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Toxicity to soil macroorganisms except arthropods

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Endpoint:
toxicity to soil macroorganisms except arthropods
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
disregarded due to major methodological deficiencies
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Acceptable, well documented publication which meets basic scientific principles.
Principles of method if other than guideline:
Range finding test to determine toxic effects of aluminium on Eisenia fetida in artificial soil. Responses of earthworms to aluminium at three different pHs were followed in terms of growth, cocoon production and cocoon viability.
GLP compliance:
no
Analytical monitoring:
yes
Vehicle:
no
Details on preparation and application of test substrate:
Artificial soil was made up in acordance with OECD guideline 2007 (1984). pH adjusted using calcium carbonate.
Test organisms (species):
Eisenia fetida
Animal group:
annelids
Details on test organisms:
TEST ORGANISM
- Common name: earthworm
- Source: from a bed at Teagasc, Oak Park, Carlow, belonging to vermi-composting group at Carlow Regional Technical College
- Age at test initiation (mean and range, SD): young, non-clitellate
- Weight at test initiation (mean and range, SD): 100 - 200 mg
- feeding during test: 5 g dry weight of urine free "organic" cow dung


ACCLIMATION
- Acclimation period: 12 weeks
- Acclimation conditions (same as test or not): maintained in horse dung in incubators at 20 °C until the onset of sexual maturity
Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
no
Test temperature:
20 ± 2 °C
pH:
4.0, 5.5, and 6.5 [as pH(CaCl2)]
Moisture:
35 % w/w
Details on test conditions:
TEST SYSTEM
- Test container (material, size): translucent plastic container (l , w, h: 150 x 95 x 95 mm)
- Amount of soil or substrate: 50 mm depth
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 4
- No. of replicates per control: 4


OTHER TEST CONDITIONS
- Photoperiod: 24 h
- Light intensity: 300 Lux
Nominal and measured concentrations:
0, 10, 100, 1000, and 2000 mg Al/kg soil
Reference substance (positive control):
no
Reported statistics and error estimates:
Data analysed using ANOVA and Duncan's Multiple Range Test. Percentages and proportions transformed using the arc sine square root transformation

Controls in the optimal pH range 6.5 - 7.0 produced a mean of 1.48 cocoons per worm per week. At low concentrations of aluminum, i.e. 0, 10, and 100 mg/kg, cocoon production was significantly larger at higher pH values. This relationship was not observed at higher levels of aluminum. At low pH, the numbers of cocoons produced were significantly reduced at higher levels of aluminum. The largest number of cocoons produced per worm per week was in the 100 mg Al/kg treatment at pH 6.8 where 1.63 cocoons were produced. Complete inhibition of cocoon production occurred at 4000 mg/kg at pH 4.2

Thus, irrespective of pH, the lowest number of cocoons were prodcued at highest aluminum concentrations and the largest numbers at 10 - 100 mg Al/kg. Based on the observed reductions in the rate of cocoon production calculation of EC50 values was not possible.

In summary the following conclusions might be drawn: (1) lethal dose at pH 4.2 is between 2000 - 4000 mg Al/kg soil, (2) high levels of Al inhibit cocoon production between pH 4 and 7, (3) intermediate Al levels stimulate cocoon production at pH 6 to 7, (4) low levels of Al inhibit hatching and juvenile production at low pH and (5) cocoon viability and juvenile production are stimulated where parents were kept at intermediate levels of Al at pH 6 to 7.

Endpoint:
toxicity to soil macroorganisms except arthropods: long-term
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
disregarded due to major methodological deficiencies
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Acceptable, well documented publication which meets basic scientific principles.
Principles of method if other than guideline:
Impact of the combination of increased aluminum concentrations and soil acidity was studied in the laboratory using Dendrodrilus rubidus. Survival of the parent generation, its activity, production of cocoons and loss of weight were followed for four months in different treated soils. Hatching success, survival and growth were determined in the filial generation
GLP compliance:
no
Analytical monitoring:
no
Details on preparation and application of test substrate:
Soil prepared by mixing mull (brown forest soil, 15 km E of Lund, horizon A1, pH(KCl) 5.7), sand (pine plantation, horizon C, pH(KCl) 4.0), and cattle dung. The components were preperly treated or collected in order to exclude any unwanted soil fauna and to reduce pathogens. The three components were seperately sieved and then mixed in the ratio 20:70:10 vol. %. After mixing soil pH(KCl) was 4.9 and organic content 3 wt%.
Test organisms (species):
other: Dendrodrilus rubidus
Animal group:
annelids
Details on test organisms:
TEST ORGANISM
- Common name: earthworm
- Source: collected under cow droppings in pine plantation in July
- Age at test initiation (mean and range, SD): mature
- Weight at test initiation (mean and range, SD): 232 mg per worm (mean weight)
Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
no
Total exposure duration:
26 wk
Remarks:
Exposure duration for p-generation was 16 weeks, for the f-generation 48 weeks
Test temperature:
14 °C
pH:
4.2 - 4.9
Moisture:
17 % (dry weight) soil moisture
Details on test conditions:
TEST SYSTEM
- Test container (material, size): cardboard boxes (70 x 70 x 200 mm, 1.0 L)
- Amount of soil or substrate: 500 g
- No. of organisms per container (treatment): 5 (test with p-generation)
- No. of replicates per treatment group: 6
- No. of replicates per control: 6
Nominal and measured concentrations:
10, 25, 50, 100, and 200 mg Al/kg soil (dry weight)
Reference substance (positive control):
no
Reported statistics and error estimates:
Mann-Whitney U-test used to examine survival of adults and juveniles, reproduction and number of hatchlings cocoon. Kruskal-Wallis test used to check differences in prevalence of infested cocoons betwen treated soils. Adult weight loss, activity, hatchability of cocoons, and juvenile growth tested with ANOVA. Kendell's t-test used to estimate correlations between treatment and activity with respect to number of deposited cocoons.

The presence of aluminium in soil did not affect the parameters studied in the parent generation. After 4 months survival ranged between 63 % (pH 4.2) and 90 % (pH 4.6). At pH 4.5 and 4.5 more specimens than in the control remained alive. However, there was no significant difference between controls and Al-series and high Al-concentrations did not result in lower survival than in the controls. Earthworms maintained in soils of high Al-concentrations lost weight in the first month and weighed less than the controls. However, after 2 to 4 months there were no differences in comparison to controls.

For the filial generation responses were observed: Reproduction was significantly decreased in the two highest aluminum concentrations and in the pH series at the two lowest pH levels. Hatchability at 200 mg Al/kg soil (dw) significantly decreased. After 26 weeks of incubation all juveniles produced in the first month and maintained in soils of the two highest Al-concentrations had died. Only a few juveniles hatched from cocoons produced in the second month in soils of 200 mg Al/kg soil (dw). All juveniles hatched in soils of 100 mg Al/kg soil (dw) were dead at the end of experiment.

However, from the parallel pH series it was seen that these effects also occurr, sometimes with stronger effects, with lowered pH values alone.

In summary, the study shows that lowered pH in itself affects D. rubidus and the presence of aluminium does not increase the strength of the response. Lowered pH alone is followed by decreased activity, slightly enhanced weight loss of ageing adult individuals, lowered individual production of cocoons, lowered hatching success of cocoons produced late during the reproductive period, and hampered juvenile growth. Some of these effects might be related, e.g. lowered activity might affect feeding activity and fllowing from this lowered cocoon production and decreasing number of hatchlings.

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:
other: Acceptable, well documented publication which meets basic scientific principles. Test method described in detail, no GLP and no detailed information on test substance.
Principles of method if other than guideline:
Range finding test and chronic test with Eisenia andrei exposed to different concentrations of Al2(SO4)3 under different pH regimes. Effects followed were survival, growth and cocoon production.
GLP compliance:
no
Analytical monitoring:
yes
Details on sampling:
For 0, 100, and 1000 mg Al/kg dry soil treatments six test containers were prepared in duplicate for analysis of aluminium speciation in soil at three different time intervals (at start, after three weeks exposure, after six weeks exposure).
Details on preparation and application of test substrate:
Artificial soil was prepared following OECD guideline 207 (1984). 70 % quartz sand, 20 % kaolin clay, and 10 % sphagnum peat (sieved with a 1 mm mesh). pH of soil adjusted by adding CaCO3 to soil. Moisture was brought to 55 % (w/w) by adding demineralized water.
Test organisms (species):
other: Eisenia andrei
Animal group:
annelids
Details on test organisms:
TEST ORGANISM
- Common name: earthworm
- Source: own culture on a substrate of horse dung and potting soil
- Age at test initiation (mean and range, SD): adult, 9-12 weeks old, with well-developed clitellum
- Weight at test initiation (mean and range, SD): 259-529 mg (range-finding tests) and 190-489 mg (main test)
Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
no
Total exposure duration:
6 wk
Test temperature:
20 ± 2 °C
pH:
3.5, 5.0 and 7.0 (nominal) (range finding test)
3.5, 4.5, and 6.5 (nominal) (main test)
Moisture:
55 % (w/w)
Details on test conditions:
TEST SYSTEM
- Test container (material, size): 1L glass container
- Amount of soil or substrate: 400 g dry weight
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 1 for each concentration and pH level (range finding test), 3 for each concentration and pH level (main test),
- No. of replicates per control: 1


SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Composition (if artificial substrate): 70 % quartz sand, 20 % kaolin clay, and 10 % sphagnum peat


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


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : survival and cocoon production in range-finding study, additionally weight change


TEST CONCENTRATIONS
- Range finding study
- Test concentrations: 0, 10, 100, 1000, and 4000 mg Al/kg dry soil
Nominal and measured concentrations:
0, 10, 100, 1000, and 4000 mg Al/kg dry soil (range finding study)
0, 10, 32, 100, 320, and 1000 mg Al/kg dry soil (main test)
Reference substance (positive control):
no
Duration:
6 wk
Dose descriptor:
NOEC
Effect conc.:
100 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
Al
Basis for effect:
reproduction
Remarks on result:
other: pH 3.4, 4.3, and 7.3; main study, test phases 2 + 3 = 6 weeks exposure
Duration:
6 wk
Dose descriptor:
NOEC
Effect conc.:
320 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
Al
Basis for effect:
growth
Remarks on result:
other: pH 4.3; main study, test phases 2 + 3 = 6 weeks exposure
Duration:
6 wk
Dose descriptor:
NOEC
Effect conc.:
100 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
Al
Basis for effect:
growth
Remarks on result:
other: pH 3.4 and 7.3; main study, test phases 2 + 3 = 6 weeks exposure
Duration:
6 wk
Dose descriptor:
LC50
Effect conc.:
589 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
Al
Basis for effect:
mortality
Remarks on result:
other: pH 3.4; 95% CL: 555 - 625 (main study)
Duration:
6 wk
Dose descriptor:
EC50
Effect conc.:
189 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
Al
Basis for effect:
growth
Remarks on result:
other: pH 3.4; main study, test phases 2 + 3 = 6 weeks exposure; 95% CL: 18 -2030
Duration:
6 wk
Dose descriptor:
EC50
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
Al
Basis for effect:
growth
Remarks on result:
other: pH 4.3 and 7.3; main study, test phases 2 + 3 = 6 weeks exposure
Duration:
6 wk
Dose descriptor:
EC50
Effect conc.:
294 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
Al
Basis for effect:
reproduction
Remarks on result:
other: pH 3.4; main study, test phases 2 + 3 = 6 weeks exposure; 95% CL: 100 - 868
Duration:
6 wk
Dose descriptor:
EC50
Effect conc.:
529 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
Al
Basis for effect:
reproduction
Remarks on result:
other: pH 4.3; main study, test phases 2 + 3 = 6 weeks exposure; 95% CL: 244 - 1146
Duration:
6 wk
Dose descriptor:
EC50
Effect conc.:
291 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
Al
Basis for effect:
reproduction
Remarks on result:
other: pH 7.3; main study, test phases 2 + 3 = 6 weeks exposure; 95% CL: 131 - 647
Duration:
18 d
Dose descriptor:
LC50
Effect conc.:
457 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
Al
Basis for effect:
mortality
Remarks on result:
other: pH 3.24 ± 0.10; 95% CL: 286 - 729 (range-finding study)
Duration:
18 d
Dose descriptor:
LC50
Effect conc.:
> 4 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
Al
Basis for effect:
mortality
Remarks on result:
other: pH 4.86 ± 0.54 (range-finding study)
Duration:
18 d
Dose descriptor:
LC50
Effect conc.:
> 4 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
Al
Basis for effect:
mortality
Remarks on result:
other: pH 7.22 ± 0.17 (range-finding study)
Duration:
18 d
Dose descriptor:
EC50
Effect conc.:
330 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
Al
Basis for effect:
reproduction
Remarks on result:
other: pH 3.24 ± 0.10 (range-finding study)
Duration:
18 d
Dose descriptor:
EC50
Effect conc.:
197 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
Al
Basis for effect:
reproduction
Remarks on result:
other: pH 4.86 ± 0.54; 95% CL: 8.3 - 4661 (range-finding study)
Duration:
18 d
Dose descriptor:
EC50
Effect conc.:
883 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
Al
Basis for effect:
reproduction
Remarks on result:
other: pH 7.22 ± 0.17; 95% CL: 209 - 3722 (range-finding study)

Table: Effect of Al2(SO4)3 on growth during six week exposure at different pH levels in soil

Concentration mg Al/kg dry soil

% weight change ± SD (n=3) per 10 worms

 

pH 3.4

pH 4.3

pH 7.3

0

44.0 ± 11.1

53.7 ± 2.1

36.7 ± 2.5

10

74.3 ± 8.1

50.7 ± 7.2

36.7 ± 7.0

32

46.7 ± 7.1

56.7 ± 7.1

44.7 ± 10.8

100

42.3 ± 6.0

47.7 ± 4.0

46.0 ± 5.6

320

5.3 ± 30.6*

55.7 ± 1.5

57.7 ± 11.0**

1000

-100 ± 0.0#

39.7 ± 40.0*

49.7 ± 7.0**

# 100% mortality; * significant growth reduction compared to control; ** significant increase in growth compared to control

Table: Effect of Al2(SO4)3 on cocoon production during six week exposure at different pH levels in soil

Concentration mg Al/kg dry soil

Number of cocoons ± SD (n=3) per 10 worms

 

pH 3.4

pH 4.3

pH 7.3

0

7.7 ± 3.1

18.0 ± 5.6

17.0 ± 4.0

10

8.7 ± 3.5

22.3 ± 3.8

22.3 ± 3.8

32

8.7 ± 2.3

16.7 ± 3.2

16.3 ± 5.1

100

9.0 ± 3.5

22.3 ± 5.8

20.3 ± 2.3

320

1.3 ± 2.3*

11.0 ± 2.0*

6.3 ± 4.2*

1000

- #

7.0 ± 3.6*

5.3 ± 5.1*

# 100% mortality; * significant reduction compared to control

Endpoint:
toxicity to soil macroorganisms except arthropods: short-term
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Test method described in detail, no GLP and no detailed information on test substance.
Principles of method if other than guideline:
Range finding test with Eisenia andrei exposed to different concentrations of Al2O3 under different pH regimes. Effect followed was survival.
GLP compliance:
no
Analytical monitoring:
no
Details on preparation and application of test substrate:
Artificial soil was prepared following OECD guideline 207 (1984). 70 % quartz sand, 20 % kaolin clay, and 10 % sphagnum peat (sieved with a 1 mm mesh). pH of soil adjusted by adding CaCO3 to soil. Moisture was brought to 55 % (w/w) by adding demineralized water.
Test organisms (species):
other: Eisenia andrei
Animal group:
annelids
Details on test organisms:
TEST ORGANISM
- Common name: earthworm
- Source: own culture on a substrate of horse dung and potting soil
- Age at test initiation (mean and range, SD): adult, 9-12 weeks old, with well-developed clitellum
- Weight at test initiation (mean and range, SD): 259-529 mg (range-finding test)
Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
no
Total exposure duration:
14 d
Test temperature:
20 ± 2 °C
pH:
2.5, 3.5, 5.0 and 7.0 (nominal)
Moisture:
55 % (w/w)
Details on test conditions:
TEST SYSTEM
- Test container (material, size): 1L glass container
- Amount of soil or substrate: 400 g dry weight
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 1 for each concentration and pH level
- No. of replicates per control: 1

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Composition (if artificial substrate): 70 % quartz sand, 20 % kaolin clay, and 10 % sphagnum peat

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

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : survival

TEST CONCENTRATIONS
- Range finding study
- Test concentrations: 0, 10, 100, 1000 and 5000 mg Al/kg dry soil
Nominal and measured concentrations:
0, 10, 100, 1000 and 5000 mg Al/kg dry soil
Reference substance (positive control):
no
Duration:
14 d
Dose descriptor:
LC50
Effect conc.:
> 5 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
Al
Basis for effect:
mortality
Remarks on result:
other: pH 2.41 ± 0.11
Duration:
14 d
Dose descriptor:
EC50
Effect conc.:
> 5 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
Al
Basis for effect:
mortality
Remarks on result:
other: pH 3.35 ± 0.05
Duration:
14 d
Dose descriptor:
LC50
Effect conc.:
> 5 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
Al
Basis for effect:
mortality
Remarks on result:
other: pH 4.47 ± 0.05
Duration:
14 d
Dose descriptor:
LC50
Effect conc.:
> 5 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
Al
Basis for effect:
mortality
Remarks on result:
other: pH 7.12 ± 0.02
Endpoint:
toxicity to soil macroorganisms except arthropods: short-term
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Acceptable, well documented publication which meets basic scientific principles. Test method described in detail, no GLP and no detailed information on test substance.
Principles of method if other than guideline:
Range finding test with Eisenia andrei exposed to different concentrations of AlCl3 x 6 H2O under different pH regimes. Effect followed was survival.
GLP compliance:
no
Analytical monitoring:
no
Details on preparation and application of test substrate:
Artificial soil was prepared following OECD guideline 207 (1984). 70 % quartz sand, 20 % kaolin clay, and 10 % sphagnum peat (sieved with a 1 mm mesh). pH of soil adjusted by adding CaCO3 to soil. Moisture was brought to 55 % (w/w) by adding demineralized water.
Test organisms (species):
other: Eisenia andrei
Animal group:
annelids
Details on test organisms:
TEST ORGANISM
- Common name: earthworm
- Source: own culture on a substrate of horse dung and potting soil
- Age at test initiation (mean and range, SD): adult, 9-12 weeks old, with well-developed clitellum
- Weight at test initiation (mean and range, SD): 259-529 mg (range-finding test)
Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
no
Total exposure duration:
14 d
Test temperature:
20 ± 2 °C
pH:
3.5, 5.0 and 7.0 (nominal)
Moisture:
55 % (w/w)
Details on test conditions:
TEST SYSTEM
- Test container (material, size): 1L glass container
- Amount of soil or substrate: 400 g dry weight
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 1 for each concentration and pH level
- No. of replicates per control: 1


SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Composition (if artificial substrate): 70 % quartz sand, 20 % kaolin clay, and 10 % sphagnum peat


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


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : survival


TEST CONCENTRATIONS
- Range finding study
- Test concentrations: 0, 10, 100, and 1000 mg Al/kg dry soil
Nominal and measured concentrations:
0, 10, 100, and 1000 mg Al/kg dry soil
Reference substance (positive control):
no
Duration:
14 d
Dose descriptor:
LC50
Effect conc.:
316 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
Al
Basis for effect:
mortality
Remarks on result:
other: pH 3.21 - 3.52 (lowest pH value measured at the highest Al concentration in soil)
Duration:
14 d
Dose descriptor:
LC50
Effect conc.:
> 359 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
Al
Basis for effect:
mortality
Remarks on result:
other: pH 3.69 - 4.38; 95% CL: 274-271 (lowest pH value measured at the highest Al concentration in soil)
Duration:
14 d
Dose descriptor:
EC50
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
Al
Basis for effect:
mortality
Remarks on result:
other: pH 4.37 - 6.70 (lowest pH value measured at the highest Al concentration in soil

Description of key information

Key value for chemical safety assessment

Additional information

There are no studies available on soil macroorganisms other than arthropods for the registered substance reaction mass of aluminium hydroxide and aluminium nitrate and aluminium sulphate.

Nevertheless, aluminium is the most abundant metallic element in the Earth's crust. Based on its ubiquitous occurrence the present natural background concentration far outweighs anthropogenic contributions of aluminium to the terrestrial environment. As detailed in the endpoint summary on terrestrial toxicity in general further toxicity testing on terrestrial organisms is considered unjustified and waiving based on exposure consideration is applied.

However, for reasons of completeness existing data on the terrestrial toxicity of aluminium are provided in addition and summerised here. Based on an analogue approach data are read across to aluminum chloride, aluminum oxide, and aluminum sulphate. Although different aluminium compounds are used as source for testing aluminium,results and concentrations are based on total aluminum and thus facilitate comparison. Van Gestel & Hoogerwerf (2001) conducted three range finding studies with the three aforementioned substances. Worms belonging to the species Eisenia andrei were exposed for 14 days in parallel test series to soils treated with different concentrations of Al and under different pH regimes. For AlCl3 these authors found an increasing toxicity with decreasing pH values with the following LC50 values determined in mg Al/kg soil dw: >1000 at pH 6.70 - 4.37, >359 at pH 4.38 - 3.69, and 316 at pH 3.52 - 3.21. The same trend was found with Al2(SO4)3. Respective LC50 values found are >4000 mg Al/kg soil dw at pH values in the range of 7.39 - 4.32 and 457 mg Al/kg soil dw at pH 3.24. Additionally EC50 values for reproduction were determined for the latter substance. Reproduction was least affected at near neutral pH (7.22 ± 0.17) with an EC50 of 883 mg Al/kg soil dw. Effects were strongest around pH 4.86 with an EC50 of 197 mg Al/kg soil dw. However, at lower pH of 3.24 Al was less toxic to reproduction reflected by the higher EC50 of 330 mg Al/kg soil dw. For Al2O3 no toxicity to earthworms was found at any pH tested (7.14 - 2.30) up to 5000 mg Al/kg soil dw.

A chronic study was carried out by the same authors. Earthworms were exposed for 6 weeks to soils treated with Al2(SO4)3. Survival, growth and cocoon production was followed. Although not as distinct as in the range finding study, a similar pattern of toxicity can be seen with aluminium exhibiting stronger toxicity at pH 3.4 than at higher pH values.As in the range finding study this substance was most toxic at the lowest pH (3.4) and mortality resulted in a LC50 of 589 mg Al/kg soil dw. Effects on growth of worms and production of cocoons did not follow the same trend when exposed to different Al concentrations at different pH. At the lowest pH of 3.4, growth and cocoon production were significantly reduced at 320 mg Al/kg dry soil, while at 1000 mg Al/kg dry soil all earthworms died. At pH 4.3 and 7.3 survival was not affected by 1000 mg Al/kg soil dw. At the same pH and concentration growth and cocoon production was significantly reduced, cocoon production also at a concentration of 320 mg/kg soil dw. At pH 7.3 aluminum affected cocoon production only at 320 and 1000 mg Al/kg dry soil, whereas growth under same conditions was significantly increased, which might be explained by a trade-off between growth and reproduction. The following EC50 and NOEC values (as mg Al/kg soil dw) were determined for these 6 weeks experiments (pH values in parentheses): (a) EC50 growth: >1000 (4.3 and 7.3), 189 (3.4); NOEC growth: 320 (4.3), 100 (3.4 and 7.3), (b) EC50 reproduction: 291 (7.3), 529 (4.3), 294 (3.4); NOEC 100 (3.4, 4.3, and 7.3).

Phillips & Bolger (1998) also used Al2(SO4)3 as test substance and exposed Eisenia fetida for 30 days to different Al concentrations and different pH values. Responses of earthworms to aluminum were followed in terms of survival, growth, cocoon production and cocoon viability. Results obtained show no clear relation between toxicity exhibited by aluminum and pH range. At pH 4.2 the lethal dose was determined to be between 2000 - 4000 mg Al/kg soil. High levels of aluminum inhibited cocoon production at any pH between 4 and 7, whereas intermediate aluminum levels stimulated cocoon production at pH 6 to 7 and low levels of aluminum inhibited hatching and juvenile production at low pH. Cocoon viability in terms of numbers of juveniles hatched from cocoons was stimulated when parents were kept at intermediate levels of aluminum at pH 6 to 7.

A further long-term study is available on AlCl3 by Rundgren & Nilsson (1997). These authors examined effects of different aluminum concentrations and different pH values in soils to Dendrodrilus rubidus in two consecutively conducted tests. This species of earthworm was chosen in order to better reflect natural conditions since Eisenia, as a recommended test species by e.g. OECD guidelines, is not a true soil dwelling organism. At first activity, survival and reproduction was studied in the parent generation exposed for 16 weeks and then hatching, survival and growth in the filial generation exposed for another 48 weeks. In order to determine whether effects were caused by aluminum itself or only by varying pH, a parallel pH series was run without exposure of earthworms to aluminum. It was observed that aluminum had no significant effects on the parameters followed for the parent generation in comparison to controls. For the filial generation, however, responses were observed. Reproduction was significantly affected and a marked drop in reproduction occurred from an average of 12.4 cocoons per surviving adult over the four month period at pH 4.6 to 4.7 cocoons at pH 4.5. A second drop occurred when pH was further lowered from pH 4.5 to 4.2. In the test series with aluminum similar results were obtained. As the concentrations was increased from 100 to 200 mg Al/kg soil, cocoon production decreased from 8.3 to 4.4 cocoons per survivor. Hatchability at 200 mg Al/kg soil significantly decreased. After 26 weeks of incubation all juveniles produced in the first month and maintained in soils of the two highest Al-concentrations had died. Only a few juveniles hatched from cocoons produced in the second month in soils of 200 mg Al/kg soil. All juveniles hatched in soils of 100 mg Al/kg soil (dw) were dead at the end of experiment. In summary, this investigation shows that lowered pH in itself affects D. rubidus and the presence of aluminum does not increase the strength of the response. Lowered pH alone is followed by decreased activity, slightly enhanced weight loss of ageing adult individuals, lowered individual production of cocoons, lowered hatching success of cocoons produced late during the reproductive period, and hampered juvenile growth. Some of these effects might be related, e.g. lowered activity might affect feeding activity and following from this lowered cocoon production and decreasing number of hatchlings.