Quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides

Toxicity to soil macroorganisms except arthropods

Link to relevant study record(s)

Reference

Endpoint:

toxicity to soil macroorganisms except arthropods: long-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From April 20, 2010 to Junne 28, 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 222 (Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei))

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

no

Details on sampling:

Not applicable

Vehicle:

yes

Details on preparation and application of test substrate:

No data

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

TEST ORGANISM
- Common name: Earthworms
- Source: Bred under standardised conditions (see OECD 207, in a breeding medium of cattle manure, peat, sand, calcium carbonate and straw, fed with cattle manure, stored at room temperature) by IBACON
- Age at test initiation (mean and range, SD): 9 to 10 m, with well developed clitellum, age range between test individuals not differing by more than 4 wk
- Weight at test initiation (mean and range, SD): 300- 599 mg

ACCLIMATION
- Acclimation period: 1 d
- Acclimation conditions (same as test or not): In artificial soil, under test conditions
- Health during acclimation (any mortality observed): No data

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

no

Total exposure duration:

8 wk

Remarks:

Exposure Time of Offspring: Additional 28 d

Post exposure observation period:

None

Test temperature:

20- 22 °C

pH:

6.0 -6.2

Moisture:

30.5 - 36.9 % (52.6% -63.6 % of the maximum water holding capacity)

Details on test conditions:

TEST SYSTEM
- Test container (material, size): Plastic boxes (18.3 cm x 13.6 cm x 6 cm, tapered towards the bottom, with a soil surface of approximately 16.5 cm x 11.5 cm = 189.75 cm2 ) with perforated transparent lids to enable exchange of air, to minimise evaporation from the artificial soil, and to prevent the worms from escaping.
- Amount of soil or substrate: 664.6 g of the prepared soil (500 g dry weight plus 159.6 g water plus 5 g food).
- No. of organisms per container (treatment): 10 per replicate, i.e. 80 individuals per control, 40 individuals per test item treated group
- No. of replicates per treatment group: 4
- No. of replicates per control: 8


SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Geographic location: Artificial soil according to OECD 222
- Composition (if artificial substrate): 10% Sphagnum-peat, air-dried and finely ground (2 mm with no visible plant remains); (Floragard, Vertriebs GmbH fur Gartenbau, 26138 Oldenburg, Germany), 20% Kaolin clay (Kaolinite content >30%) (ErbslOh, 65558 Lohrheim, Germany), approximately 0.5% calcium carbonate (CaC03) added to adjust pH to 6.0 ± 0.5 (Merck, 64293 Darmstadt, Germany), approximately 69.5% fine quartz-sand (F34) containing more than 50% by mass of particle size 0.05 mm to 0.2 mm; (Quarzwerke Frechen, Postfach 1780, 50207 Frechen, Germany)
- Organic carbon (%):
- Maximum water holding capacity (in % dry weigth): 58 %

OTHER TEST CONDITIONS
- Photoperiod: 16 h light: 8 h dark
- Light intensity: 400 -800 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality, weight change, feeding activity and reproduction rate were
determined

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2

Nominal and measured concentrations:

Nominal: 0, 62.5, 125, 250, 500, 1000 mg/kg

Reference substance (positive control):

yes

Remarks:

Luxan Carbendazim 500 FC

Key result

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

ca. 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: mortality and weight

Key result

Duration:

56 d

Dose descriptor:

NOEC

Effect conc.:

ca. 500 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Details on results:

- Mortality at end of exposure period: No
Please refer Table 1. in attached background material

Results with reference substance (positive control):

- Results with reference substance valid: Yes
- Relevant effect levels: EC50 for reproduction was calculated as 1.47 mg carbendazim/kg soil dry weight

Reported statistics and error estimates:

Standard procedures, Fisher's Exact Test (mortality), Dunnett's t-test (weight
changes and reproduction)

For result tables, kindly refer to the attached background material section of the IUCLID.

Validity criteria fulfilled:

yes

Conclusions:

Under the study conditions, the NOEC of the test substance for mortality, growth and feeding activity of the earthworm Eisenia fetida was determined to be 1000 mg/kg soil dw (highest tested conc), whereas the NOEC for reproduction was determined to be 500 mg/kg soil dw.

Executive summary:

A study was conducted to evaluate the chronic toxicity of the test substance, C12-18 DAQ (99.6% active) to adult Eisenia fetida, according to OECD guideline 222 and ISO 11268-2 in compliance with GLP. Adult earthworm (Eisenia fetida), with clitellum and weight range 300 to 599 mg and 9 to 10 m old from in-house culture was used in the study. Ten adult earthworm per replicate were allocated to an artificial soil containing 62.5, 125, 250, 500 and 1000 mg a.i./kg soil dw (nominal concentrations) for 56 days. No analytical dose verification was performed. Assessment of adult worm mortality, behavioural effects and biomass development was carried out after 28 d exposure of adult worms in treated artificial soil. Reproduction rate (number of offspring) was assessed after additional 28 d (assessed 56 d after application). No mortality was observed in any treatment group, except at the control, where one worm died. The body weight change of the earthworms after 4 weeks exposure to test material was not statistically significantly different compared to the control up to and including the highest test concentration of 1000 mg test material/kg soil dw. The reproduction rates were not significantly different compared to the control up to and including the test concentration of 500 mg test material/kg dry soil. At the highest test concentration of 1000 mg/kg soil dw the reproduction was significantly reduced compared to the control. No behavioural abnormalities were observed in any of the treatment groups. The feeding activity in all the treated groups was comparable to the control. Under the study conditions, the NOEC of the test substance for mortality, growth and feeding activity of the earthworm Eisenia fetida was determined to be 1000 mg/kg soil dw (highest tested conc), whereas the NOEC for reproduction was determined to be 500 mg/kg soil dw (Witte, 2010).

Description of key information

Based on the results of the study with adult earthworms, the 56 day NOEC of the test substance for effects on reproduction was determined to be 500 mg/kg soil dw (nominal).

Key value for chemical safety assessment

Short-term EC50 or LC50 for soil macroorganisms:

1 000 mg/kg soil dw

Long-term EC10, LC10 or NOEC for soil macroorganisms:

500 mg/kg soil dw

Additional information

A study was conducted to evaluate the chronic toxicity of the test substance, C12-18 DAQ (99.6% active) to adult Eisenia fetida, according to OECD guideline 222 and ISO 11268-2 in compliance with GLP. Adult earthworm (Eisenia fetida), with clitellum and weight range 300 to 599 mg and 9 to 10 m old from in-house culture was used in the study. Ten adult earthworm per replicate were allocated to an artificial soil containing 62.5, 125, 250, 500 and 1000 mg a.i./kg soil dw (nominal concentrations) for 56 days. No analytical dose verification was performed. Assessment of adult worm mortality, behavioural effects and biomass development was carried out after 28 d exposure of adult worms in treated artificial soil. Reproduction rate (number of offspring) was assessed after additional 28 d (assessed 56 d after application). No mortality was observed in any treatment group, except at the control, where one worm died. The body weight change of the earthworms after 4 weeks exposure to test material was not statistically significantly different compared to the control up to and including the highest test concentration of 1000 mg test material/kg soil dw. The reproduction rates were not significantly different compared to the control up to and including the test concentration of 500 mg test material/kg dry soil. At the highest test concentration of 1000 mg/kg soil dw the reproduction was significantly reduced compared to the control. No behavioural abnormalities were observed in any of the treatment groups. The feeding activity in all the treated groups was comparable to the control. Under the study conditions, the NOEC of the test substance for mortality, growth and feeding activity of the earthworm Eisenia fetida was determined to be 1000 mg/kg soil dw (highest tested conc), whereas the NOEC for reproduction was determined to be 500 mg/kg soil dw (Witte, 2010).

The above study is supported by the results from a study with the read across substance C16 -18 DAQ, which has been reported in the EU RAR on DODMAC (or C18 DAQ) (EU, 2002) and has been summarised below:

In the study, Eisenia fetida was exposed to C16-18 DAQ (solved in methanol, 76.1% active ingredient) incorporated into artificial soil consisting of 9.4% organic matter, 70% fine sand and 20% kaolinite clay (Coulson et al. (1989). At the only concentration of the definitive test with 1,000 mg active ingredient/kg dry soil no mortality, no significant reduction in body weight nor any behavioral effects were observed after 14 days.