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Ecotoxicological information

Toxicity to soil macroorganisms except arthropods

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Endpoint:
toxicity to soil macroorganisms except arthropods: short-term
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
Oct. 4, 1989-Oct. 18, 1989
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 207 (Earthworm, Acute Toxicity Tests)
GLP compliance:
yes (incl. certificate)
Details on sampling:
Samples were from the 250 mg/kg dw concentration taken on day 0, 7, and 14. Measured concentrations were 235, 211, and 40.5 mg/kg dw respectively.
Vehicle:
no
Details on preparation and application of test substrate:
Test material was mixed with 1.16 L of deionized water, and then added to 2.44 kg of artificial soil.
Test organisms (species):
Eisenia fetida
Animal group:
annelids
Details on test organisms:
TEST ORGANISM
- Common name: Earthworm
- Source: Vittor and Associates, Mobile Alabama
- Age at test initiation (mean and range, SD): adult
- Weight at test initiation (mean and range, SD): average 0.66 g

Study type:
laboratory study
Substrate type:
artificial soil
Total exposure duration:
14 d
Test temperature:
19-22 degrees C
pH:
5.5-5.8
Moisture:
34-36%
Details on test conditions:
TEST SYSTEM
- Test container (material, size): jars with perforated lids
- Amount of soil or substrate: 900 g moist soil
- 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)
- Soil texture (if natural soil)
- % sand: 70
- % clay: 20, also 10% sphagnum peat
- Soil classification system: sandy loam


OTHER TEST CONDITIONS
- Photoperiod: constant lighting
- Light intensity: ~600 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality, health, behaviour, and number of cocoons on days 7 and 14

Nominal and measured concentrations:
Nominal: 0, 63, 125, 250, 500, and 1000 mg/kg dw
Duration:
14 d
Dose descriptor:
NOEC
Effect conc.:
250 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: body weight
Key result
Duration:
14 d
Dose descriptor:
LC50
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
14 d
Dose descriptor:
EC10
Effect conc.:
277 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: body weight
Remarks on result:
other: EC10 was not reported in original study report, but later calculated using study results using the method of Vanewijk, PH, and Hoekstra, JA, 1993. Calculation of the EC50 and its confidence interval when subtoxic stimulus is present. Ecotoxicol. Environ
Details on results:
There was 5% mortality at the 1000 mg/kg dw concentration on day 14. Body weight was decreased in the 500 and 1000 mg/kg dw groups by 23 and 33% respectively. The 1000 mg/kg dw group also showed lethargy and burrowing was also affected. Cocoon production was diminished in the 250 mg/kg dw group, and inhibited at higher concentrations.

Mortality and Weight Loss of Earthworms

Concentration

Number of

% Weight Loss

(mg/kg dw)

Day 0

Day 7

Day 14

Day 0-14

Control

40

40

40

14

63

40

40

40

5

125

40

40

40

6

250

40

40

40

5

500

40

40

40

23

1000

40

38

37

33

Conclusions:
The 14-day LC50 to earthworms was > 1000 mg/kg dw soil. The 14-day NOEC was 250 mg/kg dw soil.
Executive summary:

Groups of earthworms were exposed to concentrations of 0, 63, 125, 250, 500, and 1000 mg/kg dw for 14 days. The test animals were examined for mortality, behaviour, health, and reproduction on days 7 and 14. 5% mortality was seen at the highest concentration. Effects to health were seen at the 500 and 1000 mg/kg dw concentrations. The 14 -day LC50 was > 1000 mg/kg dw, and the NOEC was 250 mg/kg dw.

Endpoint:
toxicity to soil macroorganisms except arthropods: long-term
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Reason / purpose:
reference to same study
Qualifier:
equivalent or similar to
Guideline:
other: Draft ISO/WD 16387 protocol
Qualifier:
equivalent or similar to
Guideline:
ISO 11268-2 (Effects of Pollutants on Earthworms. 2. Determination of Effects on Reproduction)
Deviations:
yes
Remarks:
limited cocoons Comparable to ISO 11268-2, but with only weight measurement
Principles of method if other than guideline:
Adult E. albidus (10) were placed in soil containing the test substance for 21 days. The adults were removed, and the cocoons incubated in the soil for another 21 days. The juveniles were then counted.
GLP compliance:
not specified
Analytical monitoring:
not specified
Details on sampling:
- Sample storage conditions before analysis: The soil samples were stored in glowed glass jars at -20°C until analysis.
Vehicle:
not specified
Details on preparation and application of test substrate:
Earthworm tests: For the reproduction tests, 1 kg dry weight of soil was carefully mixed with 160 mL of LAS solution using an electric mixer and filled into plastic pots. For the growth test with juvenile A. caliginosa, 60 g dry weight of soil were mixed with 9.6 mL of LAS solution with a spatula and filled into 160-mL polyethylene beakers with perforated lids for ventilation.
Enchytraeid test: Forty grams dry weight of soil were mixed with 6.4 mL of LAS solution and filled into 160-mL beakers with perforated lids for ventilation.
Test organisms (species):
other: Enchytraeus albidus, Aporrectodea caliginosa, Aporrectodea longa
Animal group:
annelids
Details on test organisms:
TEST ORGANISM
- Common name: Earthworm (Aporrectodea caliginosa and Aporrectodea longa) and potworm (Enchytraeus albidus)
- Source: Earthworms were collected from an agricultural site near Aarhus, Denmark
- Age at test initiation: Adults for reproduction test and (2-3 weeks old) for the growth test with juvenile A. caliginosa
- Food: The earthworms were fed ad libitum on a mixture of dried cow dung and test soil (1:1 [dry vol %]) subsequently moistened to a 50% water content. Potworms were fed boiled and rolled oats that were subsequently dried and ground.

ACCLIMATION
- Acclimation period: 7 days for earthworms
- Acclimation conditions: Yes
Study type:
laboratory study
Substrate type:
natural soil
Remarks:
(Sandy, agricultural soil)
Limit test:
no
Remarks:
Total exposure duration was 21-42 days
Post exposure observation period:
21 days (28 days for A. caliginosa and A. longa growth tests)
Aporrectodea caliginosa and Aporrectodea longa reproduction tests: 21 days
Aporrectodea caliginosa growth test: 28 days
Aporrectodea longa growth test: 42 days
Enchytraeus albidus reproduction test: 42 days
Details on test conditions:
EARTHWORM TESTS

TEST SYSTEM
- Test container (material, size): 1100 mL plastic pots for the reproduction tests and 160 mL polyethylene beakers for growth tests
- Amount of soil or substrate: 1 kg of dry soil for the reproduction tests and 60 g of dry soil for growth tests
- No. of organisms per container (treatment): Three A. longa, four for A. caliginosa for reproduction test and one for growth test
- No. of replicates per treatment group: 4 for reproduction test and 15 for growth test
- No. of replicates per control: 4 for reproduction test and 15 for growth test
- No. of replicates per vehicle control: No vehicle control used

The six treatments consisted of one control and five concentrations of LAS and these treatments were randomly assigned to the experimental units. After 24-hour equilibration of the test soil, 3 (rather than 10) earthworms were added to closed containers with perforated lids for ventilation. Approximately 5 g per worm were added after the test animals had been introduced. The containers were then incubated for 21 days in darkness and the contents were later wet sieved through a 1-mm mesh. Water content was adjusted after 14 days. For the growth test with juvenile, six treatments consisted of one control and five concentrations of LAS and these treatments were randomly assigned to the experimental units. After 24-hour equilibration of the test soil, one earthworm was added to each container. The beakers were incubated for 28 days in darkness and then the earthworms were recovered and their guts were cleared. The surviving animals were dried for 24 hours and their dry weight was recorded to the nearest 0.1 mg. The examination of the effects on growth of A. longa used the same method except the test period was 42 days.

ENCHYTRAEID TEST

TEST SYSTEM
- Test container (material, size): 160 mL beakers
- Amount of soil or substrate: 40 g of dry soil
- No. of organisms per container (treatment): 10

After 24-hour equilibration of the test soil, 10 adult E. albidus were added to each container and incubated in darkness for 21 days. After incubation, the surviving adult animals were removed from the soil. Now only containing cocoons, the soil was incubated in the beakers for another 21 days to allow development and hatching of the juveniles. After this period, the soil containing juveniles was stained with Bengal red, and water was added to facilitate counting of the juveniles.

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Geographic location: The sandy agricultural soil was collected from Askov experimental station, Danish Institute of Agricultural Sciences, Denmark
- Soil texture (if natural soil)
- % sand (fine and coarse): 82.7
- % silt: 8.6
- % clay: 6.2
- % humus: 2.7
- pH: 5.5
- Soil taxonomic classification: Sandy soil
- Cation-exchange capacity: 3.2 mEq/100 g
- Pretreatment and storage of soil: The soil was dried at 80°C, sifted through a 2-mm mesh, and stored at room temperature until use.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
1. Earthworm reproduction tests: Cocoons and earthworms were counted.
2. Earthworm growth tests: Dry weight was determined.
3. Enchytraeid reproduction test: Surviving animals and cocoons were counted. In addition, development and hatching of the juveniles was also evaluated.

VEHICLE CONTROL PERFORMED: no
Nominal and measured concentrations:
Enchytraeid: The test concentrations were not provided but can be estimated from Figure 5 to be 0, 20, 40, 80, 200 and 400 mg/kg with the numbers of adults surviving per replicate to be approximately 10, 10, 10, 10, 9, and 6, and the numbers of juveniles per replicate (reproduction) to be approximately 77, 50, 37, 21, 0, and 0, respectively.
Reference substance (positive control):
not specified
Key result
Duration:
42 d
Dose descriptor:
EC10
Effect conc.:
6 ng/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Remarks on result:
other: Enchytraeus albidus (95% CL: 3-10 mg/kg soil dw)
Key result
Duration:
21 d
Dose descriptor:
LC10
Effect conc.:
194 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks:
(adults)
Remarks on result:
other: Enchytraeus albidus (95% CL: 135-243 mg/kg soil dw)
Key result
Duration:
21 d
Dose descriptor:
LC10
Effect conc.:
329 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks:
(adults)
Remarks on result:
other: Aporrectodea caliginosa and Aporrectodea longa
Key result
Duration:
21 d
Dose descriptor:
EC10
Effect conc.:
14 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Remarks:
(cocoon production)
Remarks on result:
other: Aporrectodea caliginosa (95% CL: 0-150 mg/kg soil dw)
Key result
Duration:
28 d
Dose descriptor:
LC10
Effect conc.:
> 397 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks:
(juveniles)
Remarks on result:
other: Aporrectodea caliginosa
Key result
Duration:
28 d
Dose descriptor:
EC10
Effect conc.:
105 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
(juveniles)
Remarks on result:
other: Aporrectodea caliginosa (95% CL: 0-244 mg/kg soil dw)
Key result
Duration:
21 d
Dose descriptor:
EC10
Effect conc.:
27 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Remarks:
(cocoon production)
Remarks on result:
other: Aporrectodea longa (95% CL: 5-50 mg/kg soil dw)
Key result
Duration:
42 d
Dose descriptor:
LC10
Effect conc.:
296 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks:
(juveniles)
Remarks on result:
other: Aporrectodea longa
Key result
Duration:
42 d
Dose descriptor:
EC10
Effect conc.:
84 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
(juveniles)
Remarks on result:
other: Aporrectodea longa (95% CL: 5-163 mg/kg soil dw)
Details on results:
Earthworm tests: The LAS had no effect on survival of earthworms at concentrations up to approximately 300 mg/kg, but at 793 mg/kg, all animals died. Despite a rather high variation in data, reproduction in both A. caliginosa and A. longa was much more sensitive than survival to LAS. The reproduction was reduced by 50% at approximately 129 mg/kg and 137 mg/kg respectively.

Enchytraeid test:
Survival of adult enchytraeids was not affected at concentrations up to 250 mg/kg, but at 430 mg/kg, a 50% reduction in survival was observed. Also in this species, reproduction was a much more sensitive parameter than survival, showing a 50% reduction at approximately 41 mg/kg.


It should be noted that these studies report worst case exposures due to the use of a sandy test soil and the fact that LAS was added as an aqueous solution to the soil. In addition, too few replicates were used for the ECx approach (e.g., <5 controls) and several key deviations from draft protocols limited the reliability of endpoints for some studies (e.g., A. caliginosa and A. longa cocoon production). Nominal concentrations were derived from tables and figures since actual values were not found in the text.

The following table shows the results of all tests.  All values are nominal LAS concentrations in mg/kg dry weight.

Species

Parameter

NOEC

LOEC

LC10 or EC10

LC50 or EC50

Reliability Rating

Rationale for Reliability Rating

Enchytraeus albidus

Survival, adults

Reproduction

198

20

397

40

194

6

430

41

1

1

Draft ISO/WD 16387 protocol

Aporrectodea caliginosa

Survival, adults

Cocoon production

Survival, juveniles

Growth, juveniles

278

>793

>397

278

793

>793

>397

397

329

14

>397

105

535

129

>397

354

2

3

2

2

Comparable to

ISO 11268-2

Deviations; limited cocoons

Comparable to ISO 11268-2, but with only weight measurement

Aporrectodea longa

Survival, adults

Cocoon production

Survival, juveniles

Growth, juveniles

278

>793

397

79

793

>793

793

278

329

27

296

84

535

137

517

349

2

3

2

2

Comparable to

 ISO 11268-2

Deviations; limited cocoons

Comparable to ISO 11268-2, but with only weight measurement

Folsomia fimetaria

Survival, adults

Reproduction

>793

278

>793

278

>793

85

>793

424

1

1

Comparable to  ISO 11267

Hypogastrura assimilis

Reproduction

79

278

99

421

1

Comparable to  ISO 11267

Hypoaspis aculeifer

Survival, adults

Reproduction

>793

278

>793

793

>793

82

>793

236

2

2

No guideline available

Validity criteria fulfilled:
yes
Conclusions:
In the three reproduction tests of sodium salt of linear alkyl benzene sulphonates (C10-C13) with Enchytraeus albidus, Aporrectodea caliginosa and Aporrectodea longa, Enchytraeus albidus was the most sensitive species with the 42-day EC10 value of 6 mg/kg dry weight, based on reproduction. The 21-day EC10 values for cocoon production for Aporrectodea caliginosa and Aporrectodea longa were 14 and 27 mg/kg dry weight, respectively. The LC10 values were 194, 329 and 329 mg/kg dry weight for Enchytraeus albidus, Aporrectodea caliginosa and Aporrectodea longa, respectively. Reproduction was a much more sensitive parameter than survival in all the three species.

In juvenile growth test with the earthworm species, the EC10 values were 105 and 84 mg/kg dry weight, based on growth and the LC10 values were >397 and 296 mg/kg dry weight, based on survival for Aporrectodea caliginosa and Aporrectodea longa, respectively.
Executive summary:

The aim of study was to study the effects of sodium salt of linear alkyl benzene sulphonates (C10-13 LAS) on the reproduction of three soil invertebrates’ species (earthworms and potworms) and growth of two earthworm species.

For the reproduction tests, adult Enchytraeus albidus, Aporrectodea caliginosa and Aporrectodea longa were exposed to control and five concentrations of LAS for 21, 21 and 42 days, respectively and the offspring’s were counted at the end of study. For the growth test, juvenile Aporrectodea caliginosa and Aporrectodea longa were exposed to control and five concentrations of LAS for 28 and 42 days, respectively. Survival and dry weight were determined at end of exposure.

The LAS had no effect on survival of earthworms at concentrations up to approximately 300 mg/kg, but at 793 mg/kg,all animals died. In both species, the reproduction was reduced by 50% at approximately 130 mg/kg. Growth of juvenile Enchytraeus albidus individuals was reduced by 50% at approximately 350 mg/kg. Survival of adult enchytraeids was not affected at concentrations up to 250 mg/kg, but at 400 mg/kg, a 50% reduction in survival was observed. Also in this species, reproduction was a much more sensitive parameter than survival, showing a 50% reduction at approximately 40 mg/kg.

In the three reproduction tests of sodium salt of linear alkyl benzene sulphonates (C10-C13) with Enchytraeus albidus, Aporrectodea caliginosa and Aporrectodea longa, Enchytraeus albidus was the most sensitive species with the 42-day EC10 value of 6 mg/kg dry weight, based on reproduction. The 21-day EC10 values for cocoon production for Aporrectodea caliginosa and Aporrectodea longa were 14 and 27 mg/kg dry weight, respectively. The LC10 values were 194, 329 and 329 mg/kg dry weight for Enchytraeus albidus, Aporrectodea caliginosa and Aporrectodea longa, respectively. Reproduction was a much more sensitive parameter than survival in all the three species.

In juvenile growth test with the earthworm species, the EC10 values were 105 and 84 mg/kg dry weight, based on growth and the LC10 values were >397 and 296 mg/kg dry weight, based on survival for Aporrectodea caliginosa and Aporrectodea longa, respectively.

Endpoint:
toxicity to soil macroorganisms except arthropods: long-term
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Reason / purpose:
reference to same study
Qualifier:
according to
Guideline:
ISO 11268-2 (Effects of Pollutants on Earthworms. 2. Determination of Effects on Reproduction)
Deviations:
no
Qualifier:
according to
Guideline:
other: ISO 16387:2004 (Soil quality – Effects of pollutants on Enchytraeidae (Enchytraeus sp.) – Determination of effects on reproduction and survival)
Deviations:
no
GLP compliance:
no
Analytical monitoring:
yes
Details on sampling:
Initial concentrations were measured.
Vehicle:
yes
Details on preparation and application of test substrate:
LAS was mixed in demineralized water, and then added to dry Askov soil. Fresh sludge (approx. 200 g) was freeze-dried. This was crushed and sieved through a 2 mm sieve. A methanol solution of LAS was then added to the sludge.
Test organisms (species):
other: Aporrectodea caliginosa, Enchytraeus crypticus
Animal group:
annelids
Details on test organisms:
- Common name: Aporrectodea caliginosa (earthworm) and Enchytraeus crypticus (enchytraeid)
- Source: The earthworms were collected from surrounding of National Environmental Research Institute, Silkeborg, Denmark. Enchytraeid was collected from permanent laboratory cultures and bred on agar.
- Food: Earthworms were fed 5 g mixture of cow dung and test soil (1:1 [dry vol%]) subsequently moistened to 50% water content.

ACCLIMATION
- Acclimation period: The earthworms were acclimated for up to 7 days in Askov soil used for the tests
Study type:
laboratory study
Substrate type:
natural soil
Remarks:
(Askov sandy loam soil)
Limit test:
no
Total exposure duration:
28 d
Test temperature:
16°C
Details on test conditions:
Aporrectodea caliginosa (Oligochaeta)
TEST SYSTEM
- Test container (material, size): Plastic pots and closed with perforated lids allowing ventilation
- Amount of soil or substrate: 1 kg soil and 80 ml test solution
- No. of organisms per container (treatment): 4
- No. of replicates per treatment group: 6
- No. of replicates per control: 2

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)

SOIL
- Geographic location: The soil was a Danish sandy loam collected from Askov, Jutland.
- Soil texture (if natural soil)
- % sand (Fine and coarse): 76
- % silt: 12
- % clay: 11
- Soil classification system: sandy loam
- Organic carbon (%):1.3
- Humus (%): 2.5
- pH: 6.18
- Potassium (meq/100 g): 0.50
- Sodium (meq/100 g): 0.09
- Magnesium (meq/100 g): 0.48
- Calcium (meq/100 g): 5.6
- CEC: 13.4
- Pretreatment of soil: The soil was defaunated by heating to 80°C for 24 h in an oven

SLUDGE
The sludge was collected from the Skaevinge Waste Water Treatment Plant in Denmark receiving
waste water predominantly from households.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Cocoons were collected by sieving with 1 mm mesh after 28 days.

Enchytraeus crypticus (Enchytraeidae)

- Amount of soil or substrate: 20 g soil and 3 ml test solution
- No. of replicates per treatment group: 3

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)

SOIL
- Geographic location: The soil was a Danish sandy loam collected from Askov, Jutland.
- Soil texture (if natural soil)
- % sand: 76
- % silt: 12
- % clay: 11
- Soil classification system: sandy loam
- Organic carbon (%):1.3
- Humus (%): 2.5
- pH: 6.18
- Potassium (meq/100 g): 0.50
- Sodium (meq/100 g): 0.09
- Magnesium (meq/100 g): 0.48
- Calcium (meq/100 g): 5.6
- CEC: 13.4
- Pretreatment of soil: The soil was defaunated by heating to 80°C for 24 h in an oven

SLUDGE
The sludge was collected from the Skaevinge Waste Water Treatment Plant in Denmark receiving
waste water predominantly from households.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Survival of adults and production of juveniles was measured after 28 days.

Nominal and measured concentrations:
LAS DIRECTLY SPIKED TO SOIL TEST:
A. caliginosa: 0 and 25-150 mg/kg soil (nominal concentrations)
E. crypticus 0, 10, 40, 60, 80, 100, 120 mg/kg soil (nominal concentrations)
0.65, 7.1, 26.3, 45.5, 60.3, 92.4, 102.7 mg/kg soil (measured concentrations after incubation for 24 h)

LAS SPIKED TO SLUDGE TEST:
A. caliginosa: 0 and 150 mg a.i./kg soil (nominal concentrations), corresponding to 0 and 92 g a.i./kg freeze-dried sludge
E. crypticus: 0 and 120 mg a.i./kg soil (nominal concentrations) and 2-4 and 90 mg a.i./kg soil (measured concentrations), corresponding to 0 and 72 g a.i./kg freEze-dried sludge
Reference substance (positive control):
no
Key result
Duration:
28 d
Dose descriptor:
EC10
Effect conc.:
46 mg/kg soil dw
Nominal / measured:
nominal
Basis for effect:
reproduction
Remarks on result:
other: A. caliginosa (95% CI: 13-80) mg/kg soil dw
Key result
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
>= 120 mg/kg soil dw
Nominal / measured:
nominal
Basis for effect:
reproduction
Remarks on result:
other: E. crypticus
Details on results:
LAS DIRECTLY SPIKED TO SOIL TEST:

Enchytraeus crypticus: No toxicity (survival of adults and production of juveniles) to E. crypticus was seen at any dose level, including the highest dose level of 120 mg/kg soil.
Aporrectodea caliginosa: Survival of adult A. caliginosa was unaffected by LAS in soil as all adult worms survived in all treatments, whereas a dose-related inhibition of cocoon production was seen in A. caliginosa at the highest concentration of 150 mg/kg soil. The NOEC for reproduction was 40 mg a.i./kg soil. The LOEC was 75 mg a.i./kg soil. The calculated EC10 value was 46 mg a.i./kg soil (reproduction).

LAS SPIKED TO SLUDGE TEST:

- The addition of non-spiked sludge (control sludge) to soil resulted in high stimulation of reproduction for both the tested species.

Enchytraeus crypticus: Significantly reduced reproduction was observed for enchytraeids at 72 g a.i./kg sludge compared to non-spiked sludge. The reproduction by E. crypticus was, however, comparable to the reproduction observed in the control soil without sludge.
A. caliginosa: Sludge spiked with LAS (dose equivalent to 91 g a.i./kg sludge) did not significantly affect the reproduction (fecundity: number of cocoons).

COMPARISON OF SOIL AND SLUDGE SPIKED TESTS:

Compared to LAS directly spiked to soil, the reproductive output of organisms exposed to spiked sludge was not significantly different for Enchytraeus crypticus and was significantly improved for Aporrectodea caliginosa.
Reported statistics and error estimates:
- Linear, quadratic or logistic models were fitted to dose–response data by employing PROC NLMIXED (SAS Institute Inc, 2004) depending on the shape of the curve.
- The EC10 and LC10 values were estimated from initial, nominal concentrations and 95% confidence intervals for the predicted response (significance inferred at a = 5%). Survival was modelled assuming binomially distributed data and reproduction was modelled assuming normally distributed data.
- Determination of NOEC and treatment comparisons were made by ANOVA, furthermore, for Aporrectodea caliginosa the initial fresh weight was included as a covariate.
- Means comparisons of the soil test, including only the control and the maximum LAS concentration, and the sludge test with and without LAS, were compared in a two-way ANOVA with interactions.
- The LAS and substrate effects were analyzed as main factors and simple effects within the soil substrate main effects by making contrasts.
Validity criteria fulfilled:
yes
Remarks:
[ISO 11268-2 (for earthworm) and ISO 16387:2004 (for enchytraeid)]
Conclusions:
In two long term toxicity tests with spiked soil, the 28 day EC10 value of sodium salt of linear alkyl benzene sulphonates (C10-C13) for Aporrectodea caliginosa was 46 mg a.i./kg soil, based on reproduction. The NOEC (reproduction) for Enchytraeus crypticus was ≥120 mg a.i./kg soil. Addition of non-spiked sludge to soil resulted in high stimulation of reproduction for both the species. The spiking of sludge with LAS resulted in reproduction of test species comparable to the control soil without sludge. Therefore, compared to LAS directly spiked to soil, the reproductive output of organisms exposed to spiked sludge was not significantly different for Enchytraeus crypticus and was significantly improved for Aporrectodea caliginosa.
Executive summary:

The toxicity potential of sodium salt of linear alkyl benzene sulphonates (C10-C13) was assessed with two annelids species, Aporrectodea caliginosa (earthworm) and Enchytraeus crypticus (enchytraeid) in a sandy loam soil. The tests were conducted according to ISO 11268-2 (for earthworm) and ISO 16387:2004 (for enchytraeid).

Groups of A. caliginosa were exposed to soil containing 25-150 mg/kg soil of LAS for 28 days. Groups of E. crypticus were exposed to concentrations of 0, 10, 40, 60, 80, 100, or 120 mg/kg soil (nominal). Test concentrations were analytically verified. In addition to LAS directly spiked to soil, tests with LAS spiked to sludge were also performed to study if this would change the toxicity compared with the soil test.

No adverse effect was observed on the survival of adults and production of juveniles for E.crypticus up to the highest dose level of 120 mg/kg soil. Survival of adult A. caliginosa was unaffected by LAS in soil as all adult worms survived in all treatments, whereas a dose-related inhibition of cocoon production was seen in A.caliginosa at the highest concentration of 150 mg/kgsoil.

The addition of non-spiked sludge (control sludge) to soil resulted in high stimulation of reproduction for both the tested species. Significantly reduced reproduction was observed for enchytraeids at 72 g a.i./kg sludge compared to non-spiked sludge, however, it was comparable to the reproduction observed in the control soil without sludge. Sludge spiked with LAS (dose equivalent to 91 g a.i./kg sludge) did not significantly affect the reproduction (fecundity: number of cocoons).

In two long term toxicity tests with spiked soil, the 28 day EC10 value of sodium salt of linear alkyl benzene sulphonates (C10-C13) for Aporrectodea caliginosa was 46 mg a.i./kg soil. The NOEC for Enchytraeus crypticus was ≥120 mg a.i./kg soil. Addition of non-spiked sludge to soil resulted in high stimulation of reproduction for both the species. The spiking of sludge with LAS resulted in reproduction of test species comparable to the control soil without sludge. Therefore, compared to LAS directly spiked to soil, the reproductive output of organisms exposed to spiked sludge was not significantly different for Enchytraeus crypticus and was significantly improved for Aporrectodea caliginosa.

These long term toxicity studies to Aporrectodea caliginosa and Enchytraeus crypticus are classified as acceptable, and satisfies the guideline requirements for the ISO 11268-2 (for earthworm) and ISO 16387:2004 (for enchytraeid).

Endpoint:
toxicity to soil macroorganisms except arthropods, other
Remarks:
(multiple short and long term studies)
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
other: This study was given a reliability score of 4 because the original reports reviewed by the authors were not directly reviewed in the compilation of this robust summary.
Reason / purpose:
reference to same study
Principles of method if other than guideline:
not specified
GLP compliance:
not specified
Analytical monitoring:
not specified
Vehicle:
not specified
Test organisms (species):
other: Eisenia foetida, Lumbricus terrestris, Aporrectodea caliginosa, Aporrectodea longa, Enchytraeus albidus
Animal group:
annelids
Details on test organisms:
eleven soil dwelling invertebrate species
Study type:
other: no data
Substrate type:
not specified
Remarks:
no data
Key result
Dose descriptor:
EC10
Effect conc.:
383 mg/kg soil dw
Basis for effect:
reproduction
Remarks on result:
other: Eisenia foetida
Key result
Dose descriptor:
NOEC
Effect conc.:
667 mg/kg soil dw
Basis for effect:
other: weight
Remarks on result:
other: Lumbricus terrestris
Key result
Dose descriptor:
EC10
Effect conc.:
14 mg/kg soil dw
Basis for effect:
reproduction
Remarks on result:
other: Aporrectodea caliginosa
Key result
Dose descriptor:
EC10
Effect conc.:
27 mg/kg soil dw
Basis for effect:
reproduction
Remarks on result:
other: Aporrectodea longa
Key result
Dose descriptor:
EC10
Effect conc.:
6.2 mg/kg soil dw
Basis for effect:
reproduction
Remarks on result:
other: Enchytraeus albidus
Details on results:
Values were extracted from a variety of original references and compiled for this article.

Table 1: The following table shows the available NOEC, EC10 and EC50values for eleven soil dwelling invertebrate species (in mg/kg dry weight) (Jensen et al., 2001)

                                  

Species

Endpoint

NOEC

EC10

EC50

Eisenia foetida

Reproduction

383

558

Lumbricus terrestris

Weight

667

Aporrectodea caliginosa

Reproduction

14

129

Aporrectodea longa

Reproduction

27

137

Folsomia fimetaria

Reproduction

96

442

Folsomia candida

Reproduction

18

91

Hypoaspis aculeifer

Reproduction

81.7

236

Enchytraeus albidus

Reproduction

6.2

40.5

Platynothrus peltifer

Reproduction

320

467

Isotoma viridis

Growth

41

Hypogastrura assimilis

Reproduction

99.8

421

Conclusions:
The EC10 values of LAS for Eisenia foetida, Aporrectodea caliginosa, Aporrectodea longa, Enchytraeus albidus were 383, 14, 27 and 6.2 mg/kg dry weight. The NOEC for Lumbricus terrestris was 667 mg/kg dry weight.
Executive summary:

Various studies on terrestrial toxicity of LAS were performed with terrestrial organisms. The EC10 values of LAS for Eisenia foetida, Aporrectodea caliginosa, Aporrectodea longa, Enchytraeus albidus were 383, 14, 27 and 6.2 mg/kg dry weight. The NOEC for Lumbricus terrestris was 667 mg/kg dry weight.

Description of key information

A series of studies conducted with LAS on soil macroorganisms are summarized below.

Study 1 (C10-13 LAS):

Groups of earthworms were exposed to C10-13 LAS, sodium salt, concentrations of 0, 63, 125, 250, 500 and 1000 mg/kg soil dw for 14 d. The test animals were examined for mortality, behaviour, health and reproduction on Days 7 and 14. Five percent mortality was seen at the highest concentration. Effects on health were noted at the 500 and 1000 mg/kg soil dw concentrations. The 14 d LC50 was > 1000 mg/kg soil dw and the NOEC was 250 mg/kg soil dw (Swigert, 1989).

Study 2 (C10-13 LAS):

The aim of study was to study the effects of C10-13 LAS, sodium salt (average alkyl chain length C11.6), on the reproduction of three soil invertebrate species (earthworms and potworms) and growth of two earthworm species.

For the reproduction tests, adult Enchytraeus albidus, Aporrectodea caliginosa and Aporrectodea longa were exposed to control and five concentrations of test substance for 21, 21 and 42 d, respectively and the offspring were counted at the end of study.

For the growth test, juvenile Aporrectodea caliginosa and Aporrectodea longa were exposed to control and five concentrations of test substance for 28 and 42 d, respectively. Survival and dry weight were determined at end of exposure.

The test substance had no effect on survival of earthworms at concentrations up to approximately 300 mg/kg soil dw, but at 793 mg/kg soil dw, all animals died. In both species, the reproduction was reduced by 50% at approximately 130 mg/kg soil dw. Growth of juvenile Enchytraeus albidus individuals was reduced by 50% at approximately 350 mg/kg soil dw. Survival of adult enchytraeids was not affected at concentrations up to 250 mg/kg soil dw, but at 400 mg/kg soil dw, a 50% reduction in survival was observed. In this species, reproduction was also a much more sensitive parameter than survival, showing a 50% reduction at approximately 40 mg/kg soil dw.

In conclusion, in the three reproduction tests of C10-13 LAS, sodium salt (average alkyl chain length C11.6), with Enchytraeus albidus, Aporrectodea caliginosa and Aporrectodea longa, Enchytraeus albidus was the most sensitive species with the 42 d EC10 value of 6 mg/kg soil dw, based on reproduction. The 21 d EC10 values for cocoon production for Aporrectodea caliginosa and Aporrectodea longa were 14 and 27 mg/kg soil dw, respectively. The LC10 values were 194, 329 and 329 mg/kg soil dw for Enchytraeus albidus, Aporrectodea caliginosa and Aporrectodea longa, respectively. Reproduction was a much more sensitive parameter than survival in all the three species.

In juvenile growth tests with the earthworm species, the EC10 values were 105 and 84 mg/kg soil dw, based on growth and the LC10 values were >397 and 296 mg/kg soil dw, based on survival for Aporrectodea caliginosa and Aporrectodea longa, respectively (Holmstrup, 2001).

Study 3 (C10-13 LAS):

The toxicity of C10-13 LAS, sodium salt was assessed in two annelids species, Aporrectodea caliginosa (earthworm) and Enchytraeus crypticus (enchytraeid) in a sandy loam soil. The tests were conducted according to ISO Method 11268-2 (for earthworm) and ISO Method 16387:2004 (for enchytraeid). Groups of A. caliginosa were exposed to soil containing 25-150 mg/kg soil dry weight (dw) (nominal) of test substance for 28 d. Groups of E. crypticus were exposed to nominal concentrations of 0, 10, 40, 60, 80, 100 or 120 mg/kg soil dw. Test concentrations were analytically verified. In addition to LAS directly spiked to soil, tests with LAS spiked to sludge were also performed to evaluate if this would change the toxicity compared with the soil test.

Survival of adult A. caliginosa was unaffected by test substance in soil as all adult worms survived in all treatments, whereas a dose-related inhibition of cocoon production was seen in A. caliginosa at the highest concentration of 150 mg/kg soil dw. No adverse effect was observed on the survival of adults and production of juveniles for E. crypticus up to the highest dose level of 120 mg/kg soil dw.

The addition of non-spiked sludge (control sludge) to soil resulted in high stimulation of reproduction for both the tested species. Significantly reduced reproduction was observed for enchytraeids at 72 g a.i./kg sludge compared to non-spiked sludge. However, it was comparable to the reproduction observed in the control soil without sludge. Sludge spiked with test substance (dose equivalent to 91 g a.i./kg sludge) did not significantly affect the reproduction (fecundity: number of cocoons).

In conclusions, in two long term toxicity tests with spiked soil, the 28 d EC10 value of C10-13 LAS, sodium salt, for Aporrectodea caliginosa was 46 mg a.i./kg soil dw. The NOEC for Enchytraeus crypticus was ≥120 mg a.i./kg soil dw. Addition of non-spiked sludge to soil resulted in high stimulation of reproduction for both the species. The spiking of sludge with LAS resulted in reproduction of test species comparable to the control soil without sludge. Therefore, compared to LAS directly spiked to soil, the reproductive output of organisms exposed to spiked sludge was not significantly different for Enchytraeus crypticus and was significantly improved for Aporrectodea caliginosa (Krogh, 2007). 

Study 4 (C10-13 LAS):

Various studies on terrestrial toxicity of C10-13 LAS, sodium salt, were performed with terrestrial organisms. The EC10 values for Eisenia foetida, Aporrectodea caliginosa, Aporrectodea longa and Enchytraeus albidus were 383, 14, 27 and 6.2 mg/kg soil dw. The NOEC for Lumbricus terrestris was 667 mg/kg soil dw (Jensen, 2001).

Key value for chemical safety assessment

Additional information