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Toxicity to aquatic algae and cyanobacteria

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Endpoint:
toxicity to aquatic algae and cyanobacteria
Type of information:
read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
2002-09-24 to 2002-09-27
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
Guideline and GLP study, no deviations. The reliability of this study for this substance tested is a K1, but in application of read-across to a different substance, ECHA's guidance specifies that the score can be a maximum of a K2.
Justification for type of information:
1. HYPOTHESIS FOR THE CATEGORY APPROACH: The hypothesis is that properties are likely to be similar or follow a similar pattern because of the presence of a common metal ion, in this case tungstate.
2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES):
Source: Ammonium paratungstate
Target: Ammonium metatungstate
3. CATEGORY APPROACH JUSTIFICATION: See Annex 3 in CSR
4. DATA MATRIX: See Annex 3 in CSR
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
according to guideline
Guideline:
OECD Guideline 201 (Alga, Growth Inhibition Test)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method C.3 (Algal Inhibition test)
Deviations:
no
Principles of method if other than guideline:
N/A
GLP compliance:
yes
Remarks:
stated, no certificate
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
N/A
Analytical monitoring:
yes
Details on sampling:
- Concentrations: All concentrations
- Sampling method: 0.5 mL hydrofluoric acid (w=40%) was immediately added to each sample to stabilize it. The samples were then assumed stable for up to 8 weeks.
- Sample storage conditions before analysis: Room temperature.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: a stock solution was prepared to give the desired series of test concentrations. 125 mg of test substance were added to 1 L of dilution water and treated for 1 hour in an ultrasonic bath. The pH was adjusted to 7.8 and the stock solution was stirred for 24 hours on a magnetic stirrer.
- Eluate: N/A
- Differential loading: N/A
- Controls: Algal medium only
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): N/A
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)):N/A
- Evidence of undissolved material (eg precipitate, surface film, etc: no
Test organisms (species):
Desmodesmus subspicatus (previous name: Scenedesmus subspicatus)
Details on test organisms:
TEST ORGANISM
- Common name: Green algae
- Strain: N/A
- Source (laboratory, culture collection): Non-axenic strain obtained from The Collection of Algal Cultures of the Institute of Plant Physiology, at the University of Gottingen, Germany
- Age of inoculum (at test initiation): 3 days
- Method of cultivation: Pre-cultures are set up 3 days before test initiation from exponentially growing stock cultures and maintained under constant temperature and light.

ACCLIMATION
- Acclimation period:3 days, acclimated from Bringmann and Kuhn medium to test medium during this period
- Culturing media and conditions (same as test or not):same media and conditions as test (see above).
- Any deformed or abnormal cells observed: none
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
72 h
Post exposure observation period:
N/A
Hardness:
N/A
Test temperature:
23+/- 2 °C
pH:
Initial-7.8
Final-range 10.1-10.5
Dissolved oxygen:
N/A
Salinity:
N/A
Nominal and measured concentrations:
Nominal- Control, 6.3, 13, 25, 50 and 100 mg/L
Measured- Control, 5.8, 11.521.4, 45.4 and 88.3 mg/L.(WO3). Measured concentrations ranged from 95-102.8% of nominal as W at 0 hours, and from 100-104.2% of nominal at 72 hours.
Details on test conditions:
TEST SYSTEM
- Test vessel: Erlenmeyer flask
- Type (delete if not applicable): closed with stopper
- Material, size, headspace, fill volume: Glass, 300 mL, 100 mL
- Aeration: No
- Type of flow-through (e.g. peristaltic or proportional diluter): N/A
- Renewal rate of test solution (frequency/flow rate): N/A
- Initial cells density: 10^4 cells/mL
- Control end cells density: 3.15 x 10^5 cells/mL
- No. of organisms per vessel: N/A
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 6
- No. of vessels per vehicle control (replicates): N/A

GROWTH MEDIUM
- Standard medium used: yes
- Detailed composition if non-standard medium was used: N/A

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:algal medium used
- Total organic carbon: N/A
- Particulate matter: N/A
- Metals: N/A
- Pesticides: N/A
- Chlorine: N/A
- Alkalinity: N/A
- Ca/mg ratio: N/A
- Conductivity: N/A
- Culture medium different from test medium: yes
- Intervals of water quality measurement: N/A

OTHER TEST CONDITIONS
- Sterile test conditions: not stated
- Adjustment of pH: yes
- Photoperiod: continuous light
- Light intensity and quality: 4000-8000 lux (60 -120 microEinsteins per m^2 per sec)
- Salinity (for marine algae): N/A

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : cell density
- Determination of cell concentrations: microcell counter
- Chlorophyll measurement: N/A
- Other: N/A

TEST CONCENTRATIONS
- Spacing factor for test concentrations: approximately 2
- Justification for using less concentrations than requested by guideline: N/A
- Range finding study N/A
- Test concentrations: N/A
- Results used to determine the conditions for the definitive study: N/A
Reference substance (positive control):
no
Duration:
72 h
Dose descriptor:
EC50
Effect conc.:
> 70.5 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
element
Remarks:
as measured W
Basis for effect:
biomass
Duration:
72 h
Dose descriptor:
EC50
Effect conc.:
> 70.5 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
element
Remarks:
as measured W
Basis for effect:
growth rate
Duration:
72 h
Dose descriptor:
NOEC
Effect conc.:
36.5 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
element
Remarks:
as measured W
Basis for effect:
biomass
Duration:
72 h
Dose descriptor:
LOEC
Effect conc.:
70.5 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
element
Remarks:
as measured W
Basis for effect:
biomass
Duration:
72 h
Dose descriptor:
NOEC
Effect conc.:
36.5 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
element
Remarks:
as measured W
Basis for effect:
growth rate
Duration:
72 h
Dose descriptor:
LOEC
Effect conc.:
70.5 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
element
Remarks:
as measured W
Basis for effect:
growth rate
Details on results:
- Exponential growth in the control (for algal test): yes
- Observation of abnormalities (for algal test): none
- Unusual cell shape: N/A
- Colour differences: N/A
- Flocculation: N/A
- Adherence to test vessels: N/A
- Aggregation of algal cells: N/A
- Other: N/A
- Any stimulation of growth found in any treatment: yes, stimulation was observed in all test concentrations except the highest. Stimulation of growth ranged from less than 1 percent to approximately 5 percent.
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: none
- Effect concentrations exceeding solubility of substance in test medium: none
Results with reference substance (positive control):
N/A
Reported statistics and error estimates:
Deteminations of NOECs and LOECs were made by Dunnett's multiple t-test.

Table 1. Results Summary

 Concentration (mg/L) Avg.biomass (area under growth curve)   %inhibition  Growth rate (per day)  % inhibition
 Control  378611 0.0   1.15  0.0
6.3  395000  -4.3  1.16  -1.1 
13  395556  -4.5  1.16  -0.8 
25  397778  -5.1  1.16  -1.2 
50  385556  -1.8  1.17  -1.5 
 100

 257778

 31.9

 1.09

 5.4

a negative % inhibition indicates stimulation

Validity criteria fulfilled:
yes
Remarks:
Control increased by > 16 times.
Conclusions:
The 72 hour EC50 of the test substance to D. subspicatus was found to be >70.5 mg/L based on growth rate. The NOEC is 36.5 mg/L, and the LOEC is 70.5 mg/L, based upon growth rate.
Executive summary:

No toxicity to aquatic algae and cyanobacteria data of sufficient quality were available specifically on ammonium metatungstate (target substance). However, aquatic algae and cyanobacteria toxicity data are available on ammonium paratungstate (source substance), which are used for read-across. Due to similar water solubility for the target substance compared to the source substance, the resulting read-across from the source substance to the target substance is appropriate to estimate of potential toxicity for this endpoint. For more details, refer to the read-across category approach description in the Category section of this IUCLID submission or Annex 3 of the CSR.

Endpoint:
toxicity to aquatic algae and cyanobacteria
Type of information:
read-across based on grouping of substances (category approach)
Adequacy of study:
supporting study
Study period:
2008-10-06 to: 2009-03-19
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Well documented, scientifically sound study conducted in accordance to GLP and OECD guideline 201 with no deviations to the protocol. The reliability of this study for the substance tested is a K1, but in application of read-across to a different substance, ECHA's guidance specifies that the score can be a maximum of a K2.
Justification for type of information:
1. HYPOTHESIS FOR THE CATEGORY APPROACH: The hypothesis is that properties are likely to be similar or follow a similar pattern because of the presence of a common metal ion, in this case tungstate.
2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES):
Source: Sodium Tungstate
Target: Ammonium metatungstate
3. CATEGORY APPROACH JUSTIFICATION: See Annex 3 in CSR
4. DATA MATRIX: See Annex 3 in CSR
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
according to guideline
Guideline:
OECD Guideline 201 (Alga, Growth Inhibition Test)
Deviations:
no
GLP compliance:
yes
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
no data
Analytical monitoring:
yes
Details on sampling:
- Concentrations: A volume of 10 mL was collected from each control and test substance treatment at 0 and 72 hours and transferred to a culture tube. The samples at 0 hour were collected from the parent solutions and the samples at 72 hours were collected after combining replicate solutions by treatment. An abiotic replicate of the lowest test substance treatment was also sampled at 72 hours.
- Sampling method: 10 mL taken from the parent solutions at time 0, and from replicate composites at 72 hours. Samples were centrifuged for 15minutes and a 0.05 mL volume of the supernatant was taken, diluted as necessary with 2 % HNO3, capped and shaken to mix, and analyzed by ICP-MS.
- Sample storage conditions before analysis: Room temperature
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: 0.0200 g sodium tungstate was dissolved in 1,000 mL of freshwater algal nutrient medium. This primary standard was used as the highest test substance treatment and the five lower test substance treatments, each at a volume of 500 mL, were prepared individually using appropriate volumes of the primary standard and test medium.
- Controls: Algal nutrient medium only.

- The medium was prepared by the addition of appropriate reagent grade salts to autoclaved ABC reagent water. ABC reagent water is produced by passing reverse-osmosis water through a series of deionization tanks, a laboratory water purification system consisting of carbon, de-mineralisation, and organic adsorption cartridges, and then through a 0.2-um filter. After preparation, the medium was pH-adjusted to 7.5 ± 0.1 using 0.1 N NaOH and filtered through a 0.45-um Millipore® filters.
Test organisms (species):
Pseudokirchneriella subcapitata (previous names: Raphidocelis subcapitata, Selenastrum capricornutum)
Details on test organisms:
TEST ORGANISM
- Common name: Pseudokirchneriella supcapitata (formerly known as Selenastrum caprictum)
- Source (laboratory, culture collection): Department of Botany, Culture Collection of Algae, University of Texas at Austin
- Age of inoculum (at test initiation): 3 days
- Method of cultivation: Cloned from existing culture derived from parent stock (identified as Selenastrum capricornatum)

ACCLIMATION
- Acclimation period: None necessary; culture media and conditions the same as test conditions.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
72 h
Post exposure observation period:
None
Hardness:
n/a
Test temperature:
22.9-24.0 °C
pH:
7.5-7.9 SU
Dissolved oxygen:
N/A
Salinity:
N/A
Nominal and measured concentrations:
Nominal- Control, 0.39, 0.85, 1.9, 4.1, 9.1, and 20 mg/L
Mean calculated concentrations- 0.350,0.812, 1.79, 3.83, 7.96, and 17.62 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: 250-mL Erlenmeyer flasks with foam stoppers and labeled with study number, treatment, replicate, and grid position. Each test substance treatment was replicated three times.
- Initial cells density: 5x10E3 cells/mL
- Control end cells density: 113x10E4; 224 times initial density
- No. of vessels per concentration (replicates): 3; 4 (which included the additional replication of the lowest test substance treatment.
- No. of vessels per control (replicates): 6

GROWTH MEDIUM
- Standard medium used: yes

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Autoclaved ABC reagent water; Deionized reverse osmosis water passed through carbon, de-mineralization, and organic absorbtion cartridges and then a 0.2 um filter.
- Culture medium different from test medium: No

OTHER TEST CONDITIONS
- Sterile test conditions: yes
- Adjustment of pH: Adjusted to 7.5 ± 0.1 with 0.1 N NaOH and filtered through 0.45 um filter
- Photoperiod: Continuous light
- Light intensity and quality: 7,923-8,290 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Growth rate and Yield, 24 hours
- Determination of cell concentrations: Direct microscopic counting with a hemacytometer

TEST CONCENTRATIONS
- Spacing factor for test concentrations: Approx 2.2
- Justification for using less concentrations than requested by guideline:
- Range finding study
- Test concentrations: Range-finding test #1: Nominal concentrations of 0 (control), 0.010, 0.10, 1.0, 10, and 100 mg/L
Range-finding test #2: Nominal concentrations of 0 (control), 0.10, 1.0, 5.0, 10, and 50 mg/L
- Results used to determine the conditions for the definitive study: Yes
Reference substance (positive control):
no
Duration:
72 h
Dose descriptor:
EC50
Effect conc.:
> 17.7 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Remarks:
sodium tungstate
Basis for effect:
growth rate
Remarks on result:
other: 95 % CL could not be calculated
Duration:
72 h
Dose descriptor:
EC20
Effect conc.:
13.8 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Remarks:
sodium tungstate
Basis for effect:
growth rate
Remarks on result:
other: 95 % CL 13.0-14.6 mg/L
Duration:
72 h
Dose descriptor:
EC10
Effect conc.:
5.76 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Remarks:
sodium tungstate
Basis for effect:
growth rate
Remarks on result:
other: 95 % CL 4.94-6.58 mg/L
Duration:
72 h
Dose descriptor:
NOEC
Effect conc.:
0.812 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Remarks:
sodium tungstate
Basis for effect:
growth rate
Duration:
72 h
Dose descriptor:
EC50
Effect conc.:
7.35 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Remarks:
sodium tungstate
Basis for effect:
biomass
Remarks on result:
other: 95 % CL 6.58-8.12 mg/L
Duration:
72 h
Dose descriptor:
EC20
Effect conc.:
2.02 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Remarks:
sodium tungstate
Basis for effect:
biomass
Remarks on result:
other: 95 % CL 1.54-2.50 mg/L
Duration:
72 h
Dose descriptor:
EC10
Effect conc.:
0.95 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Remarks:
sodium tungstate
Basis for effect:
biomass
Remarks on result:
other: 95 % CL 0.626-1.27 mg/L
Duration:
72 h
Dose descriptor:
NOEC
Effect conc.:
0.812 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Remarks:
sodium tungstate
Basis for effect:
biomass
Details on results:
The number of algal cells in the control at test termination was greater than 16 times the number initially inoculated to verify logarithmic phase growth. The coefficient of variation for daily growth rates in the control replicates during the course of the test did not exceed 35%. The coefficient of variation of average specific growth rates during the whole test period in control replicate did not exceed 7%. Since the study was not conducted as a limit test, there were at least one test concentration exhibiting less than 50% decrease in growth and once concentration exhibiting greater than 50% decrease in growth relative to the control. The pH in the control did not increase more than 1.5 units during the study.
Reported statistics and error estimates:
All statistical analyses were performed with SAS software. NOEC values were calculated using ANOVA and Dunnett's tests. Shapiro-Wilk's and Levene'stests were conducted to test for normality and homogeneity of variance. EC estimates were calculated using a logistic model. Parametric analyses were performed on the 72-hour growth rate and yield data. Non-parametric analyses were performed on the 24- and 48- hour growth rate and yield data.
Conclusions:
The 72 hour EC50 for growth rate for P.subcapitata was >17.7 mg sodium tungstate/L, while the EC10 for growth rate was 5.76 mg sodium tungstate/L, with 95 % confidence limits of 4.94 and 6.58 mg/L. The 72 hour EC50 for yield was 7.35 mg/L, with 95 % confidence limits of 6.58 and 8.12 mg/L.
Executive summary:

No aquatic algae and cyanobacteria toxicity to aquatic invertebrates data of sufficient quality were available specifically on ammonium metatungstate (target substance). However, aquatic algae and cyanobacteria toxicity to aquatic invertebrates data are available on sodium tungstate (source substance), which are used for read-across. Due to similar water solubility and toxicity for the target substance compared to the source substance, the resulting read-across from the source substance to the target substance is appropriate for this endpoint. In addition, read across is appropriate because the classification and labelling is similar for the source substance and target substance, the PBT/vPvB profile is the same, and the dose descriptors are, or are expected to be, conservative for the target substance. For more details, refer to the read-across category approach description in the Category section of this IUCLID submission or Annex 3 of the CSR.

Endpoint:
toxicity to aquatic algae and cyanobacteria
Type of information:
read-across based on grouping of substances (category approach)
Adequacy of study:
supporting study
Study period:
2009-07-24 to 2010-02-11
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
GLP and Guideline study without deviations. Well documented. The reliability of this study for this substance tested is a K1, but in application of read-across to a different substance, ECHA's guidance specifies that the score can be a maximum of a K2.
Justification for type of information:
1. HYPOTHESIS FOR THE CATEGORY APPROACH: The hypothesis is that properties are likely to be similar or follow a similar pattern because of the presence of a common metal ion, in this case tungstate.
2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES):
Source: Sodium Tungstate
Target: Ammonium metatungstate
3. CATEGORY APPROACH JUSTIFICATION: See Annex 3 in CSR
4. DATA MATRIX: See Annex 3 in CSR
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
according to guideline
Guideline:
OECD Guideline 201 (Alga, Growth Inhibition Test)
Deviations:
no
Principles of method if other than guideline:
N/A
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
- Concentrations: All concentrations
- Sampling method: 10 ml was collected from each test solution and control at 0 and 72 hours
- Sample storage conditions before analysis: no data
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A primary stock solution was prepared in reagent water at a concentration of 2.45 mg sodium tunstate./ml. The solution was stored at room temperature when not in use.
- Controls: algal medium only
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): none
- Evidence of undissolved material (eg precipitate, surface film, etc): All sodium tungstate test solutions appeared clear and colorless, with no visible precipitates, surface films, or undissolved test substance at initiation and throughout the test.
Test organisms (species):
Pseudokirchneriella subcapitata (previous names: Raphidocelis subcapitata, Selenastrum capricornutum)
Details on test organisms:
TEST ORGANISM
- Common name: Pseudokirchneriella subcapitata
- Source (laboratory, culture collection): Original stock culture obtained from the Department of Botany, Culture Collection of Algae, University of Texas at Austin, Jan 13, 2009.
- Age of inoculum (at test initiation): 4 days
- Method of cultivation: The prepared cultures were maintained in a temperature-controlled environmental chamber under continuous light. Periodically, new cultures were cloned from an existing culture derived from the parent stock. All cultures were maintained under the same conditions as those used for testing.


ACCLIMATION
- Acclimation period: N/A
- Culturing media and conditions (same as test or not): same as test
- Any deformed or abnormal cells observed: no data
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
72 h
Post exposure observation period:
none
Hardness:
no data
Test temperature:
22.8-23.8 °C
pH:
7.4-8.4
Dissolved oxygen:
no data
Nominal and measured concentrations:
Nominal-Control, 13, 25, 50, 100, and 200 mg sodium tungstate/L
Initial measured- Control, 10.8, 21.4, 43.5, 87.9, and 173 mg sodium tungstate/L
Mean measured- Control, 10.9, 21.9, 41.6, 84.5 and 168 mg sodium tungstate/L
Details on test conditions:
TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: 250 ml Erlenmeyer flasks with foam stoppers
- Aeration: no
- Initial cells density: 5000 cells/ml
- Control end cells density: 750000 cells/ml
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 6
- No. of vessels per vehicle control (replicates): N/A


GROWTH MEDIUM
- Standard medium used: yes
- Detailed composition if non-standard medium was used: standard


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: ABC well water passed through RO and then deionization tanks, a laboratory water purification system consisting of carbon, demineralization and organic absorption cartridges, and then passed through a 0.2 um filter. The water was autoclaved before use in preparation of algal medium. Characteristics below are for the ABC well water.
- Total organic carbon: no data
- Particulate matter: no data
- Metals: Ca- 83.1, Mg-33.1, K-7.93, Na-32.2 mg/L
- Pesticides: BDL
- Chlorine: no data
- Alkalinity: no data
- Ca/mg ratio: 1:2.5
- Conductivity: no data
- Culture medium different from test medium: no
- Intervals of water quality measurement: yearly, primary measurements taken at test initiation and test termination


OTHER TEST CONDITIONS
- Sterile test conditions: yes
- Adjustment of pH: yes, to 7.5 with 0.1N HCl
- Photoperiod: continuous light
- Light intensity and quality: cool white fluorescent, 7936-8479 lux
- Salinity (for marine algae): N/A


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : growth rate and cell density, 24 h
- Determination of cell concentrations: direct microscopic counting with a hemacytometer
- Chlorophyll measurement: no
- Other: none


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Justification for using less concentrations than requested by guideline: N/A
- Range finding study: Not performed
Reference substance (positive control):
no
Duration:
72 h
Dose descriptor:
EC50
Effect conc.:
54.1 mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Remarks:
calculated sodium tungstate concentrations
Basis for effect:
growth rate
Remarks on result:
other: 95% CL 41.1 to 67.1
Duration:
72 h
Dose descriptor:
EC50
Effect conc.:
52.9 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
other: calculated sodium tungstate concentrations
Basis for effect:
growth rate
Remarks on result:
other: 95% CL 40.5-65.3 mg/L
Duration:
72 h
Dose descriptor:
EC10
Effect conc.:
< 10.9 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
other: calculated sodium tungstate concentrations
Basis for effect:
growth rate
Duration:
72 h
Dose descriptor:
EC20
Effect conc.:
< 10.9 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
other: calculated sodium tungstate concentrations
Basis for effect:
growth rate
Duration:
72 h
Dose descriptor:
NOEC
Effect conc.:
< 10.9 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
other: calculated sodium tungstate concentrations
Basis for effect:
growth rate
Details on results:
- Exponential growth in the control (for algal test): yes
- Any stimulation of growth found in any treatment: none
- Any observations (eg precipitation) that might cause a difference between measured and nominal values: none
- Effect concentrations exceeding solubility of substance in test medium: none
Results with reference substance (positive control):
N/A
Reported statistics and error estimates:
NOEC values were estimated using one-way ANOVA procedure and two-tailed Dunnett's test. Shiro-Wilke's and Levene's tests were also used as appropriate. EC values were calculated using a logistic model. All statistical analysis was conducted with SAS software.
Validity criteria fulfilled:
yes
Conclusions:
The 72 hour ErC50 of the test substance to P.subcapitata was found to be 52.9 mg sodium tungstate/L, with 95% confidence limits of 40.5-65.3 mg sodium tungstate/L. The 72 hour NOErC was <10.9 mg sodium tungstate/L.
Executive summary:

No aquatic algae and cyanobacteria toxicity to aquatic invertebrates data of sufficient quality were available specifically on ammonium metatungstate (target substance). However, aquatic algae and cyanobacteria toxicity to aquatic invertebrates data are available on sodium tungstate (source substance), which are used for read across. Due to similar water solubility and toxicity for the target substance compared to the source substance, the resulting read-across from the source substance to the target substance is appropriate for this endpoint. In addition, read-across is appropriate because the classification and labelling is similar for the source substance and target substance, the PBT/vPvB profile is the same, and the dose descriptors are, or are expected to be, conservative for the target substance. For more details, refer to the read-across category approach description in the Category section of this IUCLID submission or Annex 3 of the CSR.

Description of key information

The toxicity of ammonium metatungstate to aquatic algae / cyanobacteria is estimated based on read-across from ammonium paratungstate (APT). In a 72-h test on Desmodesmus subspicatus (green algae) with APT, 72-h EC50s of >70.5 mg W/L were reported for both growth rate reduction and biomass inhibition. NOECs for these endpoints were determined by Dunnett’s multiple t-test to be 36.5 mg W/L. All effect levels were based on arithmetic mean measured concentrations. Two algal toxicity assays are available for sodium tungstate, two of which tested Pseudokirchneriella subcapitata (green algae). In the first assay, 72-h ErC50 and ErC10 values of 52.9 mg sodium tungstate/L (ca. 40 mg W/L) and <10.9 mg sodium tungstate/L (ca. <6.39 mg W/L) are reported, respectively. In the second assay with P. subcapitata, the 72-h ErC50, ErC10, and NOEC values (based on growth rate) are >17.7, 5.76 and 0.812 mg sodium tungstate/L (ca. >10.4, 3.38 and 0.476 mg W/L), respectively, based on mean measured concentrations.

Key value for chemical safety assessment

EC50 for freshwater algae:
40 mg/L
EC10 or NOEC for freshwater algae:
36.5 mg/L

Additional information

No algal toxicity data of sufficient quality are available for ammonium metatungstate (target substance). However, algal toxicity data are available for ammonium paratungstate and sodium tungstate (source substances), which were evaluated for read-across.

 

Due to similar or lower transformation/dissolution results for the target substance than the source substances, the resulting toxicity potential is expected to be similar or lower, so read-across is appropriate. In addition, read-across is justified because the classification and labelling is the same or less severe for the target substance and the PBT/vPvB profile is the same. Finally, the dose descriptors are, or are expected to be, sufficiently similar or higher for the target substance, and read-across to the source chemicals are adequately protective. For more details, refer to the read-across category approach description in the Category section of this IUCLID submission or Annex 3 of the CSR.