Propanol, iminobis-, N-(C16-18 and C18-unsatd. alkyl) derivs.

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:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

2009-07-17 to 2009-09-15

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

See section 13.2 for the read-across justification.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

DIN ISO 11268-2: 1998 (E), BBA Guideline (1994)

Deviations:

yes

Remarks:

Earthworms were also fed weekly after removing adult earthworms at day 28. The soil moisture of all test vessels deviated by more than 10 % of the initial value at the end of the test.

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on preparation and application of test substrate:

- Method of mixing into soil (if used): The test item was weighed out and was evenly distributed in 10 g quartz sand per replicate. The treated quartz sand was given to the natural soil. Demineralised water was added to adjust the humidity of the natural soil to a moisture content of 54 % of the maximum water holding capacity.
Thereafter, the test medium was mixed for at least three minutes and about 600 g soil dry weight were filled into each test vessel.

- Controls: Natural soil moistened with demineralised water without test or reference item was used as control medium.

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

TEST ORGANISM
- Common name: Eisenia fetida SAVIGNY 1826, subspecies Eisenia fetida fetida
- Source: Own breeding
- Age at test initiation (mean and range, SD): synchronized earthworm culture, not deviating more than 1 month
- Weight at test initiation (mean and range, SD): 0.46


ACCLIMATION
- Acclimation period: 2 days prior to test start
- Acclimation conditions (same as test or not): same as test
- Health during acclimation (any mortality observed): healthy

Study type:

laboratory study

Substrate type:

natural soil

Limit test:

no

Total exposure duration:

56 d

Remarks:

Removement of adults after 28 days

Test temperature:

20 +/- 2°C

pH:

Application rate
[mg test item/kg soil DW] pH-value
day 0 day 56
Control 6.34 6.43
62.5 6.30 6.46
125 6.32 6.47
250 6.31 6.41
500 6.34 6.39
1000 6.49 6.32

Moisture:

Application rate
[mg test item/kg soil DW] Moisture [%] of dry weight

day 0 day 60
Control 23.4 31.2*
62.5 23.2 31.2*
125 23.0 29.8*
250 23.3 31.0*
500 23.0 29.5*
1000 23.8 33.8*

Details on test conditions:

TEST SYSTEM
- Test container (material, size): Round plastic boxes with an inner diameter of 15.0 cm corresponding to a bottom surface area of 177 cm2 and a height of 14 cm contained about 600 g soil dry weight. Transparent perforated lids enabled sufficient gas exchange, light input and prevented the substrate from drying.
- Amount of soil or substrate: 600 g
- No. of organisms per container (treatment): 10 (40)
- No. of replicates per treatment group: 4
- No. of replicates per control: 8
- No. of replicates per vehicle control: -


SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Geographic location: Speyer, South Germany
- Pesticide use history at the collection site: Pesticides:
Sampling year and 4 former years: None

Plants:
Sampling year (2009): Meadow
2008: Meadow
2007: Meadow
2006: Meadow
2005: Meadow

Fertilization:
No organic fertilization
Sampling year (2009): None
2008: None
2007: 600 kg/ha CaO
+ 250 kg/ha MgO
2006: None
2005: None

- Soil texture (if natural soil)
-sand: 0.63 – 2.0 mm % 0.7 ± 0.2
0.2 – 0.63 mm % 44.5 ± 2.6
0.063 – 0.2 mm % 35.5 ± 2.2
- % silt: 0.02 – 0.063 mm % 5.8 ± 1.1
0.006 – 0.02 mm % 4.0 ± 0.6
0.002 – 0.006 mm % 3.0 ± 0.6
- % clay: < 0.002 mm % 6.4 ± 0.9
- Soil taxonomic classification: loamy sand (lS)
- Soil classification system: German DIN classification
- Organic carbon (%): 2.09 +/- 0.4
- Maximum water holding capacity (in % dry weigth): 46.5 +/- 6.0



OTHER TEST CONDITIONS
- Photoperiod: 16 h
- Light intensity: 400 - 800 lx


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
The bodyweight of the adult earthworms were individually determined at day 0 and day 28.
After the first four weeks the adult earthworms were removed and mortality and morphological changes of the adult worms were recorded. After further four weeks, the number of offspring hatched from the cocoons were counted.
Observation parameter: a) no obvious pathological symptoms
b) no reaction to touching
c) no negative phototactical reaction
d) spasmodic winding
e) spontaneous segmentation and separation
f) yellow excretion from the oral aperture
g) ulcer and skin bleedings
t) dead earthworms



VEHICLE CONTROL PERFORMED: no


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Range finding study: No

Nominal and measured concentrations:

Nominal application rates: 62.5 - 125 - 250 - 500 - 1000 mg test substance/kg soil dry weight (DW)

Reference substance (positive control):

yes

Remarks:

1.0 - 2.5 - 5.0 mg Carbendazim/ kg soil DW

Duration:

56 d

Dose descriptor:

NOEC

Effect conc.:

500 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Remarks:

reduction of body weight

Details on results:

- Mortality at end of exposure period: No mortality was observed in any of the tested test item and control groups
- Total mass of adults at beginning of test: 0.46
- Changes in body weigth of live adults (% of initial weight) at end of exposure period: please refer to the respective table
- No. of offspring produced: please refer to the respective table
- No. of unhatched cocoons: Not determined
- Morphological abnormalities: None
- Behavioural abnormalities: None
- Other biological observations: None

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- Relevant effect levels: 1.0 mg Carbendazim/kg soil DW

Reported statistics and error estimates:

One Way Analysis of Variance (ANOVA) was carried out for the determination of statistically significant differences compared to the control. A Normality Test and an Equal Variance Test were conducted prior to running the ANOVA. P-values for both Normality and Equal Variance Test are 0.05. The a-value for ANOVA test (acceptable probability of incorrectly concluding that there is a difference) is a = 0.05.

Validity criteria fulfilled:

yes

Conclusions:

Under the conditions of this study, Bis (2-hydroxyethyl) oleyl amine (CAS No 25307-17-9) did not induce mortality or pathological symptoms of adult earthworms after an exposure to the test item concentrations of 62.5, 125, 250, 500 and 1000 mg/kg soil dry weight for 28 days. The biomass increase of the adult earthworms after 28 days did not statistically significant differ compared to the control at 62.5, 125, 250 and 500 mg/kg soil dry weight. However, at 1000 mg/kg soil dry weight the biomass increase was statistically significantly reduced compared to the control. In addition, after eight weeks of exposure the earthworm reproduction was not statistically significantly reduced at 62.5, 125, 250 and 500 mg/kg soil dry weight. At 1000 mg/kg soil dry weight the earthworm reproduction was statistically significantly reduced compared to the control.

Thus, the No Observed Effect Concentration (NOEC) of Bis (2-hydroxyethyl) oleyl amine (CAS No 25307-17-9) with respect to mortality, body weight and reproduction of Eisenia fetida fetida after 8 weeks of exposure in artificial soil was determined to be 500 mg/kg soil dry weight.

Executive summary:

Effects of Bis (2-hydroxyethyl) oleyl amine (CAS No 25307-17-9)(Batch no. S001018) on mortality, biomass and the reproductive potential of the earthworm species Eisenia fetida(Savigny) were determined according to the guidelines DIN ISO 11268-2 (1998), OECD 222 (2004) and BBA (1994) from July 17^th to September 15^th, 2009 at Dr.U.Noack-Laboratorienin 31157 Sarstedt, Germany. The study was conducted under static conditions over 8 weeks with five different application rates (62.5 – 125 – 250 – 500 – 1000 mg/kg soil dry weight), which were mixed into natural soil. Eighty sexually mature earthworms were inserted into 8 control replicates (10 earthworms per replicate) and forty sexually mature earthworms were divided into 4 replicates for the test item concentrations. They had an individual body weight between 0.30 and 0.60 g.

After 28 days no mortality was seen in the control and at all tested application rates (Table 1). Also, no evident pathological symptoms or changes in the behaviour of adult earthworms were seen in the control and at all tested application rates. The earthworm biomass increased in the control and at all tested application rates over 28 days without any statistically significant differences at 62.5, 125, 250 and 500 mg/kg soil dry weight compared to the control. However, at 1000 mg/kg soil dry weight the biomass increase was statistically significant reduced compared to the control. After further four weeks the reproduction rate (average number of juveniles) was 332 in the control and ranged between 254 and 359 at the test item application rates. Compared to the control, the reproduction rate was statistically significantly reduced at 1000 mg/kg soil dry weight. All other tested application rates showed no statistically significant reduction compared to the control (Table 1).

Overall, the NOEC of Bis (2-hydroxyethyl) oleyl amine (CAS No 25307-17-9) concerning mortality, biomass and reproduction of earthworms was determined to be at 500 mg/kg soil dry weight (Table 2).

All validity criteria recommended by the test guidelines were fulfilled.

Summary of All Observed Effects in the Tested Doses

Effects	Bis (2-hydroxyethyl) oleyl amine (CAS No 25307-17-9)
mg/kg soil dry weight	62.5	125	250	500	1000
Mortality of adult earthworms	No	No	No	No	No
Reduction of body weight	No	No	No	No	Yes
Reduced reproduction rate   (Number of juveniles)	No	No	No	No	Yes
Summary of all effects	No	No	No	No	Yes

NOEC and LOEC for Reproduction, Mortality and Body Weight

Bis (2-hydroxyethyl) oleyl amine (CAS No 25307-17-9)
	Application rate
	mg/kg soil dry weight
NOECreproduction, mortality, body weight	500
LOECreproduction, mortality, body weight	1000

Description of key information

No terrestrial test have been performed with di (2-hydroxypropyl)tallow amine. The data available for di (2-hydroxyethyl) oleyl amine (CAS No 25307-17-9) will as a worst-case be used for read-across. Justification for this read-across is presented in IUCLID chapter 13. For Di (2-hydroxyethyl) oleyl amine (CAS No 25307-17-9) one chronic earthworm test has been performed resulting in a NOEC of 500 mg/kg dw.  

Key value for chemical safety assessment

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

500 mg/kg soil dw

Additional information

No effects observed in a chronic terrestrial test up to 500 mg/kg dw.