polymeric zinc propylenebis(dithiocarbamate);propineb (ISO)

Toxicity to terrestrial arthropods

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

toxicity to terrestrial arthropods: short-term

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

2011/08/16 to 2011/11/14

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 226 (Predatory Mite (Hypoaspis (Geolaelaps) Aculeifer) Reproduction Test in Soil)

Version / remarks:

2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Application method:

mixed into soil

Analytical monitoring:

no

Details on sampling:

1st test run: 10 adult soil mites (females) were exposed to 100, 178, 316, 562 and 1000 mg product/kg dry weight. They were fed every 2 days with Tyrophagus putrescentiae (SCHRANK). Mortality and reproduction were determined after 14 days of exposure.

2nd test run: 10 adult soil mites (females) were exposed to 5.6, 10, 18, 32 and 56 mg product/kg dry weight. They were fed every 2 days with Tyrophagus putrescentiae (SCHRANK). Mortality and reproduction were determined after 14 days of exposure.

Vehicle:

no

Details on preparation and application of test substrate:

The composition of the soil was 5% sphagnum peat; origin: classified according to DIN 11540 (as close to pH 5.5-6.0 as possible, no visible plant remained, finely ground, dried to measured moisture content). 20% kaolin clay (kaolinite content > 30%); type: Kaolin W, - 0.2% calcium carbonate; - 74.8% industrial quartz sand; type: Millisil W3, (fine sand is dominant with more than 50% of the particles between 50 and 200 µm) and deionised water.

The amount of soil per treatment group: 200 g soil dry weight, the amount of soil/test vessel: 20 g soil dry weight (height of soil approximately 1.6 cm). The Max. water holding capacity WHC (g/100 g soil d.w.): 39.79 and the water content on the 1st test run: test termination: 18.57 - 19.57% (equivalent to 46.67 - 49.19% of WHC) test start: 19.81 - 20.14% (equivalent to 49.78 - 50.61% of WHC). For the 2nd test run: test termination: 18.65 - 19.63% (equivalent to 46.87 - 49.34% of WHC) test start: 19.64 - 20.12% (equivalent to 49.35 - 50.55% of WHC).

The pH value for the 1st test run: test termination: 5.4 - 5.5 test start: 5.5 - 5.7. For the 2nd test run: test termination: 5.4 - 5.8 test start: 5.6 - 5.7.

Test organisms (species):

Hypoaspis aculeifer

Animal group:

other: Canestrini

Details on test organisms:

Reared in test facility under ambient conditions, the breeding medium was a mixture of plater of Paris, activated charcoal and deionised water (8:1:9). The breeding units were passively ventilated plastic vessels (12 cm in diameter and 8cm in height) with a layer of carbon darkened plaster of Paris. The specimens were fed Tyrophagus Putrescentiae (Schrank) supplied twice to three times a week and the age of test organisms were adult from a synchronized culture with an age difference of 3 days.

Study type:

laboratory study

Limit test:

no

Total exposure duration:

14 d

Test temperature:

guideline requirement: 20 ± 2 °C
1st test run:
19.5 - 21.2 °C
2nd test run:
19.5 - 21.1 °C

pH (if soil or dung study):

1st test run:
test termination: 5.4 - 5.5
test start: 5.5 - 5.7
. 2nd test run:
test termination: 5.4 - 5.8
test start: 5.6 - 5.7

Photoperiod and lighting:

light : dark cycle = 16 h : 8 h (artificial light; 494 - 496 lx)

Details on test conditions:

The lighting source was artificial light (Lumilux L58W), with a: guideline intensity requirement of 400 - 800 lx. The 1st test run was 621 lx, the 2nd test run: duration: 493 lx and the light : dark duration was 16 h : 8 h. The test vessels were 100ml Schott-bottles with screw cap (4 cm in diameter and 11 cm in height ). There were 10 organisms per vessel, 4 replicates in treated groups (+ 2 replicates for determination of water content and pH value), 8 replicates in control groups (+ 2 replicates for determination of water content and pH-value; not loaded with predatory mites). 80 predatory mites in control group and 40 in treated group.

At day 29 (1st test run) or 35 (2nd test run) after transfer of the parental females to the rearing vessels for egg-laying, the mites of the synchronized cultures were suitable for the respective tests. At test start, adult females of the synchronized culture were transferred to the prepared test vessels which contained untreated (control) or test item treated artificial soil (20 g soil dry weight) with a water content of 40-60 % of the maximum water holding capacity (WHC). Per test vessel 10 adult females were introduced by means of a moistened brush (= start of exposure).

Nominal and measured concentrations:

1st test run: 100, 178, 316, 562, 1000 mg product/kg soil dry weight
2nd test run: 5.6, 10, 18, 32, 56 mg product/kg soil dry weight

Reference substance (positive control):

yes

Remarks:

Dimethoate EC 400

Key result

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

39.1 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Key result

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

392 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Key result

Duration:

14 d

Dose descriptor:

LOEC

Effect conc.:

698 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Key result

Duration:

14 d

Dose descriptor:

LOEC

Effect conc.:

69.8 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Key result

Duration:

14 d

Dose descriptor:

LC50

Effect conc.:

> 698 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Key result

Duration:

14 d

Dose descriptor:

EC50

Effect conc.:

> 39.1 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Details on results:

Study was performed with the formulated product Propineb WG 70A W.

In the control group, the mortality rate was 2.5% and 3.8% in the 1st and 2nd test run, respectively. Mortality rates of 0.0 - 30.0% were recorded in the treatment groups of both test runs. The observed mortality rates for adult mortality in the treatment groups compared to control were not statistically significant up to and including a test concentration of 562 mg product/kg soil d.w. (Fisher’s Exact Binomial Test, a = 0.05, one-sided greater). However, a statistically significant effect on mortality compared to control was found at 1000 mg product/kg soil d.w.

Fourteen days after introduction of the parental mites into the test vessels of the 1st test run, the mean number of juveniles was 271.8 in the control and 110.0, 0.0, 6.0, 0.5 and 0.0 at the test concentrations of 100, 187, 316, 562 and 1000 mg product/kg soil d.w., respectively. The mean number of juveniles of the 2nd test run was 273.5 in the control and 275.0, 278.3, 293.0, 274.3 and 241.3 at the test concentrations of 5.6, 10, 18, 32 and 56 mg product /kg soil d.w., respectively.

Results with reference substance (positive control):

The EC50 (reproduction) of the reference item Dimethoate EC 400 was calculated to be 5.4 mg/kg soil d.w..

Reported statistics and error estimates:

Mortality (number of dead adults) in % for the treatment groups was calculated. Missing mites were counted as dead. Observations in the treatment group were expressed relative to the water control group. The reduction of reproductive for each treatment group was calculated in % compared to control. The statistical analysis was performed with the software ToxRat Professional 2.10.05 (RATTE 2010). Fisher’s Exact Binomial Test, Welch t-test and Dunnett-t-test were used to compare the control with the independent test item groups.

No statistically significant reduction of reproduction up to and including a test concentration of 56 mg product/kg soil dry weight (Dunnett-t-test, a = 0.05, one-sided smaller). However, statistically significant effects on reproduction compared to control were found beginning at a concentration of 100 mg product/kg soil d.w. (Welch t-test, a = 0.05, one-sided smaller). No statistically significant reduction of reproduction up to and including a test
concentration of 56 mg product/kg soil dry weight (Dunnett-t-test, a = 0.05, one-sided smaller). However, statistically significant effects on reproduction compared to control were found beginning at a concentration of 100 mg product/kg soil d.w. (Welch t-test, a = 0.05, one-sided smaller).

Table one

Effects of the test item on Hypoaspis aculeifer mortality and reproduction first test run

Endpoint	Propineb (mg product/kg soil d.w)
	control	100	178	316	562	1000
Mortality of soil mites after 14 days (%)	2.5	7.5	0.0	5.0	2.5	30.0*
Mean number of juveniles after 14 days	271.8	110.0*	0.0*	6.0*	0.5*	0.0*
CV (%)	15.7	112.5	- ^1)	127.7	115.5	- ^1)
Reduction of reproduction (% to control)	-	59.5	100.0	97.8	99.8	100.0

-1) not determined due to mathematical reasons
* statistically significant difference compared to control
(Fisher`s Exact Binomial Test for mortality, α = 0.05; Welch t-test for reproduction; α = 0.05)
Calculations were done using non-rounded values
Percent reduction: (1 - Rt / Rc) * 100 %
Rt = mean number of juvenile mites observed in the treated group(s)
Rc = mean number of juvenile mites observed in the control group
Negative values = increase, relative to control

 

 

Table two

 

Effects of the test item on Hypoaspis aculeifer mortality and reproduction second test run

Endpoint	Propineb  (mg product/kg soil d.w)
	control	5.6	10	18	32	56
Mortality of soil mites after 14 days (%)	3.8	2.5	2.5	0.0	7.5	0.0
Mean number of juveniles after 14 days	273.5	275.0	278.3	293.0	274.3	241.3
CV (%)	7.9	15.5	^6.7	7.7	2.5	^17.2
Reduction of reproduction (% to control)	-	-0.5	-1.7	-7.1	-0.3	11.8

No statistically significant differences compared to the control were calculated (Fisher`s Exact Binomial Test for mortality, α = 0.05;
Dunnett-t-test for reproduction; α = 0.05)
Calculations were done using non-rounded values
Percent reduction: (1 - Rt / Rc) * 100 %
Rt = mean number of juvenile mites observed in the treated group(s)
Rc = mean number of juvenile mites observed in the control group
Negative values = increase, relative to control

Validity criteria fulfilled:

yes

Conclusions:

The purpose of this study was to determine potential effects of the Propineb WG 70A W on the mortality and the reproductive output of the soil mite species Hypoaspis aculeifer (CANESTRINI) during a test period of 14 days according to the OECD guideline 226 (2008). Concentrations of 100, 178, 316, 562, 1000 mg product/kg dry weight soil was tested in the 1st run and 5.6, 10, 18, 32 and 56 mg product/kg dry weight soil in the 2nd run. The NOEC for mortality and reproduction were determined to be 392 and 39.1 mg a.s./kg soil dry weight, respectively. The LOEC for mortality and reproduction were determined to be 698 and 69.8 mg a.s./kg soil dry weight, respectively.

Executive summary:

The purpose of this study was to determine potential effects of the Propineb on the mortality and the reproductive output of the soil mite species Hypoaspis aculeifer (CANESTRINI) during a test period of 14 days according to the OECD guideline 226 (2008). Concentrations of 100, 178, 316, 562, 1000 mg product/kg dry weight soil was tested in the 1^st run and 5.6, 10, 18, 32 and 56 mg product/kg dry weight soil in the 2^nd run. The NOEC for mortality and reproduction were determined to be 392 and 39.1 mg a.s./kg soil dry weight, respectively. The LOEC for mortality and reproduction were determined to be 698 and 69.8 mg a.s./kg soil dry weight, respectively.

Reference 2

Endpoint:

toxicity to terrestrial arthropods: short-term

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

2011/08/02 to 2011/08/30

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

ISO 11267 (Inhibition of Reproduction of Collembola by Soil Pollutants)

Version / remarks:

1999

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 232 (Collembolan Reproduction Test in Soil)

Version / remarks:

2009

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Application method:

mixed into soil

Details on sampling:

The test was started using juvenile collembolans, Folsomia candida, 9 - 12 days old. Test organisms of a uniform age were obtained by transferring egg clusters from the breeding containers to fresh containers with fresh substrate 12 days before starting the experiment. After 72 hours these egg clusters were removed from the containers and the juveniles that had hatched during the preceding 72 hours were fed granulated dry yeast. After a further 9 days the test organisms were collected and used for the test. Ten test organisms were introduced to each vessel, using an exhauster. After addition of the test organisms the test vessels were positioned randomly in a controlled-environment test room, and these positions were re-randomized weekly. The test containers were tightly covered with a glass lid and briefly opened twice a week for aeration. The test organisms were fed twice during the experiment (at the start of the test and after 14 days) with approximately 2 mg of granulated dry yeast per test vessel.

Four weeks after introducing the test organisms the parental and juvenile collembolans in the test and control vessels were counted. The test substrate of each replicate was poured into an individual 150 - 200 mL container and the test organisms floated off the substrate by the addition of water. To improve the contrast between the white collembolans and surrounding water surface, the water was stained dark with ink. After gentle stirring the numbers of parental and juvenile collembolans floating on the surface were determined. Missing parental collembolans are assumed to have died during the test period. Surviving adults and juveniles were counted using a digital image processing system (LemnaTec Scanalyzer), an automated counting technique based on a video camera connected to a digital image storage and analysis system. This technique fulfils the requirement of the ISO guideline regarding precision of the counting method (average error <10 %).

Vehicle:

no

Details on preparation and application of test substrate:

Two days before the start of the test dry artificial soil was pre-moistened by adding deionised water to obtain approximately half of the final water content. On the day of the test start the test item was introduced by dispersing the quantity of test item required to obtain the desired test concentration in the volume of water required to hydrate the soil to 40-60 % of its WHC. The control substrate contained the corresponding amount of deionised water only. After thorough mixing, 30 g (wet weight) of the test substrate was placed into each vessel, avoiding compression.

An exactly weighed amount of the test item was dispersed in deionised water to make a stock, without addition of solubility mediators, immediately before application. This stock solution was diluted with deionised water such that each dispersion contained the amount of test item in 25 mL required to dose 250 g soil dry weight (at 15 % dry weight water content). Each test item dispersion (25 mL) was thoroughly mixed with the artificial soil using a laboratory mixer, achieving a final water content of 40-60 % of WHC.

Composition of soil: was 5 % sphagnum peat; classified according to DIN 11540 (as close to pH 5.5-6.0 as possible, no visible plant remained, finely ground, dried to measured moisture content). 20 % kaolin clay (kaolinite content > 30 %); type: Kaolin W, 0.3 % calcium carbonate; 74.7 % industrial quartz sand; type: Millisil W3, (fine sand is dominant with more than 50 % of the particles between 50 and 200 µm) and deionised water.

Test organisms (species):

Folsomia candida

Animal group:

Collembola (soil-dwelling springtail)

Details on test organisms:

The replicas were reared in the laboratory of the test facility under ambient laboratory conditions.

The breeding container was a plastic vessel of Bellaplast with a transparent lid (9.5 cm x 9.5 cm x 6 cm) moistening automatically by means of an absorbent wick aeration in combination with feeding food source: granulated dry yeast, supplied twice a week light : dark cycle = 16 h : 8 h (diffuse artificial light) temperature: nominally 20±2 °C.

The juvenile collembolans were between 9-12 days old and were fed granulated dry yeast.

Study type:

laboratory study

Limit test:

no

Total exposure duration:

28 d

Test temperature:

18.0 – 22.0 °C

pH (if soil or dung study):

5.67 – 5.71

Photoperiod and lighting:

source: artificial light (Lumilux L58W)
intensity: 720 lx
duration: light : dark = 16 h : 8 h

Details on test conditions:

Test environment was a climatic chamber.
Test vessel was a glass container (approximately 150 mL) covered with a glass lid
Number of collembolans per vessel was (= replicate): 10
Number of replicates in the control group: 8 ( + 2 replicates not loaded with collembolans for measurement purposes).
Number of replicates in the treated group: 4 ( + 2 replicates not loaded with collembolans for measurement purposes).
Number of collembolans in the control group was 80
Number of collembolans in the treated group of 40 individuals
Feeding regime was approximately 2 mg granulated dry yeast at the start of the test and after 14 days

Nominal and measured concentrations:

18, 32, 56, 100 mg test item/kg soil dry weight

Reference substance (positive control):

yes

Remarks:

The reference item boric acid (100 % analysed) was tested in a separate study at concentrations of 44, 67, 100, 150 and 225 mg/kg soil dry weight

Toxic reference:

To verify the sensitivity of the test system the reference item boric acid is routinely tested at concentrations of 44, 67, 100, 150 and 225 mg/kg soil dry weight. The collembolans of the reference test were from the same source culture as those used in the definitive test. In a most recent study the EC50 was determined to be 107 mg/kg soil dry weight. The LC50 was determined to be 193 mg/kg soil dry weight. The NOEC for mortality and for reproduction was determined to be 67 and 44 mg/kg soil dry weight, respectively. The EC50 value for the reduction of reproduction was close to the value of 100 mg/kg soil dry weight as stated in OECD 232 (2009). The EC50 therefore showed that the test system was sensitive.

Key result

Duration:

28 d

Dose descriptor:

LOEC

Effect conc.:

69.8 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks:

Adults

Key result

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

39.1 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks:

Adult

Key result

Duration:

28 d

Dose descriptor:

LC50

Effect conc.:

89.3 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks:

Adult

Key result

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

39.1 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Key result

Duration:

28 d

Dose descriptor:

LOEC

Effect conc.:

69.8 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Key result

Duration:

28 d

Dose descriptor:

EC50

Effect conc.:

67 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Details on results:

Study was conducted with the formulated product Propineb WG 70A W.

Mortality rates of 5.0% - 45.0% were recorded in the test item treatment groups. In the control the mortality rate was 6.3%. Statistically significant effects compared to the control were observed at a concentration of 100 mg form./kg soil d.w. (Fisher`s Exact Binomial Test, alpha = 0.05, one-sided greater).
Statistically significant effects (Welch-t-test, alpha = 0.05, one-sided smaller) on the number of juveniles compared to the control group were recorded at a concentration of 100 mg form./kg soil d.w. The mean reproduction in the control reached 436 juveniles. Reproduction rates in the concentrations of 10, 18, 32, 56 and 100 mg form./kg soil d.w. were 421, 434, 429, 431 and 185 juveniles, respectively.

Reported statistics and error estimates:

The statistical analysis was performed with the software ToxRat Professional 2.10.05 (RATTE 2010). Fisher`s Exact Binomial Test with Bonferroni Correction and the Welch-t-test for Inhomogeneous Variances with Bonferroni Adjustment were used to compare the control with the independent test item groups. The LC50 and EC50 were calculated by Probit analysis using linear weighted regression and Probit analysis using the linear maximum likelihood regression, respectively (Finney 1971).

Table two

Effects of Propineb on mortality of parental collembolans

 

Treatment group	control	10 (mg form./kg soil dw)	18 (mg form./kg soil dw)	32 (mg form./kg soil dw)	56 (mg form./kg soil dw)	100 (mg form./kg soil dw)
Replicate	Number of survivng parental collembolans per replicate (4 weeks after test initiation)
1 2 3 4 5 6 7 8	9 10 8 10 10 9 9 10	10 10 9 9	9 9 9 10	9 8 10 10	10 8 10 9	7 4 6 5
Mean ±SD (cv %)	9.4  0.7 7.9	9.5 0.6 6.1	9.3 0.5 5.4	9.3 1.0 10.4	9.3 1.0 10.4	5.5 * 1.3 23.5
Mortality (%)	6.3	5.0	7.5	7.5	7.5	45.0

* statistically significantly different from control
(Fisher’s Exact Binomial Test with Bonferroni Correction, α = 0.05, one-sided greater)
SD: standard deviation, cv %: coefficient of variation, d.w.: dry weight (of artificial soil)

 

 

Table three

 

Treatment group	control	10 (mg form./kg soil dw)	18 (mg form./kg soil dw)	32 (mg form./kg soil dw)	56 (mg form./kg soil dw)	100 (mg form./kg soil dw)
Replicate	Number of juveniles per replicate (4 weeks after test initiation)
1 2 3 4 5 6 7 8	341 474 427 423 495 435 427 465	455 392 489 347	359 328 470 578	423 404 442 445	455 444 408 418	240 147 231 123
Mean ±SD (cv %)	436 46.5 10.7	421 63.5 15.1	434 113.9 26.3	429 19.0 4.4	431 21.9 5.1	185 * 59.0 31.8
% Reproduction of reproduction compared to control	-	3	0	2	1	58

* statistically significantly different from control (Welch-t-test for Inhomogeneous Variances with Bonferroni
Adjustment for reproduction; alpha = 0.05, one-sided smaller)
SD: standard deviation, cv %: coefficient of variation, d.w.: dry weight (of artificial soil)
calculations were done using unrounded values
Percent reduction: (1-Rt/Rc) * 100 %
Rt = mean number of juveniles observed in the treated groups
Rc = mean number of juveniles observed in the control group
Negative values indicate a higher reproductive performance compared to control.

 

Table four 

 

	Propineb WG 70A W (mg form./kg soil dry weight)
	Mortality	Reproduction
LOEC LC10 / EC10 LC20 / EC20 LC50 / EC50 (95% confidence limit, lower-upper) NOEC	100 51 69 128 (92-178)   56	100 71 79 96 (92-98)   56

Validity criteria fulfilled:

yes

Remarks:

Mean adult mortality = 20 % (observed: 6.3 %) Mean number of juveniles per test vessel = 100 (observed: average of 436/vessel) Coefficient of variation for the mean number of juveniles: < 30 % (observed: 10.7 %)

Conclusions:

The purpose of this study is to determine potential effects of different concentrations of the formulation on the reproductive output of the collembolans Folsomia candida during a test period of 28 days. After 4 weeks the number of offspring (juveniles) and surviving parental collembolans were counted. Collembolans were exposed to 10, 18, 32, 56 and 100 mg form./kg soil dry weight under OECD guideline 232 (2009) and ISO guideline 11267 (1999).
The formulation caused a significant reduction of reproduction of the collembolans at 69.8 mg a.s./kg soil dry weight, i.e. the highest concentration tested. Therefore, the overall No-Observed-Effect- Concentration (NOEC) was determined to be 39.1 mg a.s./kg soil d.w., and the Lowest-Observed-Effect- Concentration (LOEC) was determined to be 69.8 mg a.s./kg soil d.w.

Executive summary:

The purpose of this study is to determine potential effects of different concentrations of the formulation on the reproductive output of the collembolans Folsomia candida during a test period of 28 days. After 4 weeks the number of offspring (juveniles) and surviving parental collembolans were counted. The test was performed in accordance with the OECD Guideline 232 (2009) and the International Standard ISO 11267 (1999).

The overall NOEC (mortality and reproduction) was determined to be 39.1 mg a.s./kg soil d.w..

Description of key information

2 studies of different terrestrial arthropods summaries are documented below.

M-416315-01-1, Friedrich, 2011

The purpose of this study is to determine potential effects of different concentrations of the propineb on the reproductive output of the collembolans Folsomia candida during a test period of 28 days.

The overall NOEC was determined to be 39.1 mg a.s./kg soil d.w..

 

M-421441-01-1, Schulz, 2011

The purpose of this study was to determine potential effects of the Propineb WG 70A W on the mortality and the reproductive output of the soil mite species Hypoaspis aculeifer (CANESTRINI) during a test period of 14 days according to the OECD guideline 226 (2008). Concentrations of 100, 178, 316, 562, 1000 mg product/kg dry weight soil was tested in the 1^st run and 5.6, 10, 18, 32 and 56 mg product/kg dry weight soil in the 2^nd run. The NOEC for reproduction was determined to be 39.1 mg a.s./kg soil dry weight.

 

Additional 4 studies on bees available and executive summaries are documented below.

M-442120-01-1, Schmitzer, 2012

The acute oral toxicity and acute contact toxicity of propineb was assessed in a limit test in Apis mellifera. Under laboratory conditions Apis mellifera 50 worker bees per treatment were exposed for 48 hours to a single dose of 100.0 µg a.s. per bee for topical application (contact limit test) and to a single dose of 107.9 µg a.s. per bee for feeding (oral limit test, value based on the actual intake of the test item).

The results can be considered as valid, as all validity criteria of the test were met: control mortality is <10% in the oral and in the contact test, LD₅₀ (24 h) of the toxic standard in the oral test equals 0.16 mg a.s./bee, the LD₅₀ (24 h) of the toxic standard in the contact test equals 0.17 mg/bee. LD₅₀ oral (solution in acetone and mixed with syrup) >107.9 µg a.s./bee (0% mortality after 48h), no behavioral effects were observed at any time in the oral toxicity test. LD₅₀ contact (solution in acetone) >100.0 µg a.s./bee (8% mortality after 48h), one bee was apathetic during the 4 hours and 48 hours assessment.

M-454682-01-1, Schmitzer, 2013

This study was designed to assess the effects of test substance to the honey bee brood. The employed method of investigating the development of the honey bee brood is based on the method of Oomen et al. (1992).

Ontogenesis of eggs, young and old larvae of honey bees were observed. Mortality of the honey bees and sublethal effects, such as changes in behaviour, were also monitored. The results were compared to an untreated control and to a reference item (fenoxycarb). The mixing ratio was 3.07 g Propineb WG 70A W in 1 L sugar syrup. Also bee brood development (eggs, young and old larvae) were recorded one day before test initiation and after 4, 8, 15 and 22 days. As control pure sugar syrup (30% sucrose, 31% glucose, 39% fructose) was used. 3.0 g/L syrup Insegar (25% fenoxycarb, 0.75 g fenoxycarb/L) was used as reference substance

The mean termination rate of eggs after treatment with Propineb formulation was statistically significantly higher (38.4%) when compared to the values of the control group (16.0%). NOED for young and old larvae and adult bee mortality was determined to be 2.10 g a.s./L.

M-017002-01-1, Kleiner, 1998

The acute oral toxicity and acute contact toxicity of propineb was assessed in a limit test to Apis mellifera.

Exposure to propineb caused negligible mortality (2% and 4%, respectively) up to a dose of 200 µg a.s./bee in the oral and the contact toxicity test. Therefore, it is concluded that the LD₅₀ (48 h) is >200 µg a.s./bee in the oral and the contact toxicity test.

No effects on the behaviour of the bees (or other sublethal effects) were observed in comparison with the control bees.

M-487104-01-1, Kling, 2014

The purpose of this study was to determine the chronic effects of the test item Propineb formulation on the honey bee, Apis mellifera L., in a 10 days continuous feeding test in the laboratory. Honey bees were exposed to a 50% aqueous sucrose application (feeding) solution containing the nominal concentration level of 120 mg a.s./kg of Propineb formulation by continuous and ad libitum feeding over a period of 10 days.

Mortality and sublethal effects were observed every day at about the same time of day. The chronic effects of the test item Propineb formulation as well as the NOEC value were evaluated by comparing the results of the test item treatment to those of the control treatment.

The NOEC for mortality was determined at the end of the test period to be 120 mg a.s./kg (nominal). The LC₅₀ was determined to be >120 mg a.s./kg (nominal). At 120 mg a.s./kg, no remarkable sub-lethal effects or behavioural abnormalities were observed throughout the entire observation period of 10 days.