6'-(dibutylamino)-3'-methyl-2'-(phenylamino)spiro[isobenzofuran-1(3H),9-(9H)-xanthen]-3-one

Toxicity to soil macroorganisms except arthropods

Link to relevant study record(s)

Reference

Endpoint:

toxicity to soil macroorganisms except arthropods: short-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1991-05-08 to 1991-05-22

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 207 (Earthworm, Acute Toxicity Tests)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on preparation and application of test substrate:

- Method of mixing into soil: The test item was applied as dry sand premixes. The dose concentrations were calculated to give the same weight of sand at all dose levels. For each group the premix was thoroughly mixed with a small proportion of soil prior to incorporation with the main soil bulk. Water was gradually mixed with the treated soil to give a moisture content equivalent to 35 % of the dry weight.
- Controls: Negative control soil was treated with an equivalent weight of dry sand.

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

TEST ORGANISM
- Common name: earthworm
- Source: Local Supplier, Houghton, St. Ives, Cambridgeshire, England, UK
- Age at test initiation: adult with clitella
- Weight at test initiation: 529 - 643 mg

ACCLIMATION
- Acclimation period: 1 day
- Acclimation conditions: same as test

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

no

Total exposure duration:

14 d

Test temperature:

20 - 21 °C

pH:

5.7 (addition of calcium carbonate)

Moisture:

35 % of the dry weight

Details on test conditions:

TEST SYSTEM
- Test container (material, size): glass container (1 L)
- Amount of soil or substrate: 729 g
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 4
- No. of replicates per control: 4

SOURCE AND PROPERTIES OF SUBSTRATE
- Composition (% w/w):
Quartz sand 70
Kaolin clay 20
Sphagnum peat 10

OTHER TEST CONDITIONS
- Photoperiod: continous
- Light intensity: 400 lux

EFFECT PARAMETERS MEASURED:
Counts of worms, on day 7 and 14
Health and mortalities: Daily
Weights: on day 0 and 14

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.8
- Test concentrations: 0, 95, 171, 309, 556, 1000 ppm

Nominal and measured concentrations:

0, 95, 171, 309, 556, or 1000 ppm

Reference substance (positive control):

yes

Remarks:

chloroacetamide

Key result

Duration:

14 d

Dose descriptor:

LC50

Effect conc.:

> 1 000 other: ppm

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

- Mortality at end of exposure period:
No mortalities were detected at the day 7 count. By day 14, one mortality had occurred in the 309 ppm dose group. This mortality was not considered to be treatment-related. No mortalities occurred in any other group.
- Changes in body weight of live adults at end of exposure period: Weight losses were observed in all groups. However, there was no clear evidence of any treatment-related effect.

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- Relevant effect levels: The LC50 values for chloroacetamide, tested previously as a positive control between 15 and 29 May 1985 were found to be:
Day 7 LC50: 43.1 ppm (95 % confidence limits 34.0 – 57.6 ppm)
Day 14 LC50: 24.6 ppm (95 % confidence limits 20.2 – 30.5 ppm)

Reported statistics and error estimates:

As outlined in OECD Method No. 207

Table 1: Weight Development

Treatment	Concentration (ppm)	Weight (mg)				% Decrease
		Day 0	Group mean	Day 14	Group mean
Neg. Control	0	643	592	478	451	23.8
		529		428
		596		442
		600		456
Test Item	95	639	602	478	489	23.8
		601		460
		572		439
		595		459
Test Item	171	605	602	470	479	20.4
		582		432
		628		488
		594		525
Test Item	309	582	603	447	488	19.1
		594		528
		623		496
		613		485
Test Item	556	584	603	492	498	17.4
		594		497
		614		508
		620		496
Test Item	1000	615	601	486	474	21.1
		615		490
		587		449
		585		471

Validity criteria fulfilled:

yes

Conclusions:

It was not possible to determine the LC50 values of ODB-2 to the earthworm at Days 7 and 14. The LC50 is above 1000 ppm, the maximum treatment level tested.

Executive summary:

In a 14 -day acute toxicity study, earthworms (Eisenia fetida) were expsoed to the test substance at 0, 95, 171, 309, 556, or 1000 ppm (Huntingdon, 1991). The reference chemical used was chloroacetamide. The experiment was carried out in accordance with OECD guideline 207.

No mortalities were detected at the day 7 count. By Day 14, one mortality had occurred in the 309 ppm dose group. This mortality was not considered to be treatment-related. No mortalities occurred in any other group. Weight losses were observed in all groups. However, there was no clear evidence of any treatment-related effect.

The LC50 values for chloroacetamide, tested previously as a positive control were found to be:

Day 7 LC50: 43.1 ppm (95 % confidence limits 34.0 – 57.6 ppm)

Day 14 LC50: 24.6 ppm (95 % confidence limits 20.2 – 30.5 ppm)

Because of the limited number of mortalities it was not possible to determine the LC50 values of the test item to the earthworm at Days 7 and 14. These values must lie in excess of 1000 ppm, the maximum treatment level used. The test substance is therefore regarded as non-toxic to earthworms above a concentration of 1000 ppm.

Description of key information

It was not possible to determine the LC50 values of the test substance to the earthworm on days 7 and 14. The LC50 is above 1000 ppm, the maximum treatment level tested.

Additional information

In a 14 -day acute toxicity study, earthworms (Eisenia fetida) were exposed to the test substance at 0, 95, 171, 309, 556, or 1000 ppm (Huntingdon, 1991). The reference chemical used was chloroacetamide. The experiment was carried out in accordance with OECD guideline 207.

No mortalities were detected at the day 7 count. By day 14, one mortality had occurred in the 309 ppm dose group. This mortality was not considered to be treatment-related. No mortalities occurred in any other group. Weight losses were observed in all groups. However, there was no clear evidence of any treatment-related effect.

The LC50 values for chloroacetamide, tested previously as a positive control were found to be:

Day 7 LC50: 43.1 ppm (95 % confidence limits 34.0 – 57.6 ppm)

Day 14 LC50: 24.6 ppm (95 % confidence limits 20.2 – 30.5 ppm)

Because of the limited number of mortalities it was not possible to determine the LC50 values of the test substance to the earthworm on days 7 and 14. These values must be above 1000 ppm, the maximum treatment level used.