 according to OECD 208 (July 2006) at Dr.U.Noack-Laboratorien,

31157 Sarstedt, Germany.

Test systems were one monocotyledon, oats (Gramineae), and two dicotyledons, rape (Brassicaceae) and soybean (Leguminosae).

The test was conducted for the plant species oats with the limit concentration1000 mg/kg soil dry weight (DW)and for the plant species rape and soybean with the test concentrations of 1000 - 400 - 160 - 64 - 25.6 mg/kg soil dry weight (DW).

The test item was incorporated into the soil in which the seeds were sown afterwards. The test containers were bottom watered with nutrient solution as needed. The toxic effects of the test item were determined on day 7, 14 and 21 by visual observations (number of emerged seedlings and phytotoxic effects) and on day 21 by shoot height and shoot fresh weight determination.

After 21 days no test item related phytotoxic effects were determined for all tested plant species up to the test concentration 1000 mg/kg DW.

 

No inhibitions > 15 % were determined for shoot height, shoot fresh weight and rate of emergence for all tested plant species (see tables below). Therefore the EC50-Value is > 1000 mg/kg soil DW for shoot height, shoot fresh weight and rate of emergence for all tested plant species.

 

The NOEC was determined to be 1000 mg/kg DW for shoot height, shoot fresh weight and rate of emergence for all tested plant species (see table below).

 

Oats: Inhibition [%] of Shoot Height, Shoot Fresh Weight and Rate of Emergence

Test concentration
[mg/kg DW]

Inhibition [%]

Shoot
height

Shoot fresh weight

Rate of emergence

1000

-3

1

0

 

Negative values = promoted growth

 

Rape: Inhibition [%] of Shoot Height, Shoot Fresh Weight and Rate of Emergence

Test concentration
[mg/kg DW]

Inhibition [%]

Shoot
height

Shoot fresh weight

Rate of emergence

  25.6

3

8

2

  64

4

10

0

  160

4

9

2

  400

5

10

0

1000

5

15

0

 

 Soybean: Inhibition [%] of Shoot Height, Shoot Fresh Weight and Rate of Emergence

Test concentration
[mg/kg DW]

Inhibition [%]

Shoot
height

Shoot fresh weight

Rate of emergence

  25.6

5

1

2

  64

7

3

2

  160

7

8

0

  400

5

6

0

1000

1

-1

0

 

 

  NOEC Values of Shoot Height, Shoot Fresh Weight and Rate of Emergence

 

NOEC [mg/kg DW]

Species

Shoot height

Shoot fresh weight

Rate of emergence

Oats

1000

1000

1000

Rape

1000

1000

1000

Soybean

1000

1000

1000

Description of key information

The NOEC was determined to be 1000 mg/kg DW for shoot height, shoot fresh weight and rate of emergence for all tested plant species.

Key value for chemical safety assessment

Short-term EC50 or LC50 for terrestrial plants:
1 000 mg/kg soil dw
Long-term EC10, LC10 or NOEC for terrestrial plants:
1 000 mg/kg soil dw

Additional information

After 21 days no test item related phytotoxic effects were noted for all tested plant species at test concentrations up to 1000 mg/kg DW. No test item related effects > 15 % were observed on the shoot height, the shoot fresh weight and the rate of emergence of all tested plant species. Therefore the EC50 -Value is > 1000 mg/kg soil DW for shoot height, shoot fresh weight and rate of emergence for all tested plant species. The NOEC was determined to be 1000 mg/kg DW for shoot height, shoot fresh weight and rate of emergence for all tested plant species. (see table below). NOEC Values of Shoot Height, Shoot Fresh Weight and Rate of Emergence NOEC [mg/kg DW] were 1000 for all tested plant species.