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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:
25 August 2011 to 26 March 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reason / purpose for cross-reference:
reference to other study
Qualifier:
according to guideline
Guideline:
OECD Guideline 201 (Alga, Growth Inhibition Test)
Qualifier:
according to guideline
Guideline:
EU Method C.3 (Algal Inhibition test)
GLP compliance:
yes (incl. QA statement)
Analytical monitoring:
no
Details on sampling:
Not applicable. Due to low aqueous solubility of the test substance, it was not possible to develop a suitable method of analysis with sufficient sensitivity to verify concentrations during the study. Therefore, results are reported in terms of the nominal loading rates.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
The test medium (100 mg/L) was prepared by weighing the test substance (100 mg) onto a glass cover slip, which was then added to a volumetric flask (1 L) and made up to volume with dilution medium. The contents were stirred overnight in the dark before the stirrer was turned off and a siphon tube inserted at the mid-vessel point. The bulk medium was then left to stand overnight, after which the mid-vessel fraction was siphoned off and used in the test (nominal loading rate of 100 mg/L WAF).
Test organisms (species):
Pseudokirchneriella subcapitata (previous names: Raphidocelis subcapitata, Selenastrum capricornutum)
Details on test organisms:
TEST ORGANISM
- Common name: green algae
- Strain: CCAP 278/4
- Source (laboratory, culture collection): a reputable supplier
- Age of inoculum (at test initiation): three days
- Method of cultivation: The liquid slope cultures were stored in an illuminated refrigerator. Sterile algal nutrient medium was inoculated with cells aseptically removed from the slope culture; these primary liquid cultures (100 mL) were incubated for approximately three days in an orbital incubator under continuous illumination (nominal temperatures 21 to 25C) to provide an inoculum in the log phase of growth, characterised by a cell density of 1.8 x E6 cells/mL.
- Any deformed or abnormal cells observed: no
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
72 h
Post exposure observation period:
None
Hardness:
Not reported
Test temperature:
21.9 - 23.4 °C
pH:
7.78 - 9.84
Dissolved oxygen:
Not reported
Salinity:
Not applicable to a freshwater study.
Nominal and measured concentrations:
Nominal concentration = 100 mg/L
Measured concentrations were not determined due to the very low water solubility of the test substance.
Details on test conditions:
TEST SYSTEM
- Test vessel: conical flask
- Type: loosely plugged with a foam bung
- Material, size, headspace, fill volume: glass, each flask contained 100 mL of control or test culture
- Aeration: orbital shaker, oscillating at a nominal 130 - 150 cycles per minute.
- Renewal rate of test solution (frequency/flow rate): none, test conducted under static conditions
- Initial cells density: 1.16 x E4/mL.
- Control end cells density: 1636117 cells/mL
- Test end cells density: 1916478 cells/mL
- No. of vessels per concentration (replicates): 6
- No. of vessels per control (replicates): 6

GROWTH MEDIUM
- Standard medium used: yes

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: filtered, dechlorinated tap water which had been softened and treated by reverse osmosis, before microfiltration and purification (resistivity of 18 Megohm/cm).
- Resistivity: 18 Megohm/cm)

OTHER TEST CONDITIONS
- Sterile test conditions: yes
- Adjustment of pH: none
- Photoperiod: continuous
- Light intensity and quality: nominally 4440 to 8880 lux, provided by 6 x 30 W “cool white” 1 metre fluorescent tubes


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: electronic particle counter

TEST CONCENTRATIONS
The range finding test employed nominal test concentrations of 1.0, 10 and 100 mg/L. After 72 hours, no inhibition of growth occurred at 1.0, 10 or 100 mg/L. A definitive limit test was therefore carried out at a nominal loading rate of 100 mg/L.
Reference substance (positive control):
no
Duration:
72 h
Dose descriptor:
EL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
biomass
Duration:
72 h
Dose descriptor:
EL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Duration:
72 h
Dose descriptor:
EL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: yield
Duration:
72 h
Dose descriptor:
NOELR
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
biomass
Duration:
72 h
Dose descriptor:
NOELR
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Duration:
72 h
Dose descriptor:
NOELR
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: yield
Details on results:
No microscopic abnormalities of the cells were detected.

The mean coefficient of variation (CoV) for section by section daily growth rates in control cultures was 9.2 during the definitive test and the CoV for the average specific growth rates of control cultures was 2.13 during the 72 hour exposure period.

Due to low aqueous solubility of the test substance, it was not possible to develop a suitable method of analysis with sufficient sensitivity to verify concentrations during the study to determine the acute toxicity of Daphnia Magna (HLS Study number MZD0032). Therefore, test results have been reported in terms of nominal loading rates.
Validity criteria fulfilled:
yes
Remarks:
Specific growth rate of at least 0.92/day. Mean CoV for daily growth rates in controls <=35% and CoV average specific growth rate in controls <=7%.
Conclusions:
LZ1554 was not found to be acutely toxic to algae when dispersed in medium at a nominal loading rate of 100 mg/L.
Executive summary:

The acute algal toxicity was determined in a GLP-compliant study in accordance with OECD no. 201 and EC test method C.3. LZ1554 was not found to be acutely toxic to algae when dispersed in medium at a nominal loading rate of 100 mg/L.

Description of key information

Algae

LZ1554 was not found to be acutely toxic to algae (Pseudokirchneriella subcapitata) when dispersed in medium at a nominal loading rate of 100 mg/L. Hence, the 72 hour EL50 (biomass, growth rate, yield) as well as the NOELR were determined to be > 100 mg/L nominal.

Key value for chemical safety assessment

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

Additional information