Alcohols, C12-14, ethoxylated, sulfates, sodium salts

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

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))

Version / remarks:

2004

Qualifier:

according to guideline

Guideline:

ISO 11268-2 (Effects of Pollutants on Earthworms. 2. Determination of Effects on Reproduction)

Version / remarks:

1998

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

no

Details on preparation and application of test substrate:

- Method of mixing into soil: test item was weighed in directly on quartz sand, mixed and moistened. Then the test item was mixed into artifical soil.
- Controls: 4 replicates

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

TEST ORGANISM
- Common name: earthworm
- Source: laboratory rearing stock maintained in the testing facility
- Age at test initiation: at least two months old, but not older than 1 year with a clitellum (age of the worms did not idffer by more than four weeks)
- Weight at test initiation (range): between 350 and 450 mg

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

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

no

Total exposure duration:

56 d

Test temperature:

20.1 - 23.9 °C

pH:

study inititation: 6.4 - 6.5
study end: 6.7 - 6.9

Moisture:

water content at study initiation: 24.2 - 25.4 %
water content at study end: 24.1 - 26.6 %

Details on test conditions:

TEST SYSTEM
- Test container (material, size): BELLAPLAST, 17 cm x 12.5 cm x 6 cm
- Amount of soil or substrate: 749 g moistened substrate ( corresponding to 600 g dry substrate)
- 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 (if soil)
- Composition (if artificial substrate):
- 10 % sphagnum peat (close to pH 5.5-6.0; no visible plant remains, finely ground, air dry)
- 20 % kaolin clay (kaolinite content above 30%)
- 70 % industrial sand (fine sand dominant with more than 50% of the particles between 50 and 200 microns)
- Maximum water holding capacity (in % dry weight): 40.36%
- Pretreatment of soil: after mixing the components, i.e. before pre-moistening, 10 g air-dried finely ground cow manure per 1 kg substrate were added
- Storage (condition, duration): in a closed container

OTHER TEST CONDITIONS
- Photoperiod: 16 h light, 8 h darkness
- Light intensity: 500 - 650 lux

EFFECT PARAMETERS MEASURED: Mortality, and body weight change were recorded after 28 days; number of offspring were determined after 56 days

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.8
- Range finding study: yes
- Test concentrations: control, 150, 225, 338, 506 mg a.i./kg soil dry weight
- Results used to determine the conditions for the definitive study: no mortality after 28 days, mean weight change between 14.3 and 31.2%

Nominal and measured concentrations:

nominal test concentrations: 72, 129, 232, 417, 750 mg i.a/kg soil dry weight

Reference substance (positive control):

yes

Remarks:

Carbendazim

Duration:

56 d

Dose descriptor:

NOEC

Effect conc.:

>= 750 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: reproduction and body weight

Duration:

28 d

Dose descriptor:

EC50

Effect conc.:

> 750 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality

Details on results:

- Mortality at end of exposure period: none
- Changes in body weight of live adults (% of initial weight) at end of exposure period: 49.1, 44.4, 43.2, 52.3 and 58.5 % in the test item treatment groups at 72, 129, 232, 417 and 750 mg a.i./kg soil dry weight; in the control the average body weight change was 49.7 %. The body weight change was not statistically significantly different from the control in any of the test item treatment groups.
- Morphological abnormalities: none
- Behavioural abnormalities: none

Results with reference substance (positive control):

In a test conducted from 24 May to 20 Aug 2011 a statistically significant reduction in reproduction of 68% was observed.

Table 1 Effect on mortality, body weight change and reproduction

Test concentration[mg a.i./kgdw]	Contro1	72	129	232	417	750
Mortality [%]	0	0	0	0	0	0
Mean weight change [%]	49.7	49.1	44.4	43.2	52.3	58.5
Juveniles/replicate [mean]	124.4	103.8	97.0	138.5	117.3	111.3
Reproduction[%] ( deviationfrom control)	-	-16.6	-22.0	+11.3	-5.71	-10.5

Table 2: Number of juvenile worms per vessel after 56 days of exposure to the test item

Texapon N 70 [mg a.i./kg sdw]	Number of juvenile worms per replicate (n=10)								Mean value	±SD	CV [%]	Difference to control [%]
	1	2	3	4	5	6	7	8
Control	107	108	110	118	168	124	113	147	124.4	21.9	17.6	-
72	103	118	92	102	-	-	-	-	103.8	10.7	10.3	-16.6
129	85	128	104	71	-	-	-	-	97.0	24.7	25.5	-22.0
232	152	156	144	102	-	-	-	-	138.5	24.8	17.9	+11.3
417	102	105	124	138	-	-	-	-	117.3	16.9	14.4	-5.71
750	105	128	123	89	-	-	-	-	111.3	17.8	16.0	-10.5

SD = Standard Deviation                            sdw = soil dry weight         
CV = Coefficient of Variation

Table 3: Validity criteria for OECD 222 (2016)

Criterion from the guideline to be satisfied in the controls	Outcome	Validity criterion fulfilled
Adult mortality over the initial 4 weeks of the test is ≤ 10%.	0%	yes
Each replicate (containing 10 adults) to have produced ≥ 30 juveniles by the end of the test.	107-168	yes
The coefficient of variation of reproduction is ≤ 30%.	17.6%	yes

Description of key information

NOEC (56 d)≥750 mg a.i./kg soil dw (OECD 222)

Key value for chemical safety assessment

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

750 mg/kg soil dw

Additional information

The toxicity of Alcohols, C12-14, ethoxylated, sulfates, sodium salts to earthworms was investigated in a GLP guideline study conducted according to OECD 222. Eisenia fetida were exposed to nominal test concentrations of 72, 129, 232, 417, and 750 mg i.a/kg soil dry weight in artificial soil. The total study duration was 56 days. After 28 days mortality and body weight change of adult worms was recorded. After 56 days the number of offspring was determined. No mortality and no significant changes in body weight change and reproduction rate were overserved. The determined NOEC (56 d) was≥750 mg a.i./kg soil dw.