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Long-term toxicity to aquatic invertebrates

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Reference
Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Feb 13 - Mar 06, 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study with analytical monitoring
Qualifier:
according to
Guideline:
OECD Guideline 211 (Daphnia magna Reproduction Test)
Version / remarks:
3 October, 2008
Deviations:
yes
Remarks:
The pH values (7.77– 9.91) outside recommended range of 6 – 9, deviation > 1.5 units at all treatment levels. The temperature was 18.1 – 20.9°C, variance < 2°C was not maintained. This had no effect on reproduction or the test concentrations.
GLP compliance:
yes (incl. certificate)
Remarks:
Ministerium für Arbeit, Integration und Soziales des Landes Nordrhein-Westfalen, Düsseldorf, 07 Feb 2011
Analytical monitoring:
yes
Details on sampling:
- Concentrations: all concentrations
- Sampling method: sampled from representative fresh (0 h) and aged (24 h) test solutions three times per week from pooled solutions
- Sample storage conditions before analysis: immediately analysed or stored in a freezer at -25°C until analysis
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The test substance was directly dissolved with 2 hours stirring. An aliquot of the test item (purity 73.3%) was weighed onto a sterile Teflon plate which was then placed directly into a sterile 2 L Schott bottle. Under laminar flow 2 L of sterile dilution water was added to the mixing vessels to reach a loading corresponding to the highest test concentration of 12 mg a.s./L.
The content of the vessel was stirred at 300 rpm for 2 h at room temperature (21 ± 1°C). The eluate was then used to begin a 50% serial dilution for the preparation of the remaining four test concentrations using sterilised dilution water.
The eluate was prepared 2 h under sterile conditions and replaced daily.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: water flea
- Source: German Federal Environment Agency, Institut für Wasser-, Boden- und Lufthygiene. Specimens used in the test were bred in the laboratory of the Fraunhofer IME
- Age of parental daphnids: at least 3 weeks old
- Feeding during test: yes
- Food type: with concentrated suspensions of unicellular alga Desmodesmus subspicatus
- Amount: of 0.2 mg carbon/L per daphnid per day
- Frequency: at each media renewal (daily)

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES: . Newborn Daphnia (between 4 and 23 hours old) were removed by wide-bore pipette (to avoid damage) and isolated in fresh dilution water for at least 1 hour prior to being added randomly to the test vessels containing the appropriate test or control media.
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
21 d
Hardness:
1.0 mmol/L
Test temperature:
18.1 - 20.9 °C
pH:
7.77 - 9.91
Dissolved oxygen:
6.8 - 11.0 mg/L
Nominal and measured concentrations:
Nominal test concentrations: control, 0.51, 1.13, 2.48, 5.45, 12 mg/L
Mean measured test concentrations (TWA): -, 0.45, 1.03, 2.33, 5.27, 11.75 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: glass beakers
- Material, nominal volume, fill volume: glass, 100 ml, 60 ml
- Aeration: no
- Renewal rate of test solution: daily
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates): 10

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Cu-free tap water according to OECD guideline 211 sourced from the Schmallenberg district water production plants, mostly fed by small springs and percolation. The purification process occurs on site at Fraunhofer IME and includes filtration with activated charcoal, passage through a lime-stone column, and aeration to the point of oxygen saturation.
- Alkalinity: 1.3-1.5 mmol/L
- Conductivity: 203.7 - 227.9 µS/cm
- Culture medium different from test medium:- Intervals of water quality measurement:

OTHER TEST CONDITIONS
- Photoperiod: light/dark cycle of 16/8 hours
- Light intensity: 800 lx

EFFECT PARAMETERS MEASURED: The numbers of immobile daphnids were visually determined daily and the immobile daphnids were removed. Any abnormalities in appearance and behaviour were recorded. The newborn daphnids per beaker were counted and removed daily until day 10 (after day 10 newborn daphnids were counted and removed daily at each water renewal), abnormalities in condition (including male sex) or presence of winter eggs were checked and recorded. At study termination, length of the adults up to the highest treatment without significant mortality were measured by digital photography and image analysis and compared with the equally measured control animals.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.2
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
5.3 mg/L
Nominal / measured:
meas. (TWA)
Conc. based on:
act. ingr.
Basis for effect:
reproduction
Duration:
21 d
Dose descriptor:
LOEC
Effect conc.:
11.75 mg/L
Nominal / measured:
meas. (TWA)
Conc. based on:
act. ingr.
Basis for effect:
reproduction
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
>= 11.75 mg/L
Nominal / measured:
meas. (TWA)
Conc. based on:
act. ingr.
Basis for effect:
mortality
Remarks:
parental daphnids
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
>= 11.75 mg/L
Nominal / measured:
meas. (TWA)
Conc. based on:
act. ingr.
Basis for effect:
growth
Remarks:
(body length)
Details on results:
- Mortality of parent animals: One parental daphnid died in the third highest treatment (2.33 mg/L; TWA) on day one and two animals died on day three in the second highest treatment (5.27 mg/L; TWA)
- Type and number of morphological abnormalities: none
- Type and number of behavioural abnormalities: none

Table 1: Validity criteria for OECD 211.

Criterion from the guideline

Outcome

Validity criterion fulfilled

The mortality of the parent animals in the control (female Daphnia) does not exceed 20% at the end of the test.

 0%

 yes

The mean number of living offspring produced per parent animalsurvivingat the end of the test is ≥ 60 in the control.

 65.4

 yes

Validity criteria fulfilled:
yes
Remarks:
Please see Table 1 at "Any other information on results incl. tables"

Description of key information

NOEC (21 d) = 5.3 mg a.i./L  (measured, TWA) related to reproduction.

Key value for chemical safety assessment

EC10, LC10 or NOEC for freshwater invertebrates:
5.3 mg/L

Additional information

The chronic toxicity of the substance to aquatic invertebrates was tested in a GLP guideline study conducted according to OECD 211. Cu-free tap water according to OECD guideline 211 sourced from the Schmallenberg district water production plants was used as test medium. Juvenile Daphnia magna were exposed the test substance in a semi-static test system for 21 days. The nominal test substance concentrations were 0.51, 1.13, 2.48, 5.45, and 12 mg/L. The substance concentrations in freshly prepared and aged test solution were analytically verified by liquid chromatography (LC) and tandem mass spectrometry detection (LC-MS/MS). The time weighted average (TWA) concentrations were 0.45, 1.03, 2.33, 5.27, and 11.75 mg/L. The effects on growth and survival as well as reproductive performance were investigated in the test. The substance did not affect survival, growth or time of first brood of adult Daphnia. A concentrations related effect on the reproduction rate was observed. The determined NOEC was 5.3 mg/L (measured, TWA) based on reproduction.