Aluminium oxide

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

Not reported

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Meets generally accepted scientific standards, well documented and acceptable for assessment

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

Stoneflies collected from a river were exposed to three Al concentrations and a control at pH 7.6 for 96 hours with mortality observed to calculate an LC50.

GLP compliance:

not specified

Specific details on test material used for the study:

- Name of test material: anhydrous AlCl3
- Analytical purity: 99.7%
- Source: MCB Manufacturing Chemists, Inc.
- Lot/batch No.: B7N14

Analytical monitoring:

yes

Details on sampling:

- Concentrations: all test concentrations and controls
- Sampling method: samples were collected before, at the beginning of, and at the end of the exposure period. Samples (15-20 mL) were collected midway between the surface and bottom of the water column in the center of each exposure chamber. A 30 mL adjustable-volume glass-teflon syringe was rinsed three times with the exposure water prior to collecting a sample.
- Sample storage conditions before analysis: All samples were immediately acidified to 1% (v/v) with concentrated nitric acid (Baker Instra analyzed Lot No: 340040). Two mL of potassium chloride solution (95 g KCl/L) was added per 100 mL sample, as suggested in method 202.1. Methods for Chemical Analysis of Water and Wastewater (U.S. EPA, 1979).

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Exposure and control tanks were prepared a minimum of 48 hr in advance of organism introduction to the tanks. The water volume was measured, then added to each exposure tank, and aluminum introduced from a 50,000 mg/l stock solution. Daily adjustments of pH were necessary in many of the exposures. Tests run at pH values above neutrality declined in pH in proportion to the total aluminum present. Controls dropped the least or not at all. Tests run at a pH below neutrality (6.5 nominal) rose in pH in all test chambers.
- Evidence of undissolved material: a precipitate was readily apparent at concentrations of 3.12 mg/L or above
- All other template details: Not reported

Test organisms (species):

other: Acronuria sp.

Details on test organisms:

TEST ORGANISM
- Common name: Stonefly
- Source: collected from the Eau Claire River in Douglas County, WI
- All other template details: Not reported

ACCLIMATION
- Acclimation period: several days
- Acclimation conditions: same as test
- All other template details: Not reported

QUARANTINE (wild caught): Not reported

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

Not reported

Hardness:

47.4 mg/L as CaCO3

Test temperature:

nominal: 16 degrees C
Mean measured water temperature was 16.4 ± 0.10 degrees C (range, 16.3-16.6 degrees C, n=20)

pH:

nominal: 7.6
mean measured pH: 7.46 ± 0.18 (n=22)

Dissolved oxygen:

9.7 ± 0.10 mg/L (n=12)

Salinity:

Not applicable

Nominal and measured concentrations:

Nominal Al (mg/L): 0, 6.25, 12.5, 25.0
Measured Al (mg/L): <0.3, 6.2, 12.3, 22.6

Details on test conditions:

TEST SYSTEM
- Test vessel: glass containers
- Material, size, headspace, fill volume: 5 x 15.5 x 15 cm glass containers (covered with Nytex (R) screen at both ends) and placed into an aquarium containing 12 L of test water.
- Aeration: aquaria were aerated throughout the tests
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- All other template details: Not reported

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Lake Superior water collected from a tap on the City of Cloquet, MN raw water intake in the City of Superior, WI. The source of the water was from approximately 3200 meters offshore on the lake bottom.
- Total solids: 61 ± 5 mg/L
- Particulate matter (total suspended solids): <1.0 mg/L
- Chloride: 1.5 ± 0.42 mg/L
- Alkalinity: 41.6 ± 1.27 mg/L as CaCO3
- Culture medium different from test medium: no
- Intervals of water quality measurement: measured on each batch of water used
- All other template details: Not reported

OTHER TEST CONDITIONS
- Adjustment of pH: Exposure solutions were pH adjusted using either NaOH or HCl of ACS grade purity if the pH differed from the desired test pH by more than 0.2 units.
- Photoperiod: not reported
- Light intensity: not reported

EFFECT PARAMETERS MEASURED: mortality was observed at 24, 48, 72, and 96 hrs

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2 times lower concentration
- All other template details: Not reported

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

> 22.6 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

element

Basis for effect:

mortality

Remarks on result:

other: No mortality at any concentrations

Details on results:

- Mortality of control: 0/5
- All other template details: Not reported

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

Not reported

Not applicable

Conclusions:

There were no mortalities for stoneflies (Acronuria sp.) exposed to Al3+ up to 22.6 mg/L for 96 hours.

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

Not reported

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Meets generally accepted scientific standards, well documented and acceptable for assessment

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

Ceriodaphnia less than 24 hours old were exposed to four Al concentrations and a control at pH 7.5 for 48 hours with mortality observed to calculate an LC50.

GLP compliance:

not specified

Specific details on test material used for the study:

- Name of test material: anhydrous AlCl3
- Analytical purity: 99.7%
- Source: MCB Manufacturing Chemists, Inc.
- Lot/batch No.: B7N14

Analytical monitoring:

yes

Details on sampling:

- Concentrations: all test concentrations and controls
- Sampling method: samples were collected before, at the beginning of, and at the end of the exposure period. Samples (15-20 mL) were collected midway between the surface and bottom of the water column in the center of each exposure chamber. A 30 mL adjustable-volume glass-teflon syringe was rinsed three times with the exposure water prior to collecting a sample.
- Sample storage conditions before analysis: All samples were immediately acidified to 1% (v/v) with concentrated nitric acid (Baker Instra analyzed Lot No: 340040). Two mL of potassium chloride solution (95 g KCl/L) was added per 100 mL sample, as suggested in method 202.1. Methods for Chemical Analysis of Water and Wastewater (U.S. EPA, 1979).

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Exposure and control tanks were prepared a minimum of 48 hr in advance of organism introduction to the tanks. The water volume was measured, then added to each exposure tank, and aluminum introduced from a 50,000 mg/l stock solution. Daily adjustments of pH were necessary in many of the exposures. Tests run at pH values above neutrality declined in pH in proportion to the total aluminum present. Controls dropped the least or not at all. Tests run at a pH below neutrality (6.5 nominal) rose in pH in all test chambers.
- Evidence of undissolved material: a precipitate was readily apparent at concentrations of 3.12 mg/L or above
- All other template details: Not reported

Test organisms (species):

Ceriodaphnia sp.

Details on test organisms:

TEST ORGANISM
- Common name: cladoceran
- Age: less than 24 hours old
- All other template details: Not reported

ACCLIMATION: Not reported

QUARANTINE (wild caught): Not reported

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

Not reported

Hardness:

47.4 mg/L as CaCO3

Test temperature:

23.2 - 25 degrees C

pH:

nominal: 7.5
mean measured pH: 7.68 ± 0.17 (n=10)

Dissolved oxygen:

8.1 - 8.3 mg/L

Salinity:

Not applicable

Nominal and measured concentrations:

Nominal Al (mg/L): 0, 3.12, 6.25, 12.5, 25.0
Measured Al (mg/L): <0.3,

Details on test conditions:

TEST SYSTEM
- Test vessel: beaker
- Material, size, headspace, fill volume: 30 mL beakers with 15 mL of water
- Aeration: test solutions were aerated before the tests for at least 48 hours, but not during the test
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- All other template details: Not reported

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Lake Superior water collected from a tap on the City of Cloquet, MN raw water intake in the City of Superior, WI. The source of the water was from approximately 3200 meters offshore on the lake bottom.
- Total solids: 61 ± 5 mg/L
- Particulate matter (total suspended solids): <1.0 mg/L
- Chloride: 1.5 ± 0.42 mg/L
- Alkalinity: 41.6 ± 1.27 mg/L as CaCO3
- Culture medium different from test medium: no
- Intervals of water quality measurement: measured on each batch of water used
- All other template details: Not reported

OTHER TEST CONDITIONS
- Adjustment of pH: Exposure solutions were pH adjusted using either NaOH or HCl of ACS grade purity if the pH differed from the desired test pH by more than 0.2 units.
- Photoperiod: not reported
- Light intensity: not reported

EFFECT PARAMETERS MEASURED: mortality was observed at 24 and 48 hrs

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2 times lower concentration
- All other template details: Not reported

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

3.69 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

element

Basis for effect:

mortality

Remarks on result:

other: 95% CI: 3.06-4.44 mg/L

Details on results:

- Mortality of control: 0/20
- Complete mortality occurred by 48 hours at 12.4 and 22.8 mg/L Al3+
- All other template details: Not reported

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

Not reported

Not applicable

Conclusions:

The 48 hr LC50 estimate for Ceriodaphnia sp. exposed to Al3+ up to 22.8 mg/L was 3.69 mg/L.

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

Not reported

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Meets generally accepted scientific standards, well documented and acceptable for assessment

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

Scuds collected from a river were exposed to four Al concentrations and a control at pH 7.5 for 96 hours with mortality observed to calculate an LC50.

GLP compliance:

not specified

Specific details on test material used for the study:

- Name of test material: anhydrous AlCl3
- Analytical purity: 99.7%
- Source: MCB Manufacturing Chemists, Inc.
- Lot/batch No.: B7N14

Analytical monitoring:

yes

Details on sampling:

- Concentrations: all test concentrations and controls
- Sampling method: samples were collected before, at the beginning of, and at the end of the exposure period. Samples (15-20 mL) were collected midway between the surface and bottom of the water column in the center of each exposure chamber. A 30 mL adjustable-volume glass-teflon syringe was rinsed three times with the exposure water prior to collecting a sample.
- Sample storage conditions before analysis: All samples were immediately acidified to 1% (v/v) with concentrated nitric acid (Baker Instra analyzed Lot No: 340040). Two mL of potassium chloride solution (95 g KCl/L) was added per 100 mL sample, as suggested in method 202.1. Methods for Chemical Analysis of Water and Wastewater (U.S. EPA, 1979).

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Exposure and control tanks were prepared a minimum of 48 hr in advance of organism introduction to the tanks. The water volume was measured, then added to each exposure tank, and aluminum introduced from a 50,000 mg/l stock solution. Daily adjustments of pH were necessary in many of the exposures. Tests run at pH values above neutrality declined in pH in proportion to the total aluminum present. Controls dropped the least or not at all. Tests run at a pH below neutrality (6.5 nominal) rose in pH in all test chambers.
- Evidence of undissolved material: a precipitate was readily apparent at concentrations of 3.12 mg/L or above
- All other template details: Not reported

Test organisms (species):

Gammarus sp.

Details on test organisms:

TEST ORGANISM
- Common name: Scud, Gammarus pseudolimnaeus
- Source: collected from the Eau Claire River in Douglas County, WI
- Feeding during test
- Food type: aspen (Populus tremuloides) aged leaf disc
- Amount: 1 leaf disk 1.5 cm in diameter
- Frequency: added once before test
- All other template details: Not reported

ACCLIMATION
- Acclimation period: several weeks
- Acclimation conditions: same as test
- All other template details: Not reported

QUARANTINE (wild caught): Not reported

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

Not reported

Hardness:

47.4 mg/L as CaCO3

Test temperature:

nominal: 18 degrees C.
Mean measured water temperature was 18.1 ± 0.48 degrees C (range, 17.4-18.7 degrees C)

pH:

nominal: 7.5
mean measured pH: 7.53 ± 0.07 (n=29)

Dissolved oxygen:

9.4 ± 0.28 mg/L (n=20)

Salinity:

Not applicable

Nominal and measured concentrations:

Nominal Al (mg/L): 0, 3.12, 6.25, 12.5, 25.0
Measured Al (mg/L): <0.3, 2.6, 5.9, 12.4, 24.1

Details on test conditions:

TEST SYSTEM
- Test vessel: beaker
- Material, size, headspace, fill volume: 250 mL glass beaker. Beakers had contained two opposing 2.5 cm diameter holes in the sides covered with Nytex (R) screen. Nine beakers per exposure concentration were suspended in a 30.5 x 21 x 25.5 cm aquarium containing 12 L of test water.
- Aeration: aquaria were aerated throughout the tests
- No. of organisms per vessel: 9
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- All other template details: Not reported

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Lake Superior water collected from a tap on the City of Cloquet, MN raw water intake in the City of Superior, WI. The source of the water was from approximately 3200 meters offshore on the lake bottom.
- Total solids: 61 ± 5 mg/L
- Particulate matter (total suspended solids): <1.0 mg/L
- Chloride: 1.5 ± 0.42 mg/L
- Alkalinity: 41.6 ± 1.27 mg/L as CaCO3
- Culture medium different from test medium: no
- Intervals of water quality measurement: measured on each batch of water used
- All other template details: Not reported

OTHER TEST CONDITIONS
- Adjustment of pH: Exposure solutions were pH adjusted using either NaOH or HCl of ACS grade purity if the pH differed from the desired test pH by more than 0.2 units.
- Photoperiod: not reported
- Light intensity: not reported

EFFECT PARAMETERS MEASURED: mortality was observed at 24, 48, 72, and 96 hrs

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2 times lower concentration
- All other template details: Not reported

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

22 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

element

Basis for effect:

mortality

Remarks on result:

other: 95% CI: 13.8-35.2 mg/L

Details on results:

- Mortality of control: 0/9
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: A considerable amount of floc occurred at the bottoms of the beakers, particularly at the three highest concentrations. The leaf discs were often situated in the floc with scuds present under the discs. Thus, many of the scuds survived even while apparently spending considerable amounts of time in the Al floc.
- All other template details: Not reported

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

Not reported

Not applicable

Conclusions:

The 96 hr LC50 estimate for scuds (Gammarus pseudolimnaeus) exposed to Al3+ up to 24.1 mg/L was 22.0 mg/L.

Reference 4

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

Not reported

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Meets generally accepted scientific standards, well documented and acceptable for assessment

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

Snails were exposed to three Al concentrations and a control at pH 6.5, 7.5, and 8.5 for 96 hours with mortality observed to calculate an LC50.

GLP compliance:

not specified

Specific details on test material used for the study:

- Name of test material: anhydrous AlCl3
- Analytical purity: 99.7%
- Source: MCB Manufacturing Chemists, Inc.
- Lot/batch No.: B7N14

Analytical monitoring:

yes

Details on sampling:

- Concentrations: all test concentrations and controls
- Sampling method: samples were collected before, at the beginning of, and at the end of the exposure period. Samples (15-20 mL) were collected midway between the surface and bottom of the water column in the center of each exposure chamber. A 30 mL adjustable-volume glass-teflon syringe was rinsed three times with the exposure water prior to collecting a sample.
- Sample storage conditions before analysis: All samples were immediately acidified to 1% (v/v) with concentrated nitric acid (Baker Instra analyzed Lot No: 340040). Two mL of potassium chloride solution (95 g KCl/L) was added per 100 mL sample, as suggested in method 202.1. Methods for Chemical Analysis of Water and Wastewater (U.S. EPA, 1979).

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Exposure and control tanks were prepared a minimum of 48 hr in advance of organism introduction to the tanks. The water volume was measured, then added to each exposure tank, and aluminum introduced from a 50,000 mg/l stock solution. Daily adjustments of pH were necessary in many of the exposures. Tests run at pH values above neutrality declined in pH in proportion to the total aluminum present. Controls dropped the least or not at all. Tests run at a pH below neutrality (6.5 nominal) rose in pH in all test chambers.
- Evidence of undissolved material: a precipitate was readily apparent at concentrations of 3.12 mg/L or above
- All other template details: Not reported

Test organisms (species):

other: Physa sp.

Details on test organisms:

TEST ORGANISM
- Common name: Snail
- Source: received from Fattig Fish Hatchery, Brady, NE
- Weight: for pH 7.5 test, average weight was 0.033 ± 0.013 g; pH 6.5 and 8.5 tests, average weight was 0.9 ± 0.02 g
- All other template details: Not reported

ACCLIMATION
- Acclimation period: several days
- Acclimation conditions: same as test
- All other template details: Not reported

QUARANTINE (wild caught): Not reported

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

Not reported

Hardness:

47.4 mg/L as CaCO3

Test temperature:

pH 7.5 Nominal: 22 degrees C.
pH 7.5 Mean measured water temperature was 22.2 ± 4 degrees C (range, 20.9-22.6 degrees C, n=50)
pH 6.5 and 8.5 nominal: 17 degrees C
pH 6.5 measured (mean and SD (range)): 16.9 ± 0.69 (15.9-17.8) degrees C
pH 8.5 measured (mean and SD (range)): 17.5 ± 0.70 (16.0-l8.5) degrees C

pH:

pH nominal: 6.5, 7.5, 8.5
pH measured: 6.59 ± 0.15, 7.55 ± 0.13, 8.17 ± 0.42

Dissolved oxygen:

pH 6.5 (mg/L): 8.7 ± 0.64
pH 7.5 (mg/L): 8.1 ± 0.5
pH 8.5 (mg/L): 8.6 ± 0.22

Salinity:

Not applicable

Nominal and measured concentrations:

pH 7.5 Nominal Al (mg/L): 0, 6.25, 12.5, 50.0
pH 7.5 Measured Al (mg/L) for both replicates: <0.3/<0.3, 6.4/6.3, 12.3/12.3, 24.8/25.2, 50.2/50.4
pH 6.5 and 8.5 Nominal Al (mg/L): 0, 3.12, 6.25, 12.5, 25
pH 6.5 Measured Al (mg/L) for both replicates: <0.3/<0.3, 2.4/2.6, 5.2/5.6, 12.0/11.6, 23.4/23.5
pH 8.5 Measured Al (mg/L) for both replicates: <0.3/<0.3, 3.1/3.5, 6.0/6.4, 12.1/12.0, 12.4/24.7

Details on test conditions:

TEST SYSTEM
- Test vessel: stainless steel mesh wire cylinders
- Material, size, headspace, fill volume: stainless steel mesh wire cylinders (8-10 cm length x 3 cm diameter) closed at either end with neoprene stoppers which had been pre-soaked in laboratory water for 2-3 days. The cylinders were laid horizontally on the floors of glass aquaria containing 12 L of water.
- Aeration: aquaria were aerated throughout the tests
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- All other template details: Not reported

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Lake Superior water collected from a tap on the City of Cloquet, MN raw water intake in the City of Superior, WI. The source of the water was from approximately 3200 meters offshore on the lake bottom.
- Total solids: 61 ± 5 mg/L
- Particulate matter (total suspended solids): <1.0 mg/L
- Chloride: 1.5 ± 0.42 mg/L
- Alkalinity: 41.6 ± 1.27 mg/L as CaCO3
- Culture medium different from test medium: no
- Intervals of water quality measurement: measured on each batch of water used
- All other template details: Not reported

OTHER TEST CONDITIONS
- Adjustment of pH: Exposure solutions were pH adjusted using either NaOH or HCl of ACS grade purity if the pH differed from the desired test pH by more than 0.2 units.
- Photoperiod: not reported
- Light intensity: not reported

EFFECT PARAMETERS MEASURED: mortality was observed at 96 hrs

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2 times lower concentration
- All other template details: Not reported

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

30.6 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

element

Basis for effect:

mortality

Remarks on result:

other: pH 7.5, 95% CI: 23.1-40.6 mg/L

Details on results:

- Mortality of control: 0/30
- Insufficient mortalities were observed at pH 6.5 and 8.5 to calculate LC50 (see table B4), no other statistics were performed
- All other template details: Not reported

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

Not reported

Table B4. Mortalities of Snails (Physa sp.) Exposed to Al3+ for 96 hr in Lake Superior Water at pH 6.59, 7.55, and 8.17.

 

pH 6.59	Al 3+ concentrations (mg/L)
Nominal	0.0	0.0	3.12	3.12	6.25	6.25	12.5	12.5	25	25
Measured	<0.3	<0.3	2.4	2.6	5.2	5.6	12.0	11.6	23.4	23.5
96hr mortalities	0	0	0	0	0	0	4	1	1	0
pH 7.55	Al 3+ concentrations (mg/L)
Nominal	0.0	0.0	6.25	6.25	12.5	12.5	25	25	50	50
Measured	<0.3	<0.3	6.4	6.3	12.3	12.3	24.8	25.2	50.2	50.4
96hr mortalities	0	0	0	0	0	0	3	0	4	5
pH 8.17	Al 3+ concentrations (mg/L)
Nominal	0.0	0.0	3.12	3.12	6.25	6.25	12.5	12.5	25	25
Measured	<0.3	<0.3	3.1	3.5	6.0	6.4	12.1	12.0	12.4	24.7
96hr mortalities	0	0	0	0	0	0	1	1	0	0

 

Conclusions:

The 96 hr LC50 estimate for snails (Physa sp.) exposed to Al3+ up to 50.3 mg/L was 30.6 mg/L. Insufficient mortalities occurred at pH 6.5 and 8.5 for LC50 calculations.

Reference 5

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

Not reported

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Meets generally accepted scientific standards, well documented and acceptable for assessment

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

other: USEPA, method 2012.0 D. magna acute toxicity tests with effluents and receiving waters in EPA-821-R-02-012 "Methods for Measuring the Acute Toxicity of Effluents and Receiving Water to Freshwater Organisms and Marine Organisms."

Deviations:

not specified

Principles of method if other than guideline:

Not applicable

GLP compliance:

no

Remarks:

The testing laboratory is operating according to GLP specifications but is not cerfied according to GLP.

Specific details on test material used for the study:

- Molecular formula: Al(NO3)3 x 9H2O

Analytical monitoring:

yes

Details on sampling:

- Concentrations: control and all test concentrations
- Sampling method: not reported
- Sample storage conditions before analysis: the extraction aliquot can be stored at 4 degrees C in an Ependorf tube

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Al was dissolved in standard laboratory media with synthetic buffers to reduce pH drift (MES, pK 6.1, for pH 6.0; MOPS, pK 7.2 for pH 7.0; HEPES, pK 7.5 for pH 8.0)
- All other template details: Not reported

Test organisms (species):

Ceriodaphnia dubia

Details on test organisms:

TEST ORGANISM
- Age at study initiation: less than 24-hours old
- Feeding during test: none
- All other template details: Not reported

ACCLIMATION: Not reported

QUARANTINE: Not reported

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

Not reported

Hardness:

Nominal: 10.6, 60, 120 mg/L

Test temperature:

20 ± 1 degree C

pH:

Nominal: 6,7,8
Measured: see Tables 1-3 for measured values

Dissolved oxygen:

Not reported

Salinity:

Not reported

Nominal and measured concentrations:

Nominal (μg/L): 0,62.5-15000 μg/L, depending on the test
Measured: see Tables 1-3 for measured values

Details on test conditions:

TEST SYSTEM
- Material, size, headspace, fill volume: 30 mL plastic beakers filled with 25 mL of test solution
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- All other template details: Not reported

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: EPA very soft water
- Alkalinity: 7 mg/L
- Ca/mg ratio: 1:1
- NaHCO3: 12.0 mg/L
- All other template details: Not reported

OTHER TEST CONDITIONS
- Adjustment of pH: not reported
- Photoperiod: 16 hr light:8 hr dark
- Light intensity: 10-20 μE/m2/s (50-100 ft candles)

EFFECT PARAMETERS MEASURED: mortality measured at 48 hrs

TEST CONCENTRATIONS
- Spacing factor for test concentrations: linear series
- All other template details: Not reported

Reference substance (positive control):

not specified

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

13.5 µg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

dissolved

Basis for effect:

mortality

Remarks on result:

other: pH 6, Hardness 10.6 mg/L, DOC 0 mg/L; Nominal 71.12 µg/L

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

6.8 µg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

dissolved

Basis for effect:

mortality

Remarks on result:

other: pH 6, Hardness 60 mg/L, DOC 0 mg/L; Nominal 110.8 µg/L

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

11.4 µg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

dissolved

Basis for effect:

mortality

Remarks on result:

other: pH 6, Hardness 120 mg/L, DOC 0 mg/L; Nominal 2007.7 µg/L

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

22.2 µg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

dissolved

Basis for effect:

mortality

Remarks on result:

other: pH 6, Hardness 10.6 mg/L, DOC 2 mg/L; Nominal 686.5 µg/L

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

14.4 µg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

dissolved

Basis for effect:

mortality

Remarks on result:

other: pH 6, Hardness 60 mg/L, DOC 2 mg/L; Nominal 1137.1 µg/L

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

5.7 µg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

dissolved

Basis for effect:

mortality

Remarks on result:

other: pH 6, Hardness 120 mg/L, DOC 2 mg/L; Nominal 3386.8 µg/L

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

12.7 µg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

dissolved

Basis for effect:

mortality

Remarks on result:

other: pH 6, Hardness 10.6 mg/L, DOC 4 mg/L; Nominal 1558.1 µg/L

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

32.1 µg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

dissolved

Basis for effect:

mortality

Remarks on result:

other: pH 6, Hardness 60 mg/L, DOC 4 mg/L; Nominal 8046.7 µg/L

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

52 µg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

dissolved

Basis for effect:

mortality

Remarks on result:

other: pH 6, Hardness 120 mg/L, DOC 4 mg/L; Nominal 10484.2 µg/L

Details on results:

- There were insufficient mortalities in all of the pH 7 and 8 tests for an LC50 to be determined. No other statistics to determine effect levels were performed.
- Mortality of control: maximum was 5% (n=1) mortality of control in all tests
- Effect concentrations exceeding solubility of substance in test medium: see Tables 1-3 for concentrations exceeding solubility
- All other template details: Not reported

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

LC50 values, with a 95% confidence interval, were calculated from the experimental data using the Probit method (EPA) and ICp software v2.0 (EPA). The EC50 was calculated for “Nominal Al concentrations” using the software already mentioned, but in the case of “Dissolved Al” and “Monomeric Al” concentrations, the EC50 were determined by a simple proportionality calculation based in the Nominal Al concentration values closest to the EC50 or IC50.

Table 3. Acute Al toxicity with Ceriodaphnia dubia in EPA Very Soft water (Hardness 10.6,60,120 mg/L as CaCO3) at pH 8 and DOC of 0,2,4 mg/L.

pH	Hardness (mg/L)	DOC (mg/L)	Nominal (µg/L)	Mean Dissolved (µg/L)	Mean pH
8	10.6	0	0.0	0.0	7.93
			1000	284.9	7.90
			2000	287.3	7.87
			3000	272.3	7.84
			4000	258.2	7.80
			5000	242.7	7.77
8	60	0	0.0	0.0	7.92
			1000	429.5	7.88
			2000	381.4	7.84
			3000*	370.4	7.81
			4000*	313.7	7.77
			5000*	291.7	7.74
8	120	0	0.0	0.0	7.95
			1000	374.4	7.93
			2000*	312.8	7.90
			3000*	278.9	7.86
			4000*	252.7	7.84
			5000*	218.0	7.80
8	10.6	2	0.0	0.0	8.14
			1000	337.7	8.09
			3000*	265.2	7.98
			5000*	233.8	7.88
			10000*	139.6	7.69
			15000*	82.7	7.49
8	60	2	0.0	0.0	7.92
			1000	616.9	7.87
			3000	636.9	7.80
			5000*	323.7	7.74
			10000*	225.2	7.58
			15000*	163.3	7.41
8	120	2	0.0	0.0	7.95
			1000	367.1	7.92
			3000*	294.8	7.86
			5000*	228.8	7.77
			10000*	181.2	7.63
			15000*	184.6	7.46
8	10.6	4	0.0	0.0	7.93
			1000	386.1	7.89
			3000	264.2	7.83
			5000*	181.6	7.75
			10000*	115.6	7.59
			15000*	52.7	7.39
8	60	4	0.0	0.0	7.89
			1000	816.6	7.85
			3000	601.6	7.78
			5000	404.0	7.71
			10000	210.1	7.53
			15000	141.8	7.35
8	120	4	0.0	0.0	7.87
			1000	246.2	7.83
			3000	247.0	7.75
			5000	250.8	7.67
			10000*	287.0	7.50
			15000*	408.7	7.32

* - Precipitation observed in the vessel

Conclusions:

The 48-hr LC50 values for Ceriodaphnia dubia exposed to Al as AlNO3 could not be calculated for all tests at pH 7 and 8. The 48-hr LC50 for average dissolved concentrations at pH 6 ranged from 5.7 - 52 µg/L depending on the hardness and dissolved organic carbon.

Reference 6

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

Not reported

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Meets generally accepted scientific standards, well documented and acceptable for assessment

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

other: USEPA, method 2012.0 D. magna acute toxicity tests with effluents and receiving waters in EPA-821-R-02-012 "Methods for Measuring the Acute Toxicity of Effluents and Receiving Water to Freshwater Organisms and Marine Organisms."

Deviations:

not specified

Principles of method if other than guideline:

Not applicable

GLP compliance:

no

Remarks:

The testing laboratory is operating according to GLP specifications but is not cerfied according to GLP.

Specific details on test material used for the study:

- Molecular formula: Al(NO3)3 x 9H2O

Analytical monitoring:

yes

Details on sampling:

- Concentrations: control and all test concentrations
- Sampling method: not reported
- Sample storage conditions before analysis: the extraction aliquot can be stored at 4 degrees C in an Ependorf tube

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Al was dissolved in standard laboratory media with synthetic buffers to reduce pH drift (MES, pK 6.1, for pH 6.0; MOPS, pK 7.2 for pH 7.0; HEPES, pK 7.5 for pH 8.0)
- All other template details: Not reported

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Age at study initiation: less than 24-hours old
- Feeding during test: none
- All other template details: Not reported

ACCLIMATION: Not reported

QUARANTINE: Not reported

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

Not reported

Hardness:

168 mg/L

Test temperature:

20 ± 1 degree C

pH:

Nominal: 6,7,8

Dissolved oxygen:

Not reported

Salinity:

Not reported

Nominal and measured concentrations:

Nominal (μg/L): 0,100,200,300,400,500
Nominal for pH 8 with 0,2,3 mg/L DOC (μg/L): 0,400,600,800,1000,1200
Measured: see Tables 1 and 2

Details on test conditions:

TEST SYSTEM
- Material, size, headspace, fill volume: 30 mL test chamber filled with 25 mL of test solution
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- All other template details: Not reported

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: low alkalinity EPA hard water
- Alkalinity: 11 mg/L
- Ca/mg ratio: 120:120
- NaHCO3: 19.2 mg/L
- All other template details: Not reported

OTHER TEST CONDITIONS
- Adjustment of pH: not reported
- Photoperiod: 16 hr light:8 hr dark
- Light intensity: 10-20 μE/m2/s (50-100 ft candles)

EFFECT PARAMETERS MEASURED: mortality measured at 48 hrs

TEST CONCENTRATIONS
- Spacing factor for test concentrations: linear series
- All other template details: Not reported

Reference substance (positive control):

not specified

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

>= 15.2 µg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

dissolved

Basis for effect:

mortality

Remarks on result:

other: pH 6

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

>= 18.8 µg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

dissolved

Basis for effect:

mortality

Remarks on result:

other: pH 7

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

414.6 µg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

dissolved

Basis for effect:

mortality

Remarks on result:

other: pH 8, 95% CI 400.8-422.8 µg/L

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

389.5 µg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

dissolved

Basis for effect:

mortality

Remarks on result:

other: pH 8, DOC 0 mg/L, 95% CI 377.9-400.2 µg/L

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

>= 504.8 µg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

dissolved

Basis for effect:

mortality

Remarks on result:

other: pH 8, DOC 2 mg/L

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

>= 671.2 µg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

dissolved

Basis for effect:

mortality

Remarks on result:

other: pH 8, DOC 3 mg/L

Details on results:

- Mortality of control: 0% mortality of control in all tests
- All other template details: Not reported

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

LC50 values, with a 95% confidence interval, were calculated from the experimental data using the Probit method (EPA) and ICp software v2.0 (EPA). The EC50 was calculated for “Nominal Al concentrations” using the software already mentioned, but in the case of “Dissolved Al” and “Monomeric Al” concentrations, the EC50 were determined by a simple proportionality calculation based in the Nominal Al concentration values closest to the EC50 or IC50.

Not applicable

Conclusions:

The 48-hr LC50 values for Daphnia magna exposed to Al as AlNO3 were 414.6 and 389.5 µg dissolved Al/L for two tests at pH 8 and DOC 0 mg/L. The LC50 values could not be calculated for pH 6 and 7, as well as pH 8 with DOC of 2 and 3 mg/L due to insufficient mortality at the concentrations tested (up to 500 µg/L nominal for pH 6 and 7, and 1200 µg/L nominal for pH 8 tests).

Reference 7

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

May 1992

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

other: USEPA 1985. Methods for measuring the acute toxicity of effluents to freshwater and marine organisms. EPA/600/4-85/013.

Deviations:

not specified

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes

Specific details on test material used for the study:

- Name of test material: aluminum chloride hexahydrate
- Molecular formula: AlCl3 x 6H2O
- Source: Sigma Chemical Company, St. Louis, Missouri, USA

Analytical monitoring:

yes

Details on sampling:

- Concentrations: each test treatment and controls at test initiation and at test termination (or on the day that 100% mortality occurred in any chamber)
- Sampling method: test solutions from all test chambers within each treatment were composited
- Sample storage conditions before analysis: samples were immediately acidified with 1 mL nitric acid

Vehicle:

no

Details on test solutions:

Not reported

Test organisms (species):

Ceriodaphnia dubia

Details on test organisms:

TEST ORGANISM
- Source: from the FCETL in-house cultures
- Age at study initiation: < 24 hours old
- Feeding during test: none
- All other template details: Not reported

ACCLIMATION: Not reported

QUARANTINE (wild caught): Not applicable

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

Not reported

Hardness:

Nominal: 25, 50, 100, 200 mg/L as CaCO3
Measured: 26, 46, 96, 194 mg/L as CaCO3

Test temperature:

20 ± 2 degrees C

pH:

pH: 7.5, 7.6, 7.8, 8.1 for tests with hardness 25, 50, 100, 200 mg/L

Dissolved oxygen:

greater than 6.4 mg/L (84% saturation)

Salinity:

Not reported

Nominal and measured concentrations:

Nominal with 25 mg/L hardness (mg/L): 0, 0.6, 1.25, 2.5, 5, 10
Nominal with 50 mg/L hardness (mg/L): 0, 1.25, 2.5, 5, 10, 20
Nominal with 100 mg/L hardness (mg/L): 0, 3.125, 6.25, 12.5, 25, 50
Nominal with 200 mg/L hardness (mg/L): 0, 6.25, 12.5, 25, 50, 100
Measured values in Table 1 and 2

Details on test conditions:

TEST SYSTEM
- Test vessel: 100 mL glass beakers
- Material, size, headspace, fill volume: 100 mL glass beakers containing 50 mL test solution
- Aeration: none
- Renewal rate of test solution (frequency/flow rate): renewed at 24 hours
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- All other template details: Not reported

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted water according to USEPA 1985 guidelines
- Chlorine (mg/L): <0.05 in all tests
- Alkalinity: 14, 30, 62, 134 mg/L as CaCO3 for hardness values 25,50,100,200 - respectively
- Conductivity (µS/cm): 90, 169, 330, 649 for hardness values 25,50,100,200 - respectively
- All other template details: Not reported

OTHER TEST CONDITIONS
- Adjustment of pH: not reported
- Photoperiod: 16 hrs light: 8 hrs dark
- Light intensity: fluorescent lighting

EFFECT PARAMETERS MEASURED: mortality at 24 and 48 hrs

TEST CONCENTRATIONS
- Spacing factor for test concentrations: linear
- All other template details: Not reported

Reference substance (positive control):

yes

Remarks:

NaCl

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

0.72 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

element

Basis for effect:

mortality

Remarks on result:

other: 25 mg/L hardness; pH 7.5

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

1.88 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

element

Basis for effect:

mortality

Remarks on result:

other: 50 mg/L hardness; pH 7.6

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

2.45 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

element

Basis for effect:

mortality

Remarks on result:

other: 100 mg/L hardness; pH 7.8

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 99.6 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

element

Basis for effect:

mortality

Remarks on result:

other: 200 mg/L hardness; pH 8.1

Details on results:

- Mortality of control: none
- All other template details: Not reported

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- Mortality: yes
- EC50/LC50: 24-hr LC50 1,470 mg/L Cl- as calculated by the binomial method
- Other: not reported

Reported statistics and error estimates:

LC50 values were calculated for each test using a computer program supplied by USEPA (1985).

Not applicable

Conclusions:

The 48-hr LC50 for Ceriodaphnia dubia exposed to Aluminium chloride increased from 0.72 to greater than 99.6 mg/L with water hardness increasing from 25 to 200 mg/L.

Reference 8

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

Not reported

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Meets generally accepted scientific standards, well documented and acceptable for assessment

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

Acute toxicity tests were conducted as static tests in constant temperature water baths for 48 hr. Each beaker held 10 neonate daphnia that were 12 ± 12 hr old at the start of the test. Two of the four replicates were not fed. Survival was observed daily.

GLP compliance:

not specified

Specific details on test material used for the study:

- Molecular formula: (Al)2(SO4)3 x 16H2O
- Analytical purity: reagent grade

Analytical monitoring:

yes

Details on sampling:

- Concentrations: all concentrations
- Sampling method: samples were taken at the beginning and end of each test. Samples were drawn into 15 ml tubes and acidified with 1 drop of 1.0 N HNO3.
- Sample storage conditions before analysis: Not reported

Vehicle:

not specified

Details on test solutions:

Not reported

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: daphnid
- Source: commercially available source, Aquatic Life Company, Minneapolis, MN.
- Age at study initiation: neonates 12 ± 12 hrs old
- Method of breeding: used a series of 250 ml beakers containing 200 ml water with food at a concentration of 30 mg/L in each beaker. Water was totally renewed three times per week with approximately 5 adult daphnia per beaker transferred to the new solution.
- Feeding during test
- Amount: 30 mg/l with an eye dropper
- All other template details: Not reported

ACCLIMATION: Not reported

QUARANTINE (wild caught): Not applicable

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

Not reported

Hardness:

Hard, value not reported

Test temperature:

20.1 degrees C (overall mean)

pH:

7.05 (overall mean)

Dissolved oxygen:

8.08 mg/L (overall mean)

Salinity:

Not reported

Nominal and measured concentrations:

Range of treatment means (mg/L): 4.0-43.0 total, 1.5-3.0 supernatant

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 250 ml beakers containing 200 ml solution
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2
- All other template details: Not reported or Not applicable

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: well water tapping the Jordan sandstone stratum underlying the Minneapolis-St. Paul metropolitan area, with iron catalytically removed
- Culture medium different from test medium: no
- Intervals of water quality measurement: beginning and end of experiments
- All other template details: Not reported

OTHER TEST CONDITIONS: Not reported

EFFECT PARAMETERS MEASURED: survival observed daily

TEST CONCENTRATIONS: Not reported

Reference substance (positive control):

not specified

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

38.2 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

element

Basis for effect:

mortality

Remarks on result:

other: all replicates, with and without food

Details on results:

No details provided

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

LC50 values were derived by a computer program that calculates LC50 values by four methods. The method that gave the tightest confidence interval was used.

Not applicable

Conclusions:

Daphnia magna exposed to aluminium sulphate for 48 hr had an LC50 38.2 mg/L, regardless if the daphnids were fed or not. An LC50 could not be calculated at 96 hr.

Reference 9

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

disregarded due to major methodological deficiencies

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

other: Significant methodological deficiencies: statistics were not used to determine effect levels

Specific details on test material used for the study:

- Molecular formula: (Al)2(SO4)3 x 18H2O
- Physical state: crystals
- Analytical purity: reagent grade

Test organisms (species):

other aquatic arthropod: Tanytarsus dissimilis

Reference 10

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

Not reported

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Meets generally accepted scientific standards, well documented and acceptable for assessment

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

Acute exposures were conducted with four replicates for each Al concentration and control. Five C. dubia neonates less than 24 hours old were added to a beaker via a fire-polished, wide-mouth pipette and observed for 48 hours. Immobilisation was used as the endpoint. Organisms were not fed during the exposures.

GLP compliance:

not specified

Specific details on test material used for the study:

- Name of test material: aluminum chloride
- Source: MCB Chemicals
- Analytical purity: 99.8 %
- Lot/batch No.: B7N14

Analytical monitoring:

yes

Details on sampling:

- Concentrations: all concentrations
- Sampling method: samples were collected at the initiation, prior to and after solution renewal, and at the termination of the tests
- Sample storage conditions before analysis: samples were immediately acidified with HNO3 to 0.1% (v/v, pH range 1.5 to 2.0), mixed, and allowed to stand (>1 hr) before aliquots were withdrawn and composited.

Vehicle:

not specified

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: aliquots of 5000 mg/L Al stock was added to filtered raw Lake Superior or dechlorinated tap water.
- All other template details: Not reported

Test organisms (species):

Ceriodaphnia dubia

Details on test organisms:

TEST ORGANISM
- Common name: cladoceran
- Source: initially from USEPA Research Laboratory, Duluth, MN stock culture - then cultured at the University of Wisconsin-Superior culture facility for several generations before use in these tests
- Age at study initiation: less than 24 hours old
- Method of breeding: adult cladocerans were cultured individually in 30 mL containers and were transferred to new water every Monday, Wednesday, and Friday. They were fed a 1:1:1 ratio of fermented trout chow solution, yeast solution, and green algae (Selenastrum capricornutum) one daily.
- Feeding during test: None
- All other template details: Not reported

ACCLIMATION: none

QUARANTINE: Not applicable

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

Not reported

Hardness:

50-50.5 mg CaCO3/L

Test temperature:

25 ± 2 degrees C

pH:

Nominal: 7.2-7.4, 7.6-7.9, 8.1,
Measured: 7.42 ± 0.02, 7.86 ± 0.04, 8.13 ± 0.03,

Dissolved oxygen:

Measured DO for the 3 pH levels (mg/L): 7.0 ± 0.2, N/A (temperature reported), 7.4 ± 0.1

Salinity:

Not applicable

Nominal and measured concentrations:

Measured in pH 7.42 test (mg/L): control, 1.5, 2.6, 5.0, 9.6, 16.2
Measured in pH 7.86 test (mg/L): control, 1.2, 2.2, 6.0, 10.6, 18.6 (or 20.6 from text)
Measured in pH 8.13 test (mg/L): control, 1.3, 2.1, 3.7, 8.5, 17.7

Details on test conditions:

TEST SYSTEM
- Test vessel: 30 mL beakers
- Material, size, headspace, fill volume: 30 mL containers with 16 mL of test solution
- Aeration: test solutions were aerated for at least 24 hours before test initiation, none during testing
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- All other template details: Not reported

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dilution water was either raw Lake Superior water for pH 7.42 and 8.1 or UW-Superior Laboratory water for pH 7.86
- Total solids (mg/L at 105 degrees C): 80, 70
- Particulate matter (suspended solids, mg/L): 2, <1
- Chloride (mg Cl/L): 2.8, 3.8
- Alkalinity (mg CaCO3/L): 46.0, 39.6
- All other measured characteristics presented in Table 1
- All other template details: Not reported

OTHER TEST CONDITIONS
- Adjustment of pH: addition of reagent grade HCl or NaOH prior to test initiation
- Photoperiod: not reported
- Light intensity: not reported

EFFECT PARAMETERS MEASURED: immobilisation at 48 hours

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2x
- All other template details: Not reported

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

1.9 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

other: acid exchangeable

Basis for effect:

other: immobilisation

Remarks on result:

other: pH 7.42; 95% CI: 1.78-2.04 mg/L

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

1.5 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

other: acid exchangeable

Basis for effect:

other: immobilisation

Remarks on result:

other: pH 7.86; 95% CI not reliable

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

2.56 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

other: acid exchangeable

Basis for effect:

other: immobilisation

Remarks on result:

other: pH 8.13; 95% CI 2.14-3.06 mg/L

Details on results:

- Mortality of control: no control animals died
- All other template details: Not reported

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

Estimates of the 48-hr EC50 Al concentration and the 95% confidence limits were made using the Trimmed spearman-Karber method (Hamilton et al. 1977). Replicate exposures were combined to determine the EC50 estimates.

Table 2. Mean cumulative percent survival of Ceriodaphnia dubia exposed to aluminium at 3 pH levels.

	Aluminium Exposure (mg/L)
pH level	Control	1.5	2.6	5.0	9.6	16.2
7.42	100	85	5	0	0	0
	Control	1.2	2.2	6.0	10.6	18.6
7.86	100	75	0	0	0	0
	Control	1.3	2.1	3.7	8.5	17.7
8.1	100	100	50	25	0	0

Conclusions:

The 48-hr EC50s for immobilisation of C.dubia exposed to aluminium in waters of pH 7.42, 7.86, and 8.13 were 1.9, 1.5, and 2.6 mg/L, respectively.

Reference 11

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

January-February 1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

not specified

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes

Specific details on test material used for the study:

- Analytical purity: 99.8%
- Impurities: Na2O, Fe2O3, CaO
- Storage condition of test material: room temperature

Analytical monitoring:

yes

Details on sampling:

- Concentrations: test concentration and control
- Sampling method: concentration of aluminium in solution was recorded during the equilibrium after 1,2,3,4,7 days and at the start and end of the toxicity test
- Sample storage conditions before analysis: samples were preserved by addition of 1 mL 6M HNO3 per 100 mL sample

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: 100 mg aluminium hydroxide was added to 1000 mL of dilution water in a 2 L Erlenmeyer flask which was placed in the dark on a reciprocal shaker.
- Eluate: after 9 days, the soluble fraction was siphoned out and used in the toxicity test and the pH was adjusted to 7.8
- All other template details: Not reported

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Strain: Strain A (Baird et al. 1992)
- Source: University of Cardiff, UK
- Stock culture: cultured in Elendt M7 medium with continuous feeding with green algae (Selenastrum capricornutum)
- Age at study initiation (mean and range, SD): < 24 hours old
- Feeding during test: none
- All other template details: Not reported

ACCLIMATION: Not reported

QUARANTINE (wild caught): Not applicable

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

Not reported

Hardness:

Not reported

Test temperature:

19.1 - 21.1 degrees C

pH:

7.8 ± 0.2

Dissolved oxygen:

8.4-8.5 mg/L

Salinity:

Not applicable

Nominal and measured concentrations:

Nominal (mg/L): 100

Details on test conditions:

TEST SYSTEM
- Test vessel: 50 mL polystyrene beaker
- Material, size, headspace, fill volume: 50 mL polystyrene beaker filled with 25-35 mL of solution (5 mL/animal)
- Aeration: none
- No. of organisms per vessel: 5-7
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- All other template details: Not reported

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dilution water was modified Elendt M7 medium
- See Table 1 for detailed composition of the test medium
- Culture medium different from test medium: no
- Intervals of water quality measurement: after 48 hours

OTHER TEST CONDITIONS
- Adjustment of pH: adjusted to pH 7.8 ± 0.2 by addition of HCl or NaOH
- Photoperiod: 24 hr dark
- Light intensity: none

EFFECT PARAMETERS MEASURED: immobilisation observed at 24 and 48 hours

TEST CONCENTRATIONS
- Spacing factor for test concentrations: only one test concentration
- All other template details: Not reported

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

> 5 µg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

dissolved

Basis for effect:

mortality

Remarks on result:

other: nominal 100 mg/L

Details on results:

- Mortality of control: none
- All other template details: Not reported

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- Mortality: yes
- EC50/LC50: 1.4 mg/L

Reported statistics and error estimates:

There were no mortalities at the highest concentration tested, therefore no statistics were needed.

Not applicable

Conclusions:

There were no mortalities observed with the nominal loading of 100 mg/L observed after 48 hours of exposure by Daphnia magna. The mean measured dissolved Al concentration was 5 µg/L.

Reference 12

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

January-February 1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

not specified

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes

Specific details on test material used for the study:

- Molecular formula: Al2O3
- Analytical purity: 99.3%
- Impurities: Fe2O3, Na2O, CaO
- Storage condition of test material: room temperature

Analytical monitoring:

yes

Details on sampling:

- Concentrations: control and test solution
- Sampling method: sampled during the eluation after 1,2,3,4,7 days and at the start of the toxicity test
- Sample storage conditions before analysis: addition of 1 mL of 6 M HNO3 per 100 ml sample

Vehicle:

not specified

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: 100 mg aluminium oxide was added to 1000 mL of dilution water in a 2 L Erlenmeyer flask which was placed in the dark on a reciprocal shaker.
- Eluate: after 9 days, the soluble fraction was siphoned out for the toxicity test
- All other template details: Not reported

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: fresh water crustacean, daphnid
- Strain: Strain A (Baird et al. 1992)
- Source: University of Cardiff, UK
- Stock culture: cultured in Elendt M7 medium with continuous feeding with green algae (Selenastrum capricornutum)
- Age at study initiation (mean and range, SD): < 24 hours old
- Feeding during test: none
- All other template details: Not reported

ACCLIMATION: Not reported

QUARANTINE (wild caught): Not applicable

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

Not reported

Hardness:

Not reported

Test temperature:

19.1-21.1 degrees C

pH:

7.8 ± 0.2

Dissolved oxygen:

8.4 mg/L

Salinity:

Not applicable

Nominal and measured concentrations:

Nominal (mg/L): 100

Details on test conditions:

TEST SYSTEM
- Test vessel: 50 mL polystyrene beaker
- Material, size, headspace, fill volume: 50 mL polystyrene beaker filled with 25-35 mL of solution (5 mL/animal)
- Aeration: none
- No. of organisms per vessel: 5-7
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- All other template details: Not reported

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dilution water was modified Elendt M7 medium
- See Table 1 for detailed composition of the test medium
- Culture medium different from test medium: no
- Intervals of water quality measurement: after 48 hours

OTHER TEST CONDITIONS
- Adjustment of pH: adjusted to pH 7.8 ± 0.2 by addition of HCl or NaOH
- Photoperiod: 24 hr dark
- Light intensity: none

EFFECT PARAMETERS MEASURED: immobilisation observed at 24 and 48 hours

TEST CONCENTRATIONS
- Spacing factor for test concentrations: only one test concentration
- All other template details: Not reported

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

> 71 µg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

dissolved

Basis for effect:

mortality

Remarks on result:

other: nominal 100 mg/L Al

Details on results:

- Mortality of control: none
- All other template details: Not reported

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- Mortality: yes
- EC50/LC50: 1.4 mg/L

Reported statistics and error estimates:

There were no mortalities at the highest concentration tested, therefore no statistics were needed.

Not applicable

Conclusions:

There were no mortalities observed with the nominal loading of 100 mg/L observed after 48 hours of exposure by Daphnia magna. The mean measured dissolved Al concentration was 71 µg/L.

Reference 13

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

January-February 1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

not specified

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes

Specific details on test material used for the study:

- Name of test material: Aluminium (metallic)
- Physical state: powder
- Analytical purity: 99.7
- Impurities: Fe, Si
- Storage condition of test material: room temperature

Analytical monitoring:

yes

Details on sampling:

- Concentrations: test concentration and control
- Sampling method: concentration of aluminium in solution was recorded during the equilibrium after 1,2,3,4,7 days and at the start and end of the toxicity test
- Sample storage conditions before analysis: samples were preserved by addition of 1 mL 6M HNO3 per 100 mL sample

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: 100 mg aluminium hydroxide was added to 1000 mL of dilution water in a 2 L Erlenmeyer flask which was placed in the dark on a reciprocal shaker.
- Eluate: after 9 days, the soluble fraction was siphoned out and used in the toxicity test and the pH was adjusted to 7.8
- All other template details: Not reported

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Strain: Strain A (Baird et al. 1992)
- Source: University of Cardiff, UK
- Stock culture: cultured in Elendt M7 medium with continuous feeding with green algae (Selenastrum capricornutum)
- Age at study initiation (mean and range, SD): < 24 hours old
- Feeding during test: none
- All other template details: Not reported

ACCLIMATION: Not reported

QUARANTINE (wild caught): Not applicable

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

Not reported

Hardness:

Not reported

Test temperature:

19.1 - 21.1 degrees C

pH:

7.8 ± 0.2

Dissolved oxygen:

8.4 mg/L

Salinity:

Not applicable

Nominal and measured concentrations:

Nominal (mg/L): 100

Details on test conditions:

TEST SYSTEM
- Test vessel: 50 mL polystyrene beaker
- Material, size, headspace, fill volume: 50 mL polystyrene beaker filled with 25-35 mL of solution (5 mL/animal)
- Aeration: none
- No. of organisms per vessel: 5-7
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- All other template details: Not reported

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dilution water was modified Elendt M7 medium
- See Table 1 for detailed composition of the test medium
- Intervals of water quality measurement: after 48 hours

OTHER TEST CONDITIONS
- Adjustment of pH: adjusted to pH 7.8 ± 0.2 by addition of HCl or NaOH
- Photoperiod: 24 hr dark
- Light intensity: none

EFFECT PARAMETERS MEASURED: immobilisation observed at 24 and 48 hours

TEST CONCENTRATIONS
- Spacing factor for test concentrations: only one test concentration
- All other template details: Not reported

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

> 135 µg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

dissolved

Basis for effect:

mortality

Remarks on result:

other: nominal 100 mg/L

Details on results:

- Mortality of control: none
- All other template details: Not reported

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- Mortality: yes
- EC50/LC50: 1.4 mg/L

Reported statistics and error estimates:

No mortality observed, no statistics required

Not applicable

Conclusions:

There were no mortalities observed with the nominal loading of 100 mg/L observed after 96 hours of exposure by Daphnia magna. The mean measured dissolved Al concentration was 135 µg/L.

Reference 14

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Principles of method if other than guideline:

test method is described in the publication

GLP compliance:

not specified

Specific details on test material used for the study:

Al2SO4

Details on sampling:

A 20 mL water sample was collected from each treatment tank for analytical testing.T
o determine dissolved Al content, samples were passed through a 0.45 μm cellulose acetate filter and then acidified, with the total recoverable Al obtained through a 1-h hotblock digest using 10 mL of 70% nitric acid. Samples were made up to a final volume of 25 mL with high purity Milli-Q and then diluted 10x for analysis.

Details on test solutions:

A 4.6 g/L Al stock solution was prepared by dissolving Al sulphate (Al2(SO4), Chem-Supply, Gillman, South Australia) into estuary water (pH 8.23 ± 0.04 [mean ± SE]; salinity 35.4 ± 0.3 [mean ± SE]).

Test organisms (species):

other aquatic arthropod: Metapenaeus macleayi

Details on test organisms:

Juvenile School Prawn (carapace length [CL] 5–10 mm) were collected from the Hunter River estuary (on the mid-north coast of New South Wales, Australia).
Prawns were sorted and identified immediately after retrieval of the net. Prawns were then im- mediately placed in an onboard aerated holding tank, and transported to the Port Stephens Fisheries Institute (PSFI) Research Aquarium. At PSFI, animals were acclimated to environmental conditions at the aquarium over 48 h (32 salin- ity, 22 °C, pH 8.2) and held in a 5000 L aerated polyethylene tank with a 25% exchange of water each day. Animals were held under these conditions for 8 days prior to testing, and fed ad libitum with a dry feed (Otohime Hirame® by Aquasonic) in the evenings (approximately 5–10 g).

Test type:

semi-static

Water media type:

saltwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

22°C

pH:

8 and 5
Al the pH of the water was adjusted in small increments using strong sulphuric acid or 8 M sodium hydroxide/estuary water solutions.

Nominal and measured concentrations:

nominal: 2, 4, 8, 16, 32, 64, 128 mg Al/L

Details on test conditions:

Experimental treatments were undertaken in 6 L aquaria tanks in a light (12-h light/dark) controlled room, and 3 tanks per concentration.
School Prawn used in the experiments were 5.99 ± 0.03 mm CL (mean ± SE) and 0.28 ± 0.03 g (w/w).
Prawns were unfed for 24 h prior to testing, and unfed during the experimental period.

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 128 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

element

Basis for effect:

mortality

Details on results:

There was minimal mortality observed following exposure to Al across the ranges tested, with the majority of pH 8 treatments showing 90% survival or greater (98.9 ± 0.6%; mean ± SE, averaged across all concentrations). Survival was lower in the pH 5 treatment (75.2 ± 4.1%; mean ± SE, averaged across all concentrations), but did not appear to form a consistent relationship with the concentration of total Al. These relationships were confirmed by the statistical analysis, where pH had a significant effect on survival (F1,50= 31.92, P < < 0.01), but there was no significant effect of total Al (F1,50= 0.49,P= 0.49), or interaction between Al and pH (F1,50= 0.59,P= 0.45). Similar patterns were obtained when examining survival with dissolved Al, where lower survival was observed at pH 5 (F1,50= 31.27, P < < 0.01), but with no significant effect of dissolved Al (F1,50= 0.04,P= 0.84) nor significant interaction (F1,50= 0.01,P= 0.95).

Reported statistics and error estimates:

An analysis-of-covariance was used to analyse the interactive effect of pH (fixed; 2-levels; pH 5 and pH 8), and log10(total Al), on log10(dissolved Al), survival and bioaccumulated tissue Al concentration (logarithmic transformations were performed to linearise the data). Dispersion of survival data between pH 8 and pH 5 treatments was evaluated using the betadisper function in R package vegan (Oksanen et al. 2017). Further ANCOVA’s were conducted to assess the interactive effect of pH and log10(- dissolved Al), on survival and log10(tissue Al). All analyses were undertaken using R v.3.2.0 (R Core Team 2016).

Aluminium was more soluble at lower exposure concentrations but began to form visible precipitate as Al exposure concentra- tions increased (>~100.5mg L−1). Due to this variability in solubility, actual exposure concentrations were slightly lower than target concentrations.

Conclusions:

Exposure to low pH and Al contributed to mortality in School Prawn, and led to increased bioaccumulation of Al, structural degradation, and infection. Toxicity of Al may be amplified where other stressors are involved, but this requires further investigation. Furthermore, it would be useful to derive EC50 or EC10 for other more sensitive sub-lethal endpoints such as swimming speed, balance, and growth, to further explore sub-lethal impacts on health at lower exposure concentrations.
The lethal and sub-lethal effects of the stressors examined here could impact productivity of School Prawn if similar conditions occur in estuarine systems. Low pH conditions (pH 5), and high dissolved Al concentrations definitely occur in south-eastern Australian estuarine systems. This may have flow-on effects for fisheries productivity in this and other valuable crustacean species in south-eastern Australia such as Blue Swimmer Crab (Portunus armatus) and Giant Mud Crab (Scylla serrata). Such impacts create an impetus for catchment repair especially where acid sulphate soils are exposed and subject to oxidation, which is a significant problem for subtropical coastal catchments in south-eastern Australia. Catchment and habitat repair may alleviate productivity bottlenecks resulting from the water quality variables examined here, and thus represents an opportunity to improve the productivity and profitability of coastal fisheries.

Executive summary:

Catchment degradation and exposure of acid sulphate soils can affect estuarine water quality, and this can have impacts on the health of estuarine species and adversely affect fishery productivity. In degraded catchments, aluminium (Al) is mobilised from clay minerals following oxidation of acid sulphate soils, and may be harmful to estuarine crustaceans. We tested the acute toxicity and sub-lethal effects of Al for School Prawn (Metapenaeus macleayi), through a series of experiments conducted under normal (pH 8) and acidic (pH 5) conditions. Experimental data were used to examine mortality. Also, histological examination of the gills and hepatopancreas was conducted to determine pathological consequences of exposure to these stressors. School Prawn did not experience mortality in response to acute exposure to Al under normal pH conditions, but mortality and tissue bioaccumulation of Al was greater under acidic conditions, suggesting an interactive effect of both stressors. Histology revealed sub-lethal effects of Al including structural abnormal- ities in the gills and hepatopancreas, and evidence of viral infection and immune response, particularly at lower pH and higher Al concentrations. These impacts may impede major vital functions such as respiration, osmotic regulation, metabolism and growth of juvenile School Prawn, which could contribute to productivity bottlenecks in degraded estuaries.

Reference 15

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

significant methodological deficiencies

GLP compliance:

not specified

Specific details on test material used for the study:

AAS standards of aluminium, Sigma-Aldrich Co.

Test organisms (species):

other: Asplanchna brightwellii

Details on test organisms:

Asplanchna brightwellii females carrying embryos were placed in a 24-well polystyrene plate (one female per well to avoid canni- balism), then incubated at 25 °C, photoperiod 16:8 (l:d) in absence of food. Neonates less than 24-hr-old were collected next day and used in testing.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

24 h

Nominal and measured concentrations:

Nominal: 0.1, 0.5, 1, 1.5, 2.0 mg/L

Details on test conditions:

Ten neonate females were placed in bottles of 20 ml total volume; with a test volume of 5 ml.
ach test consisted of five replicates, negative control EPA medium pH 7.5 (USEPA, 1985), and five metal concentrations (atomic absorption standards of aluminum).
The animals were Incubated for 24 hours at 25 °C, photoperiod 16:8 (l:d) in absence of food.

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

0.174 mg/L

Nominal / measured:

not specified

Conc. based on:

element

Basis for effect:

mortality

Remarks on result:

other: CL/ 0.089-0.338 mg Al/L

Reported statistics and error estimates:

A one-way analysis or variance (ANOVA) and Duncan’s test were calculated to compare mortality percentages for each toxicant concentration to that of the control. The LC50 values were calculated using regression between probit units and the logarithm of each toxicant concentration using the software Sta- tistica 6.0 (StatSoft Inc., 2001).

Conclusions:

Tests conducted with AAS standards have shown to result in very low effect levels, often due to the very low pH of the standard solution that is used as test material. As no indication is given thar pH was measured or corrected, test data are not considered reliable.

Executive summary:

Acute toxicity tests of five metals (aluminum, cadmium, iron, lead, zinc) were performed to determine LC50 values in two species of freshwater rotifers: Asplanchna brigthwellii and its prey Brachionus calyciflorus. We conducted the tests using neonates less than 24 hr-old, each test consisted of five replicates, negative control and five metal concentrations (Al, Cd, Fe, Pb, Zn). We found that the prey rotifer B. calyciflorus was more sensitive to Al, Cd, Pb and Fe than the predator rotifer A. brightwellii. For both rotifers Cd was the most toxic of the five metals. It was established that the strain of B. ca- lyciflorus studied is sensitive when compared with other B. calyciflorus strains and other species and genera of the family Brachionidae. In the other hand, LC50 values of A. brigthwellii are compared with rotifer and copepod predators.

Reference 16

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

significant methodological deficiencies

GLP compliance:

not specified

Specific details on test material used for the study:

AAS standards of aluminium, Sigma-Aldrich Co.

Test organisms (species):

other: Brachionus calyciflorus

Details on test organisms:

Brachionus calyciflorus neonates less than 24-hr-old hatching from cysts were placed in a 24-well polystyrene plate (ten neonates per well) with a test volume of 1 ml.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

24 h

Nominal and measured concentrations:

Nominal: 0.01, 0.05, 0.25, 1.0, 1.75 mg/L

Details on test conditions:

Ten neonate females were placed in bottles of 20 ml total volume; with a test volume of 5 ml.
ach test consisted of five replicates, negative control EPA medium pH 7.5 (USEPA, 1985), and five metal concentrations (atomic absorption standards of aluminum).
The animals were Incubated for 24 hours at 25 °C, photoperiod 16:8 (l:d) in absence of food.

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

0.105 mg/L

Nominal / measured:

not specified

Conc. based on:

element

Basis for effect:

mortality

Remarks on result:

other: CL: 0.056 - 0.197 mg Al/L

Reported statistics and error estimates:

A one-way analysis or variance (ANOVA) and Duncan’s test were calculated to compare mortality percentages for each toxicant concentration to that of the control. The LC50 values were calculated using regression between probit units and the logarithm of each toxicant concentration using the software Sta- tistica 6.0 (StatSoft Inc., 2001).

Conclusions:

Tests conducted with AAS standards have shown to result in very low effect levels, often due to the very low pH of the standard solution that is used as test material. As no indication is given thar pH was measured or corrected, test data are not considered reliable.

Executive summary:

Acute toxicity tests of five metals (aluminum, cadmium, iron, lead, zinc) were performed to determine LC50 values in two species of freshwater rotifers: Asplanchna brigthwellii and its prey Brachionus calyciflorus. We conducted the tests using neonates less than 24 hr-old, each test consisted of five replicates, negative control and five metal concentrations (Al, Cd, Fe, Pb, Zn). We found that the prey rotifer B. calyciflorus was more sensitive to Al, Cd, Pb and Fe than the predator rotifer A. brightwellii. For both rotifers Cd was the most toxic of the five metals. It was established that the strain of B. ca- lyciflorus studied is sensitive when compared with other B. calyciflorus strains and other species and genera of the family Brachionidae. In the other hand, LC50 values of A. brigthwellii are compared with rotifer and copepod predators.

Reference 17

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Principles of method if other than guideline:

method described in the publication

GLP compliance:

not specified

Specific details on test material used for the study:

analytical grade metallic salt: Al2(SO4)3.18H2O

Analytical monitoring:

yes

Details on sampling:

Samples of water for metal analysis taken before and immediately after each solution renewal were acidified to 1% with ARISTAR nitric acid (65%) (BDH Inc, VWR International Ltd., England) before metal analysis

Details on test solutions:

The stock solutions were prepared with deionized water in 1L volumetric flasks. Metal solutions were prepared by dilution of a stock solution with dechlorinated tap water. A control with dechlorinated tap water only was also used.

Test organisms (species):

other aquatic mollusc: Melanoides tuberculata

Details on test organisms:

Snails M. tuberculata were collected from canals in the university in Bangi, Selangor, Malaysia. Prior to toxicity testing, the snails were acclimatized for one week under laboratory conditions (28–30°C) with 12:12 L:D in 50L stocking tanks using dechlorinated tap water (filtered by several layers of sand and activated carbon; T.C. Sediment Filter (TK Multitrade, Seri Kem- bangan, Malaysia)) aerated through an air stone. During acclimation the snails were fed on lettuce.
Test organisms were adult snails (shell length approx. 1.5-2.0 cm, mean wet weight 22.5 +/- 1.6 mg)

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

28-30°C

Nominal and measured concentrations:

Nominal: 56, 100, 320, 560, 1000 mg/K
Measured: 88.38, 160,83, 362.83, 884.34, 1229.91 mg Al/L

Details on test conditions:

The tests were carried out under static conditions with renewal of the solution every two days.
Control and metal-treated groups each consisted of two replicates of five randomly allocated snails in a 500mL glass beaker containing 400mL of the appropriate solution.
A total of 10 animals per treatment/concentration were used in the experiment
No stress was observed for the snails in the solution, indicated by 100% survival for the snails in the control water until the end of the study.
During the toxicity test, the snails were not fed.
Photoperiod 12 h light : 12 h darkness, using fluorescent lights (334–376 lux).

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

880.78 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

element

Basis for effect:

mortality

Remarks on result:

other: CL: 553.9-2147.6 mg Al/L

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

306.89 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

element

Basis for effect:

mortality

Remarks on result:

other: CL: 184.3-487.2 mg Al/L

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

130.22 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

element

Basis for effect:

mortality

Remarks on result:

other: CL: 35.51-226.38 mg Al/L

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

68.23 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

element

Basis for effect:

mortality

Remarks on result:

other: CL: 2.24-123.87 mg Al/L

Reported statistics and error estimates:

Median lethal concentrations (LC50) for the snails exposed to metals were calculated using measured metal concentrations. FORTRAN programs based on the methods of Litchfield [44] and Litchfield and Wilcoxon [45] were used to compute the LC50. Data were analyzed using concentration/response (CR) methods by plotting cumulative percentage mortality on logarithmic-probit paper.

Executive summary:

Adult freshwater snails Melanoides tuberculata (Gastropod, Thiaridae) were exposed for a four-day period in laboratory conditions to a range of copper (Cu), cadmium (Cd), zinc (Zn), lead (Pb), nickel (Ni), iron (Fe), aluminium (Al), and manganese (Mn) concentrations. Mortality was assessed and median lethal times (LT50) and concentrations (LC50) were calculated. LT50 and LC50 increased with the decrease in mean exposure concentrations and times, respectively, for all metals. The LC50 values for the 96- hour exposures to Cu, Cd, Zn, Pb, Ni, Fe, Al, and Mn were 0.14, 1.49, 3.90, 6.82, 8.46, 8.49, 68.23, and 45.59 mg L−1 , respectively. Cu was the most toxic metal to M. tuberculata, followed by Cd, Zn, Pb, Ni, Fe, Mn, and Al (Cu > Cd > Zn > Pb > Ni > Fe > Mn > Al). Metals bioconcentration in M. tuberculata increases with exposure to increasing concentrations and Cu has the highest accumulation (concentration factor) in the soft tissues. A comparison of LC50 values for metals for this species with those for other freshwater gastropods reveals that M. tuberculata is equally sensitive to metals.

Reference 18

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

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

Remarks:

Summary of available data used for the endpoint assessment of the target substance

Adequacy of study:

weight of evidence

Justification for type of information:

Refer to analogue justification provided in IUCLID section 13.

Reason / purpose for cross-reference:

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Duration:

48 h

Dose descriptor:

other: EC50/LC50

Effect conc.:

>= 0.071 - <= 99.6 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

element

Basis for effect:

mobility

Remarks on result:

other: range is from 0.071 - >99.6 mg Al/L, not normalised

Remarks:

Ceriodaphnia sp. and Daphnia sp.

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 22 - <= 30.6 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

element

Basis for effect:

mobility

Remarks on result:

other: Results from not normalised data; Results for Acronuria sp. only show 96-h NOEC of >22.6 mg Al/L, thus LC50 must be >22.6 mg/L Al

Remarks:

non-standard species Acronuria sp., Gammarus sp. and Physa sp.

Description of key information

Reliable (K1, K2) acute toxicity data were identified for five invertebrate species, and the majority of reported effect levels were obtained with the standard species D. magna and C. dubia. The available 48-h EC/LC50 values varied from 0.071 to > 99.6 mg Al/L. The acute NOECs (48 h) varied from > 0.005 to > 0.135 mg Al/L. Most of the variation in results can be explained by differences in hardness and DOC in the test media.

Acute toxicity for the non-standard species Acronuria sp., Gammarus sp. And Physa sp. – expressed as mg Al/L – were >22.6, 22 and 30.6 mg/L, respectively.

Additional information

Effect levels were expressed as mg Al/L (total, dissolved), but the complex speciation properties of Al and its low solubility make the interpretation of reported effect levels challenging. This is reflected in the results that were generated by CIMM (2009) with C. dubia where the difference in EC50 expressed as either dissolved aluminum or total aluminium was up to three orders of magnitude.

For a detailed overview of the data, more information is provided in the Background document "Environmental Effects Assessment of Aluminium" attached to IUCLID section 13.