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Ecotoxicological information

Toxicity to soil macroorganisms except arthropods

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Reference
Endpoint:
toxicity to soil macroorganisms except arthropods: short-term
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1998
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
GLP study according to protocol
Qualifier:
according to
Guideline:
OECD Guideline 207 (Earthworm, Acute Toxicity Tests)
Deviations:
no
Qualifier:
according to
Guideline:
other: CTB guideline section H.4.2
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
no
Vehicle:
no
Details on preparation and application of test substrate:
50, 160, 504, 1606 and 5009 mg of the test substance were each accurately weighed out and dissolved in 1000 mL of "Ultrapur" water, resulting in solutions of 50, 161, 504, 1606 and 5009 m/L, respectively. From each of these solutions 100 mL was added to 675 g of artificial soil to reach concentrations of 10, 32, 100, 320 and 1000 mg of test substance per kg of dry soil.
Test organisms (species):
Eisenia fetida
Details on test organisms:
- Species/strain: Eisenia fetida
- Source: commercial hatchery Blades Biological, Cowden-Edenbridge, Kent, England.
- Culturing techniques: cultured in the laboratory under standard conditions in a horse-manure. garden soil mixture (1 : 1) at about 22- 23°C.
- Pre-treatment: Before use the worms were acclimated for ca. 24 h in the artificial soil. The amount of calcium carbonate was adjusted such that the final pH of the mixture was 6.0 ±0.5. The amount of water added was sufficient to yield a final carbonate content (based on dry constituents) of about 55% after addition of the test substance solutions.
Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
no
Total exposure duration:
14 d
Test temperature:
19.6 - 20.5 °C
pH:
6.1 - 6.3
Moisture:
53.2 - 54.9 %
Details on test conditions:
SOIL
- Source: Various suppliers; 
Sphagnym peat: DEGA, Delft, The Netherlands; 
Industrial sand: Van Loon, Vreeswijk, The Netherlands; 
Kaolin clay: Vingerling bv, Haastrecht, The Netherlands 
- Aeration: no
- ratio Sphagnum peat:kaolin clay: industrial sand: 1:2:7
- pH: 6.0 +/- 0.5 (adjusted using calcium carbonate)
The components were mixed with distilled water and calcium carbonate in a polyethylene bag. The amount of calcium carbonate was adjusted such that the final pH of the mixture was 6.0 ± 0.5. The amount of water added was sufficient to yield a final carbonate content (based on dry constituents) of about 55% after addition of the test substance solutions. pH 6.1 (start and end of the test), moisture 54.9% (start test) 53.2% (end test.)
sphagnum peat consist for 90% of organic matter which means that the artifical soil contained about 5.2% organic carbon.

TEST SYSTEM
- Dosing rate: single
- Exposure vessel type: glass containers of 1000 ml 
- Number of replicates, individuals per replicate: 4 replicates, 10 organisms 
- Light source: Continuous low intensity illumination
- Intensity of irradiation: 577 - 626 lux
- Amount of artificial soil (kg)/ container: 775 g

DURATION OF THE TEST: 14 days  
ENDPOINTS ASSESSED: mortality, abnormalities, body weight
Nominal and measured concentrations:
Nominal: 10-32-100-320-1000 mg/kg dw soil
Reference substance (positive control):
no
Duration:
14 d
Dose descriptor:
NOEC
Effect conc.:
320 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
14 d
Dose descriptor:
LC50
Effect conc.:
503 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: 95% C.L.: 416-608 mg/kg dw
Duration:
14 d
Dose descriptor:
LC100
Effect conc.:
1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
- Effect data: at 1000 mg/kg a 100% mortality was observed. At 320 mg/kg no significant adverse effect was observed. 
- Other effects: Adverse effects with respect to the condition of the test animals were observed at 320 mg/kg dry soil. At that concentration the worms were slower and smaller (visually assessed). The NOEC for condition is therefore 100 mg/kg dry soil.
Weight, was not significantly altered (decreased) at 320 mg/kg dry soil (p = 0.01 ; two-tailed Dunnett-test). The NOEC for weight is therefore 320 mg/kg dry soil. However, weight decrease was significantly different at this concentration. Therefore the NOEC value for weight decrease is given additionally.
Reported statistics and error estimates:
STATISTICS: The NOEC for weight was determined by multiple comparison analyses using two tailed Dunnett's test. A significance level of 1% was employed.

METHOD OF CALCULATION: LC50 was calculated according to the method as described by Kooijman (Kooijman S.A.L.M, Parametric analyses of mortality in bioassays, Water Res. 15, 107-119 (1981)). In addition the LC100 was determined.

Mortality data

Conc.

[mg/kg soil]

Mortality

Number           

   7d                14d

Percentage

       7 d                   14 d

0

0

0

0

0

10

0

0

0

0

32

1

1

2.5

2.5

100

1

1

2.5

2.5

320

0

1

0

2.5

1000

40

40

100

100

Temperature [°C]

19.7

20.5

 

pH

6.2 (0 d)

6.3

 

Moisture content

54.9 (0 d)

53.2

 

Validity criteria fulfilled:
yes
Conclusions:
A dose response related increase in mortality was observed in this study. A very steep increase was seen between 320 and 1000 mg/kg dry soil. Within the control groups there was no mortality and thus the test fulfilled the validity criterion of no more than 10% mortality in the control.
Executive summary:

The acute toxicity of Halamid to the worm Eisenia fetida was determined as described in the OECD Guideline no. 207, the guidelines of the Netherlands Pesticides Bureau and according to the OECD Principles of Good Laboratory Practice.

Worms were exposed to the test substance, mixed with an artificial soil, at concentrations of 10, 32, 100, 320 and 1000 mg/kg of dry soil for a period of 14 days.

The test was carried out with four test containers (fourfold replicates for the control and all concentrations tested). The survival of the test animals was determined and the condition of the worms (appearance and behaviour) was visually assessed after 7 and 14 days of exposure. All worms were individually weighed at the start of the experiment; at the end of the test the weight of the surviving worms was determined for each replicate test container.

The conditions during the experiment were:

Temperature : 19.6 to 20.5 °C

Moisture content at start (based on dry constituents) : 54.9 %

Moisture content at end (based on dry constituents) : 53.2 %

pH at start (control) : 6.1

pH at start (highest test concentration) : 6.2

pH at end (control) : 6.1

pH at end (highest concentration with survival) : 6.3

The summarized results of the test were in mg/kg of dry soil (nominal concentrations; 95% conf. int. between brackets):

7 days LC50 : 527 (435 - 638)

14 days LC50 : 503 (416 -608)

14 days LC100 : 1000

14 days NOEC (survival) : 320

14 days NOEC (appearance and behaviour) : 100

14 days NOEC (weight) : 320

14 days NOEC (weight decrease) : 100

The concentrations quoted in this report refer to the test substance as supplied by the sponsor.

Description of key information

The acute toxicity of Halamid to the worm Eisenia fetida was determined as described in the OECD Guideline no. 207, the guidelines of the Netherlands Pesticides Bureau and according to the OECD Principles of Good Laboratory Practice. Worms were exposed to the test substance, mixed with an artificial soil, at concentrations of 10, 32, 100, 320 and 1000 mg/kg of dry soil for a period of 14 days. The test was carried out with four test containers (four fold replicates for the control and all concentrations tested). The survival of the test animals was determined and the condition of the worms (appearance and behaviour) was visually assessed after 7 and 14 days of exposure. All worms were individually weighed at the start of the experiment, at the end of the test the weight of the surviving worms was determined for each replicate test container.A dose response related increase in mortality was observed in this study. A very steep increase was seen between 320 and 1000 mg/kg dry soil. Within the control groups there was no mortality and thus the test fulfilled the validity criterion of no more than 10% mortality in the control. The 14 day NOEC for weight and survival were 100 and 320 mg/kg of dry soil, respectively. LC50 after 14 days was determined as 503 mg/kg of dry soil.

Key value for chemical safety assessment

Short-term EC50 or LC50 for soil macroorganisms:
100 mg/kg soil dw

Additional information