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Ecotoxicological information

Toxicity to aquatic algae and cyanobacteria

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Administrative data

Link to relevant study record(s)

Reference
Endpoint:
toxicity to aquatic algae and cyanobacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
24 March 1997 to 27 March 1997
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study (OECD and EU), according to GLP.
Qualifier:
according to guideline
Guideline:
OECD Guideline 201 (Alga, Growth Inhibition Test)
Deviations:
yes
Remarks:
Minor deviations that did not affect the validity of the study: pH increased by more than 1.5 units in some of the control and test vessels. Only 2 instead of 3 medium control flasks were included.
Qualifier:
according to guideline
Guideline:
EU Method C.3 (Algal Inhibition test)
Deviations:
yes
Remarks:
see above
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
Samples of the methanol stock solutions were analysed after preparation. It was not possible to determine the concentration of the test substance in the test media.
Vehicle:
yes
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: stock solution was prepared by direct addition of test substance to methanol
- Controls: one containing methanol at the same concentration as the test medium and one containing dilution water only
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): methanol
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): 0.01 %
- Evidence of undissolved material (e.g. precipitate, surface film, etc): no, during the test, all test media were clear, colourless solutions
Test organisms (species):
Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)
Details on test organisms:
TEST ORGANISM
- Strain: CCAP 278/4
- Source (laboratory, culture collection): axenic culture derived from the Culture collection of Algae and Protozoa, Institute of Freshwater Ecology, Ambleside, Cumbria, UK
- Age of inoculum (at test initiation): the quantity of inoculum introduced into each test vessel was sufficient to give a concentration of 10000 cells/mL.
- Method of cultivation: liquid culture

ACCLIMATION
- Acclimation period: no
- Culturing media and conditions (same as test or not): no data
- Any deformed or abnormal cells observed: no data
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
72 h
Remarks on exposure duration:
combined limit-range finding test
Post exposure observation period:
not applicable
Hardness:
no data
Test temperature:
24.8-25.5 °C
pH:
after 0 h: 7.7 in control, solvent control and test medium
after 72 h: 9.3, 9.5, 9.8 in control, solvent control and test medium, respectively
Dissolved oxygen:
no data
Salinity:
not applicable
Nominal and measured concentrations:
nominal concentration of methanol stock solutions: 2 mg/mL and 20 mg/mL
nominal concentration of test medium: 0.2 mg/L
measured concentrations of methanol stock solutions: 1.882 mg/mL and 18.552 mg/mL (94 % and 93 % of nominal concentration, respectively)
Concentrations of test substance could not be determined in test solutions, but only in stock solutions, immediately after preparation.
Details on test conditions:
TEST SYSTEM
- Test vessel: Erlenmeyer flask
- Type (delete if not applicable): closed: loose capped
- Material, size, headspace, fill volume: glass, 250-mL size, 100 mL fill volume of test medium
- Aeration: no
- Type of flow-through (e.g. peristaltic or proportional diluter): not applicable
- Renewal rate of test solution (frequency/flow rate): not applicable
- Initial cells density: nominal cell concentration: 10^4 cells/mL
- Control end cells density: 161-190 x 10^4 cells/mL
- No. of vessels per concentration (replicates): 5, of which 3 contained algae
- No. of vessels per control (replicates): 4, of which 2 contained algae
- No. of vessels per vehicle control (replicates): 8, of which 6 contained algae
Remaining vessels were used for background count and water quality.

GROWTH MEDIUM
- Standard medium used: yes
- Detailed composition if non-standard medium was used: OECD201-medium
- Medium was autoclaved before addition of NaHCO3.
- NaHCO3 was added to algal culture medium after filter sterilisation (0.2 µm filter).

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: deionised water
- Culture medium different from test medium: no
- Intervals of water quality measurement:at start and end of test

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: continuous illumination
- Light intensity and quality: 8050 lux at start of test, 9200 lux at end of test
- Incubation: cooled orbital incubator: 100 cycles/min

MONITORING:
- pH: determined at start and end of test in control and test media
- temperature: taken at start and at 24 h intervals until end of test in one control vessel
- light intensity: determined at start and end of test
- cell counts: performed at approximately 24 h intervals after start of incubation for all flasks (except flasks used for determination of water quatity)

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: electronic particle counter: Z1 coulter counter (Coulter Electronics Ltd, Luton, UK)
- Chlorophyll measurement: no
- Correction for cell counts were made by subtraction of background counts from a vessel incubated with the test vessels but containing no algae.
- Acceptability of electronic counting was carried out by comparison of cell concentrations using a haemocytometer and light microscope.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: not applicable
- Justification for using less concentrations than requested by guideline: a combined limit and range-finding test was performed and resulted in no growth inhibition to S.capricornutum after 72h. Therefore, no definitive test was performed and only results of limit test are reported.
- Range finding study: results not reported
Reference substance (positive control):
no
Duration:
72 h
Dose descriptor:
other: EbC50
Effect conc.:
> 0.2 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Remarks:
: area under growth curve
Duration:
72 h
Dose descriptor:
other: ErC50
Effect conc.:
> 0.2 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Remarks:
: average specific growth rate
Duration:
72 h
Dose descriptor:
NOEC
Effect conc.:
> 0.2 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Remarks:
: based on observation
Details on results:
The pH of the test media increased by more than the 1.5 units in some of the control and test vessels. Growth of the control cultures was greater than a factor 16 over the 72h test period demonstrating that the environmental conditions were acceptable for the test.
Results with reference substance (positive control):
not applicable
Reported statistics and error estimates:
Algal concentration data were evaluated using two approaches:
* comparison of areas under growth curve: EbC50
* comparison of average specific growth rate: ErC50
* NOEC was determined from results by observation (no statistical method could be applied to test the data because only one test concentration was used)

Results of the study are based on nominal concentrations because determination of test substance in test media was not possible.

Validity criteria fulfilled:
yes
Remarks:
pH increased with more than 1.5 in some control and test vessels but this did not negatively influence test results.
Conclusions:
Reliable study (Klimisch 1) reporting EbC50 / ErC50 > 0.2 mg/L and NOEC > 0.2 mg/L for Selenastrum capricornutum.

Description of key information

One GLP study was presented.

The Covance study conducted a guideline study (OECD 201/EC C.3) by exposing Selenastrum capricornutum for 72h in a static limit test to a nominal concentration of 0.2 mg/L of Wingstay L-HLS. The EC50 was >0.2 mg/L and the NOEC was 0.2 mg/L, based on nominal concentrations.

Key value for chemical safety assessment

EC50 for freshwater algae:
0.2 mg/L
EC10 or NOEC for freshwater algae:
0.2 mg/L

Additional information

Due to the low water solubility, the test substance in the Covance study was first dissolved in methanol (20 and 2 g/L stock solutions) and then used in the test medium to obtain a nominal concentration of 0.2 mg/L. No undissolved material was present. The concentrations in the stock solutions were measured and were 93 -94% of nominal.

No adverse growth effects occurred and no other effects were observed. It can be concluded that the substance does not cause any adverse effects to algae growth at concentrations which are higher than its water solubility.