polymeric zinc propylenebis(dithiocarbamate);propineb (ISO)

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2004/06/01 to 2004/07/21

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: JMAFF 12 Nousan No. 8147

Version / remarks:

2000

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1010 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids)

Version / remarks:

1996

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPP 72-2 (Aquatic Invertebrate Acute Toxicity Test)

Version / remarks:

1982

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

1984 and corresponding revised draft proposal (2004)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Duplicate samples containing 20 mL of the freshly prepared test media have been taken at test initiation from all test levels and the controls prior to introducing the test animals. For additional stability measurements the aged replicates of each concentration level were pooled after 2 days in one vessel and samples have then be taken from this pooled media.
Samples, not intended for immediately measurement, have been stored deep frozen.

Vehicle:

yes

Remarks:

Dimethylformamide (DMF)

Details on test solutions:

Initially a stock solution (stock solution A) was prepared by dissolving 2915.8 mg test item in 50 mL DMF (Dimethylformamide) (= 48g a.s./L). This solution was ultrasonified for 5 minutes, followed by 26 minutes of stirring.

From this solution, sub-dilutions were established:
Stock solution B >5 mL solution A + 5 mL DMF = 24 g test item/L
Stock solution C >5 mL solution B + 5 mL DMF = 12 g test item/L
Stock solution D > mL solution C + 5 mL DMF = 6.0 g test item/L
Stock solution E >5 mL solution D + 5 mL DMF = 3.0 g test item/L
Stock solution F >5 mL solution E + 5 mL DMF = 1.5 g test item/L

Each solution was stirred until visible homogeneity was achieved.
From each of these stock solutions 100 µL were taken and carefully added to an amount of 1000 mL test water in order to establish the exposure-concentrations.

Test organisms (species):

Daphnia magna

Details on test organisms:

First instars (Daphnia magna) < 24 hours old, from laboratory stock breeding, were used for this study. The first instars used in the test were obtained by repeated careful screening of a breeding culture of defined age, using plastic screens with 0.6 and 0.2 mm mesh size less than 24 h before test initiation. The parent daphnids were of the same age (±12 hours), 21 days old and from the fourth brood or later. No males, ephippia or dead animals were present in the cultures. The cultures showed no delay in first brood.

6 x 5 animals per concentration

Test type:

static

Water media type:

other: "M7 medium" as documented in the "Original Draft" of an EEC Daphnia magna Pilot Ring Test. The medium is prepared using deionised water by adding mineral salts. Immediately before use in a study, the vitamin components will be added.

Limit test:

no

Total exposure duration:

48 h

Remarks on exposure duration:

After 24 and 48 hours, behaviour of the water fleas was visually evaluated.

Hardness:

10° dH (= 178 mg/L CaCO3)

Test temperature:

20.3 - 20.6°C

pH:

7.9 - 8.1

Dissolved oxygen:

8.6 - 8.8 O2 Conc. (mg/L)

Conductivity:

576 µS/cm

Nominal and measured concentrations:

Nominal concentrations: 0.15, 0.30, 0.60, 1.20, 2.40 and 4.80 mg a.s./L, Mean measured concentrations: 0.152, 0.241, 0.498, 1.07, 2.18 and 4.04 mg a.s./L

Details on test conditions:

The water flea Daphnia magna was exposed to Propineb over a 48 hours period under static conditions. The test vessels consisted of 100 mL glass beakers, individually labelled and filled with 50 mL of the test solution, corresponding to a fluid level of approximately 3 cm height. Six vessels (replicates), each provided with five daphnids (= 10 mL test solution per daphnid), were used per treatment group and control (=30 animals per study group). The beakers were covered with transparent glass plates and placed in a climate controlled environment (isolated chamber) between 18 and 22°C (maximum allowed deviation ± 1°C within 48 hours). Lighting was adjusted to a 16:8 hours light dark cycle at about 1400 lux.

The water fleas were not fed and the test solutions were not artificially aerated during exposure. Any surface in contact with the test solution was made of glass or other chemical inert material.

Reference substance (positive control):

no

Key result

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

5.66 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

1.5 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Based on mean-measured concentrations of propineb, the EC50 for immobilisation after 48 hours of static exposure was 1.50 mg a.s./L (95% confidence limits 1.28 -1.76 mg a.s./L).

The test was performed wit the formulated product Propineb VM 80.

Table 2

Toxicity to Daphnia magna (based on nominal concentrations of pure Propineb):

Test concentration			Exposed Daphnids (=100%)	Immobilised Daphnids
mg product/L	mg a.s./L	mg a.s./L		24h		48h
Nominal	Nominal	Mean Measured		n	%±SD	n	%±SD
Contorol			30	0	0	0	0
DMF Control			30	0	0	0	0
0.18	0.15	0.15	30	0	0	0	0
0.36	0.30	0.24	30	0	0	0	0
0.73	0.60	0.50	30	0	0	0	0
1.46	1.20	1.07	30	0	0	7	23±27
2.91	2.40	2.18	30	5	17±15	26	87±16
5.83	4.80	4.04	30	9	30±11	28	93±10

 

 

 

 

 

 

 

 

 

 

*) DMF (0.1 mL/L dimethylformamide)

 

 

 

 

 

Table 3

 Statistical Results of Probit Analysis Conducted for Determination of EC50 Values:

Probit Analysis for Data obtained after	Slope Function (after Litchfield & Wilcoxon)	EC 50 mg a.s./L mean meas.	lower 95% cl mg a.s./L mean meas.	upper 95% cl mg a.s./L mean meas.
24 hours	2.2	5.66	3.48	n.d.
48 hours	1.6	1.50	1.28	1.76

n.d.: not determined due to mathematical reasons

Validity criteria fulfilled:

yes

Conclusions:

The aim of the study was to determine the influence of propinep on mobility of Daphnia magna over 48 hours in a static exposure, expressed as EC50 for immobilisation.
First instars of Daphnia magna <24 h old were exposed to nominal concentrations of 0, 0.15, 0.30, 0.60, 1.20, 2.40 and 4.80 mg a.s./L. Based on mean-measured concentrations of propineb, the EC50 for immobilisation after 48 hours was determined to be 1.50 mg a.s./L (95% confidence limits 1.28 -1.76 mg a.s./L).

Executive summary:

The aim of the study was to determine the influence of propinep on mobility of Daphnia magna over 48 hours in a static exposure, expressed as EC₅₀ for immobilisation. First instars of Daphnia magna <24 h old were exposed to nominal concentrations of 0, 0.15, 0.30, 0.60, 1.20, 2.40 and 4.80 mg a.s./L. Based on mean-measured concentrations of propineb, the EC₅₀ for immobilisation after 48 hours was determined to be 1.50 mg a.s./L (95% confidence limits 1.28 -1.76 mg a.s./L).

Description of key information

The aim of the study was to determine the influence of propineb on the mobility of Daphnia magna over 48 hours in a static exposure, expressed as EC₅₀ for immobilisation. The main guideline used for this study was OECD 202. Based on mean-measured concentrations of propineb, the 48-h EC₅₀ for was determined to be 1.50 mg a.s./L (95 % confidence limits 1.28 -1.76 mg a.s./L).

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Remarks:

48 hours

Effect concentration:

1.5 mg/L

Additional information

Another study not included in the summary, was performed by Bruns 2010. The study was performed, to detect possible effects of the test item on mobility of Daphnia magna caused by 48 hours of exposure in a static laboratory test system. Daphnia magna (1^st instars <24 h old were exposed to nominal concentrations of 0, 0.63, 1.25, 2.50, 5.00 and 10.0 mg form./L. No immobility or other effects on behaviour occurred in untreated control within 48 hours of exposure. Based on nominal concentrations of propineb formulation the 48-h EC₅₀ was determined to be 2.85 mg a.s./L.