polymeric zinc propylenebis(dithiocarbamate);propineb (ISO)

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:

2011-05-10 to 2011-06-01

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:

no

GLP compliance:

no

Remarks:

This study was performed according to OECD TG 208. As all validity criteria were met and no deviation from the current guidelines occurred, this study can be considered as high quality (Klimisch Score 1).

Analytical monitoring:

no

Details on sampling:

Visual phytotoxicity ratings (e.g. chlorosis, necrosis) were recorded for the replicates at each application rate on day 7 and 14 after the emergence of 70% of the seeds in the controls. Phytotoxicity was assessed according to EPPO Standard 135 from survived plants. Phytotoxicity is a subjective assessment and was recorded from the living plants at each assessment time. Plants considered as being dead were not rated for phytotoxicity. Dead plants were removed after counting.

The shoot dry weight was determined at the final assessment. The dry weight was determined on a replicate basis, i.e. all plants in one pot were weighed together. The plants of one pot represent one replicate. Plants were cut directly upon the soil surface and put in already preweighed bags. The bags containing the plants were dried at 60oC and reweighed. The weight of each bag was subtracted from the bag containing the dried plant material in order to obtain the shoot dry weight of each replicate. For the statistical analysis and the reporting the weight of one replicate was divided by the number of plants per pot, resulting in a mean weight per shoot value for each pot.

Vehicle:

no

Details on preparation and application of test substrate:

The spray solution was applied once, at test initiation on the soil surface. The blank control spray solution was 200 L deionized water/ha. The test item was dissolved in deionized water and was applied once with 200 L/ha using a spray chamber equipped with an overhead nozzle, with nozzle height set at 30 cm above the sprayed surface. Application speed: 2 km/h; Spray pressure: 4.1 bar.

The sprayer was calibrated beforehand to deliver 200 L/ha ± 10% by spraying glass plates of known weight and area and weighing them immediately afterwards in order to determine the actual amount of water applied. The mean measured application rate was 204 L/ha calculated based on the values from 5 weighed glass plates.

Control: 200 L deionized water/ha for all species
Treatment: 4.62 kg a.s./ha in 200 L water/ha for all species

Species:

Zea mays

Plant group:

Monocotyledonae (monocots)

Details on test organisms:

Historical germination: 100% (germination tests conducted in petri dishes in 2010)

Species:

other: Sorghum sudanense

Plant group:

Monocotyledonae (monocots)

Details on test organisms:

Historical germination: 90% (germination tests conducted in petri dishes in 2010)

Species:

Lolium perenne

Plant group:

Monocotyledonae (monocots)

Details on test organisms:

Historical germination: 85% (germination tests conducted in petri dishes in 2011)

Species:

Allium cepa

Plant group:

Monocotyledonae (monocots)

Details on test organisms:

Historical germination: 95% (germination tests conducted in petri dishes in 2010)

Species:

Lycopersicon esculentum

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

Historical germination: 100% (germination tests conducted in petri dishes in 2011)

Species:

Helianthus annuus

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

Historical germination: 95% (germination tests conducted in petri dishes in 2010)

Species:

Glycine max (G. soja)

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

Historical germination: 90% (germination tests conducted in petri dishes in 2010)

Species:

Cucumis sativus

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

Historical germination: 100% (germination tests conducted in petri dishes in 2011)

Species:

Brassica napus

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

Historical germination: 100% (germination tests conducted in petri dishes in 2011)

Species:

Beta vulgaris

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

Historical germination: 80%* (germination tests conducted in petri dishes in 2010)

Test type:

seedling emergence and seedling growth test

Study type:

laboratory study

Substrate type:

artificial soil

Remarks:

mixture of 90% silt loam soil + 10% washed sand

Limit test:

yes

Total exposure duration:

14 d

Remarks:

The test continued for 14 days following the emergence of 70% of the control seedlings which occurred 4 to 10 days after sowing.

Test temperature:

Regulated to maintain 23 ± 8°C day and 18 ± 8°C night.

pH:

7.30 (CaCl2)

Moisture:

Humidity was regulated to maintain 70 ± 30%.

Watering: Following the initial top watering to facilitate germination of the seed, further irrigation was provided to the pots by bottom watering via 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.

Details on test conditions:

Pots: Commercial plastic flower pots (10.5 cm diameter)
Sowing: Seeds were introduced manually in the soil. The seeds were pressed into the soil surface and completely invisible covered with a layer of the used soil. Soil was top watered immediately to facilitate germination.
Number of Pots: 4 pots per treatment group.
Number of Seeds: The pots were each prepared with 5 seeds. In total 20 seeds per treatment group were tested.
Fertilisation: No supplementary fertilizer was necessary during the study.
Pot placement: The pots were maintained by chance in a block for each species on the glasshouse benches.

Nominal and measured concentrations:

Nominal application rate: 4.62 kg a.s./ha

Reference substance (positive control):

no

Key result

Species:

other: all species

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

> 1.54 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: emergence, seedling survival, visual phytotoxicity, growth and shoot dry weight

Details on results:

In general this study revealed a very low level of phytotoxicity as a result of a soil application of 4.62 kg a.s. Propineb WG 70A W/ha. Emergence was not affected in Brassica napus (Oilseed rape), Cucumis sativus (Cucumber), and Zea mays (Corn). Emergence was increased in Glycine max (Soybean), Helianthus annuus (Sunflower), Lycopersicon esculentum (Tomato), Allium cepa (Onion), Lolium perenne (Ryegrass), and Sorghum sudanense (Sorghum) by 5.3, 11.1, 5.3, 5.6, 20.0 and 11.1%, respectively. Emergence was reduced in Beta vulgaris (Sugar beet) by 11.1%.

Survival was not affected in nine of the tested species. Survival was reduced in Allium cepa (Onion) by 5.3%. There were no observed phytotoxic symptoms in any of the tested plant species. Shoot dry weight was increased in Beta vulgaris (Sugar beet), Brassica napus (Oilseed rape), Cucumis sativus (Cucumber), Allium cepa (Onion), Lolium perenne (Ryegrass) and Zea mays (Corn) by 6.7, 0.2, 4.4, 60.2, 24.6 and 8.2%, respectively. Shoot dry weight was reduced in Glycine max (Soybean), Helianthus annuus (Sunflower), Lycopersicon esculentum (Tomato) and Sorghum sudanense (Sorghum) by 0.9, 1.0, 6.3 and 14.0%, respectively.

Reported statistics and error estimates:

There were no statistically significant effects in any of the tested plant species.

Table 1

Results of single application of 4.62 kg a.s./ha to the ten plant species tested

Species	Emergence (% inhibition)	Survival* (% inhibition)	Phytotoxicity **	Shoot Dry Weight *** (% inhibition)
Beta vulgaris	11.1	0	0	-6.7
Brassica napus	0	0	0	-0.2
Cucumis sativus	0	0	0	-4.4
Glycine max	-5.3	0	0	0.9
Helianthus annuus	-11.1	0	0	1.0
Lycopersicon esculentum	-5.3	0	0	6.3
Allium cepa	-5.6	5.3	0	-60.2
Lolium perenne	-20.0	0	0	-24.6
Sorghum sudanense	-11.1	0	0	14.0
Zea mays	0	0	0	-8.2

* survival is a measure of treated plants that survived at the end of the study and is expressed as an inhibition compared to the untreated control
*** inhibition or reduction is expressed on a per plant basis
- negative values indicate that there was an increase when compared to the untreated control
Bold figures for shoot dry weight are statistically significant (Pairwise Mann-Whitney-U-test, one sided smaller; p ≤ 0.05).

** Desription of the symptoms assessed:
a = chlorosis (yellowing of green shoot tissue)
b = necrosis (brown shoot tissue)
c = bleaching (shoot tissue without pigmentation)
d = wilting (loss of turgor of shoot tissue)
e = leaf deformation (leaf curl, abnormal leaf shape)
f = stunting (plant height reduced with shorter internode length)

**Rating system for recording the severity of phytotoxic symptoms:
0 = no injury or effect
A = slight symptom (s)
B = moderate symptom (s)
C = severe symptom (s)
D = total plant symptom (s)
E = moribund

Validity criteria fulfilled:

yes

Conclusions:

Following a soil surface application of Propineb WG 70A W at 4.62 kg a.s./ha to ten non-target terrestrial plant species. This study is based on the following method: OECD Guideline for the testing of Chemicals, Terrestrial Plant Test OECD 208: Seedling Emergence and Seedling Growth Test, July 2006. This study can be considered valid as the validity criteria of at least 70% emergence and at least 90% survival of the emerged seedlings during the study period were achieved for the untreated controls of all species tested no adverse effects on emergence, seedling survival, visual phytotoxicity, growth and shoot dry weight exceeding 50% effect were observed in this seedling emergence and seedling growth study.

Executive summary:

The purpose of this study was to evaluate potential phytotoxic effects of Propineb on the seedling emergence and growth of ten non-target terrestrial plant species following a pre-emergence application of 4.62 kg a.s./ha (corresponding to 1.54 mg a.s./kg soil d.w.) onto the soil surface. The parameters measured were emergence, survival of the emerged seedlings, visual phytotoxicity, plant growth stage and shoot dry weight.

 

A total of ten species were tested in this seedling emergence and growth test including six dicotyledonous and four monocotyledonous species representing eight plant families. The following species were treated: Beta vulgaris (Sugar beet), Brassica napus (Oilseed rape), Cucumis sativus (Cucumber), Glycine max (Soybean), Helianthus annuus (Sunflower), Lycopersicon esculentum (Tomato), Allium cepa (Onion), Lolium perenne (Ryegrass), Sorghum sudanense (Sorghum) and Zea mays (Corn)

 

Five seeds of each species were sown in 10.5 cm pots in the glasshouse. The soil surface of the pots were treated with 4.62 kg a.s./ha Propineb formulation using a laboratory track sprayer and a water volume rate of 200 L/ha. Each pot (replicate) contained 5 seeds and there were 20 seeds treated i.e. 4 replicates. Control pots were treated with deionised water.

 

This study can be considered valid as the validity criteria of at least 70% emergence and at least 90% survival of the emerged seedlings during the study period were achieved for the untreated controls of all species tested. Following a soil surface application of Propineb WG 70A W at 4.62 kg a.s./ha to ten non-target terrestrial plant species, no adverse effects on emergence, seedling survival, visual phytotoxicity, growth and shoot dry weight exceeding 50% effect were observed in this seedling emergence and seedling growth study.

Reference 2

Endpoint:

toxicity to terrestrial plants: long-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2011-05-10 to 2011-05-31

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 227 (Terrestrial Plant Test: Vegetative Vigour Test)

Version / remarks:

July 2006

Deviations:

no

GLP compliance:

no

Remarks:

This study was performed according to OECD TG 227. As all validity criteria were met and no deviation from the current guidelines occurred, this study can be considered as high quality (Klimisch Score 1).

Details on sampling:

Visual phytotoxicity ratings (e.g. chlorosis, necrosis) were recorded for the replicates at each application rate 7, 14 and 21 days after application. Phytotoxicity was assessed according to EPPO Standard 135 from survived plants. Phytotoxicity is a subjective assessment and was recorded from the living plants at each assessment time. Plants considered as being dead were not rated for phytotoxicity. Dead plants were removed after counting.

The shoot dry weight was determined at the final assessment. The dry weight was determined on a replicate basis, i.e. all plants in one pot were weighed together. The plants of one pot represent one replicate. Plants were cut directly upon the soil surface and put in already preweighed bags. The bags containing the plants were dried at 60oC and reweighed. The weight of each bag was subtracted from the bag containing the dried plant material in order to obtain the shoot dry weight of each replicate. For the statistical analysis and the reporting the weight of one replicate was divided by the number of plants per pot, resulting in a mean weight per shoot value for each pot.

Vehicle:

no

Details on preparation and application of test substrate:

The spray solution was applied once, at test initiation on the leaves and above-ground portions of plants. The blank control spray solution was 200 L deionized water/ha. The test item was dissolved in deionized water and was applied once with 200 L/ha using a spray chamber equipped with an overhead nozzle, with nozzle height set at 30 cm above the target area (highest leaf density). Application speed: 2 km/h; Spray pressure: 4.1 bar.

The sprayer was calibrated beforehand to deliver 200 L/ha ± 10% by spraying glass plates of known weight and area and weighing them immediately afterwards in order to determine the actual amount of water applied. The mean measured application rate was 204 L/ha calculated based on the values from 5 weighed glass plates.

Application Scheme
Control: 200 L deionized water/ha for all species
Treatment: 4.62 kg a.s./ha in 200 L water/ha for all species

Species:

Zea mays

Plant group:

Monocotyledonae (monocots)

Details on test organisms:

Historical germination: 100% (germination tests conducted in petri dishes in 2010)

Species:

other: Sorghum sudanese

Plant group:

Monocotyledonae (monocots)

Details on test organisms:

Historical germination: 90% (germination tests conducted in petri dishes in 2010)

Species:

Lolium perenne

Plant group:

Monocotyledonae (monocots)

Details on test organisms:

Historical germination: 85% (germination tests conducted in petri dishes in 2011)

Species:

Allium cepa

Plant group:

Monocotyledonae (monocots)

Details on test organisms:

Historical germination: 95% (germination tests conducted in petri dishes in 2010)

Species:

Lycopersicon esculentum

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

Historical germination: 100% (germination tests conducted in petri dishes in 2011)

Species:

Helianthus annuus

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

Historical germination: 95% (germination tests conducted in petri dishes in 2010)

Species:

Glycine max (G. soja)

Plant group:

Dicotyledonae (dicots)

Species:

Cucumis sativus

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

Historical germination: 100% (germination tests conducted in petri dishes in 2011)

Species:

Brassica napus

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

Historical germination: 100% (germination tests conducted in petri dishes in 2011)

Species:

Beta vulgaris

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

Historical germination: 80% (germination tests conducted in petri dishes in 2010)

Test type:

vegetative vigour test

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

yes

Total exposure duration:

21 d

Test temperature:

Regulated to maintain 23 ± 8 °C day and 18 ± 8 °C night.

pH:

7.31 (CaCl2)

Moisture:

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.

Details on test conditions:

Test conditions:
Pots: Commercial plastic flower pots (13 cm diameter)

Sowing: Seeds were introduced manually in the soil. The seeds were pressed into the soil surface and completely invisible covered with a layer of the used soil. Soil was top watered immediately to facilitate emergence. To reach the 2-4 leaf stage for application, sowing was started prior to testing.
Number of Pots: 5 pots per treatment group.
Number of Plants: The pots were prepared with 5 - 10 seeds. When more than 4 plants have emerged, they were thinned to 4 plants per pot for application of the test item. In total 20 plants per treatment group were tested.

Fertilisation: For all plants 1 mL Wuxal Universal fertilizer, except Onion with 0.5 mL per pot were added seven days after application. Cucumis sativus was additionally fertilized on test days 1 and 14.

Pot placement: The pots were maintained by chance in a block for each species on the glasshouse benches.

Standard soil (silt loam) from Bayer CropScience AG, Global Biology Herbicides, Horticulture, H 872, Industriepark Hoechst, D-65926 Frankfurt am Main. The soil was sieved to 2 mm. The soil was analysed separately with GLP in the laboratory of LUFA Speyer, Obere Langgasse 40 D-67346 Speyer.

Nominal and measured concentrations:

Nominal application rate: 4.62 kg a.s./ha

Reference substance (positive control):

no

Key result

Species:

other: all species

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

> 1.54 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: shoot dry weight, survival, phytotoxicity

Details on results:

Slight phytotoxic symptoms were observed as chlorosis, necrosis or stunting in Glycine max (Soybean) and Allium cepa (Onion) only in a single pot in the two plant species. Shoot dry weight was increased in Beta vulgaris (Sugar beet), Glycine max (Soybean), Helianthus annus (Sunflower), Lolium perenne (Ryegrass) and Sorghum sudanense (Sorghum) by 16.9, 19.7, 10.4, 16.6 and 71.1%, respectively. Shoot dry weight was reduced in Brassica napus (Oilseed rape), Cucumis sativus (Cucumber), Lycopersicon esculentum (Tomato), Allium cepa (Onion) and Zea mays (Corn) by 3.9, 10.6, 8.3, 22.7 and 6.7%, respectively.

Reported statistics and error estimates:

Statistical analysis of data was performed to obtain significance for shoot dry weight effects, carried out using the Pairwise Mann-Whitney-U-Test (one sided smaller; p = 0.05) by ToxRat statistics. There were no statistically significant effects in any of the tested plant species.

Table 1. Results of single application of 4.62 kg a.s./ha to the 10 plant species tested

Species	Survival* (% inhibition)	Phytotoxicity **	Shoot Dry Weight *** (% inhibition)
Beta vulgaris	0	0	-16.9
Brassica napus	0	0	3.9
Cucumis sativus	0	0	10.6
Glycine max	0	0 - A ab	-19.7
Helianthus annuus	0	0	-10.4
Lycopersicon esculentum	0	0	8.3
Allium cepa	0	0 - A af	22.7
Lolium perenne	0	0	-16.6
Sorghum sudanense	0	0	-71.1
Zea mays	0	0	6.7

* survival is a measure of treated plants that survived at the end of the study and is expressed as an inhibition compared to the untreated control
*** inhibition or reduction is expressed on a per plant basis
- negative values indicate that there was an increase when compared to the untreated control
Bold figures for shoot dry weight are statistically significant (Pairwise Mann-Whitney-U-test, one sided smaller; p ≤ 0.05).

** Desription of the symptoms assessed:
a = chlorosis (yellowing of green shoot tissue)
b = necrosis (brown shoot tissue)
c = bleaching (shoot tissue without pigmentation)
d = wilting (loss of turgor of shoot tissue)
e = leaf deformation (leaf curl, abnormal leaf shape)
f = stunting (plant height reduced with shorter internode length)

**Rating system for recording the severity of phytotoxic symptoms:
0 = no injury or effect
A = slight symptom (s)
B = moderate symptom (s)
C = severe symptom (s)
D = total plant symptom (s)
E = moribund

Validity criteria fulfilled:

yes

Conclusions:

This study is based on the following method: OECD Guideline for the testing of Chemicals,
Terrestrial Plant Test OECD 227: Vegetative Vigour Test, July 2006. The purpose of this specific study was to evaluate potential phytotoxic effects of Propineb formulation on the vegetative vigour of ten non-target terrestrial plant species following a postemergence application of 4.62 kg a.s./ha onto the foliage of plants at the 2-4 leaf stage. This study can be considered valid as the validity criterion of at least 90% survival throughout
the study period was achieved for the untreated controls of all species tested. No adverse effect on survival, visual phytotoxicity, growth and shoot dry weight was observed after foliar application of 4.62 kg a.s./ha of propineb. Growth and shoot dry weight exceeding 50% effect were observed in this vegetative vigour study.

Executive summary:

The purpose of this specific study was to evaluate the potential phytotoxic effects of Propineb on the vegetative vigour of ten non-target terrestrial plant species following a post emergence application of 4.62 kg a.s./ha the foliage of plants at the 2-4 leaf stage. This study is based on the following method: OECD Guideline for the testing of Chemicals, Terrestrial Plant Test OECD 227: Vegetative Vigour Test, July 2006. At the 2-4 leaf stage plants were treated with 4.62 kg a.s./ha Propineb WG 70A W using a laboratory track sprayer and a water volume rate of 200 L/ha. Each pot (replicate) contained 4 plants and there were 20 plants treated i.e. 5 replicates. Control pots were treated with deionised water. This study can be considered valid as the validity criterion of at least 90% survival throughout the study period was achieved for the untreated controls of all species tested.

Following a foliar application of Propineb at 4.62 kg a.s./ha (corresponding to 1.54 mg a.s./kg soil d.w.) to ten non-target terrestrial plant species at the 2 to 4 leaf stage, no adverse effects on survival, visual phytotoxicity, growth and shoot dry weight exceeding 50% effect were observed in this vegetative vigour study.

Description of key information

OECD Guideline for the testing of Chemicals, Terrestrial Plant Test OECD 227: Vegetative Vigour Test, July 2006 and OECD Guideline for the testing of Chemicals, Terrestrial Plant Test OECD 208: Seedling Emergence and Seedling Growth Test, July 2006 were followed.

Following a foliar application of Propineb at 4.62 kg a.s./ha (corresponding to 1.54 mg a.s./kg soil d.w.) to ten non-target terrestrial plant species at the 2 to 4 leaf stage, no adverse effects on survival, visual phytotoxicity, growth and shoot dry weight exceeding 50% effect were observed in this vegetative vigour study.

Following a soil surface application of Propineb at 4.62 kg a.s./ha (corresponding to 1.54 mg a.s./kg soil d.w.) to ten non-target terrestrial plant species, no adverse effects on emergence, seedling survival, visual phytotoxicity, growth and shoot dry weight exceeding 50% effect were observed in this seedling emergence and seedling growth study.

Key value for chemical safety assessment

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

1.54 mg/kg soil dw