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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:
March 2013-September 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study, GLP
Qualifier:
according to guideline
Guideline:
OECD Guideline 211 (Daphnia magna Reproduction Test)
Version / remarks:
"Daphnia magna Reproduction Test", dated May 30, 2008, adopted October 03, 2008 and adopted October 02, 2012.
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
One sample from the freshly prepared stock solution and duplicate samples from the freshly prepared test media of all test concentrations and the
control were taken at the first treatment period (Day 0) and at a treatment period in the second and third week (Days 7 and 16). For the determination of the stability of the test item under the test conditions and of the maintenance of the test item concentrations during the renewal periods, duplicate samples of the respective aged test media and controls were sampled at Day 2, 9 and 19. One of these three stability control treatments lasted for 72 hours (weekend), two for 48 hours, corresponding to the different test medium renewal periods. All samples were diluted by a factor of two with
acetonitrile directly after sampling. All samples were centrifuged (4000 rpm, 10 minutes).
Storage of Samples: All samples were measured immediately after sampling. Spare samples were stored in the freezer (at ≤ - 10 °C) until finalisation of the study.
Fortified Samples: Approximately 50 mg test item were diluted with 50 mL acetonitrile to obtain two stock solutions of 1 g test item/L each. Appropriate amounts of these stock solutions were diluted with test water to obtain fortified samples at concentrations of 0.08, 0.1, 0.25 and 2.5 mg test item/L. Exact values were documented in the raw data.
Vehicle:
no
Details on test solutions:
Before test start and before every test medium renewal, a concentrated stock solution of nominal 40 mg/L was prepared by mixing 42.8 µL test item into 1000 mL test water by intense stirring for 10 minutes. Adequate volumes of this test medium were diluted with test water to prepare the test media. The test media were prepared just before introduction of Daphnia (= start of the test) and each test medium renewal.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: water flea
- Strain/clone: Daphnia magna (Straus), clone 5
- Sex: female
- Source: in-house laboratory culture
- Age of parental stock: 3 to 15 hours old
- Feeding during test
- Food type: green algae (Desmodesmus subspicatus), freshly grown in the laboratory
- Frequency: at least all working days

ACCLIMATION
not necessary since the breeding is performed in the same kind of test water
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
21 d
Hardness:
2.5 mmol/L (= 250 mg/L) as CaCO3
Test temperature:
19 - 20 °C in the freshly prepared media; 20 °C in the aged media
pH:
7.5 to 8.1 in the freshly prepared media; 7.2 to 8.9 in the aged media; and thus the pH-value did vary by more than 1.5 units
Dissolved oxygen:
8.2 to 8.9 mg/L in the freshly prepared media; 7.9 to 10.0 mg/L in the aged media
Nominal and measured concentrations:
nominal: 2.0, 1.0, 0.50, 0.25 and 0.125 mg test item/L and a control.
Details on test conditions:
Details on test conditions:
TEST SYSTEM
- Test vessel: Glass beakers of approximately 100 mL volume which were filled completely (i.e. remaining head space reduced to a technical possible minimum) with test medium and kept closed with a conical glass stopper
- Renewal rate of test solution (frequency/flow rate): Semi-static, The test media of all test concentrations and of the control were renewed on Days 2, 5, 7, 9, 12, 14, 16 and 19 of the exposure period (every Monday, Wednesday and Friday). At these dates the test animals were carefully transferred from the old test vessels into the freshly prepared test media of the corresponding concentrations. The handling and the allocation of the treatments was randomised on every test media renewal- 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: Elendt M4, modified to a total hardness of 2.5 mmol/L (= 250 mg/L) as CaCO3 .
- Intervals of water quality measurement: The pH-values, dissolved oxygen concentrations and water temperatures were measured in all test concentrations and the control in all freshly prepared and aged test media.
OTHER TEST CONDITIONS
- Photoperiod: 16/8 h light/dark cycle
- Light intensity: 1000 - 1300 lux.
- Test temperature: 19 - 20 °C in the freshly prepared media; 20 °C in the aged media;

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Mortality of parental daphnids (immobile or dying) – daily
Condition of parental daphnids:
Abnormalities (e.g. swimming behaviour, number of males and winter eggs) were observed and recorded at each day of observation.
Total length and dry body weight of parental daphnids:
At the end of the test total length of each daphnid and dry body weight of the daphnids alive at each concentration level and the control was determined.
Number of offspring (alive and dead progeny) - daily
neonates (alive and dead) were removed after counting
dead specimens were removed

Reference substance (positive control):
no
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
0.125 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Remarks on result:
other: acc. to 2012 version of OECD 211
Duration:
21 d
Dose descriptor:
LOEC
Effect conc.:
0.25 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Remarks on result:
other: acc. to 2012 version of OECD 211
Duration:
21 d
Dose descriptor:
EC50
Effect conc.:
1.46 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Remarks on result:
other: 1.15-2.03 mg/l
Key result
Duration:
21 d
Dose descriptor:
EC10
Effect conc.:
0.272 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Remarks on result:
other: 0.130-0.402 mg/l
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
0.25 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
21 d
Dose descriptor:
LOEC
Effect conc.:
0.5 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Reported statistics and error estimates:
see section "Any other information on results"

Effects

Nominal concentration of Allyl methacrylate
 [mg/L]

 

Control

0.125

0.25

0.5

1.0

2.0

Mean Number of Living Juveniles per

Producing Parent (Reproduction Rate±SD)

175.2±16.2

165.2±14.9

155.0±8.3

156.9±13.3

143.4±9.1)

-

Coefficient of Variation of theMean Number of Juveniles per Producing Parent [%]

9.2

9.0

5.3

8.5

6.3

-

Percentage of Live Juveniles Related to the Total Number of Juveniles [%]

-

94.3

88.5

89.5

81.9

-

Adult surviving after 21 Days [%]

-

100

100

70* 0

70* 0

0

- = adult died in the course of the study

**: significantly different from the control (tested with Williams t-test, α=0.05, one sided)

Validity criteria fulfilled:
yes
Conclusions:
The reduction of the reproductive output was the most sensitive effect in this study. The No Observed Effect Concentration (NOECReproduction) after 21 days was assessed at 0.1259 mg/L.. The EC10-value for the reduction of the reproductive output was 0.272 mg/L (95 % confidence limits: 0.130 -0.402 mg/L). The EC50-value for the reduction of the reproductive output was 1.46 mg/L (95 % confidence limits: 1.15 2.03 mg/L).
The NOEC for the adult mortality was 0.25 mg/L. The LOEC was 0.5 mg/L. All values based on nominal concentrations.
Executive summary:

The 21-day-chronic toxicity of Allyl methacrylate to Daphnia magna Straus was studied under semi static conditions according to OECD guideline 211 (adopted 3 October 2008 and adopted 2 October 2012). Daphnids were exposed to control and test chemical at nominal concentrations of 0 (control),0.125, 0.25, 0.5, 1.0 and 2.0 mg/L

The 21 d EC10 based on reproduction was 0.272mg/L(95 % c.l. 0.130 – 0.402 mg/L).

The 21d NOEC based on reproduction was 0.125 mg/L. (nominal concentrations).The 21 d EC50 based on reproductive effects was 1.46 mg/L(95 % c.l. 1.15 – 2.03 mg/L).The 21 d NOEC based on mortality was 0.25 mg/L.

The sublethal effects included were reproduction rate and appearance of first brood. Production of offspring in the treated groups indicated that Allyl methacrylate had an effect on the reproduction at concentrations greater than 0.125 mg/L. The most sensitive end point was the reduction of the reproductive output.

Results Synopsis 

Test Organism Age (e.g. 1st instar): 3 to 15 h old daphnids

Test Type: Semi static

 

NOEC: 0.125 mg/L

EC10: 0.272 mg/L95 % confidence limits: 0.130 – 0.402 mg/L

LOEC: 0.25 mg/L

Endpoint(s) Effected: reproduction

 

 

Description of key information

The 21 d EC10 of Allyl methacrylate in freshwater invertebrates based on reproduction was 0.272 mg/L. The 21 d-NOEC based on reproduction was 0.125 mg/L (OECD guideline 211, semi static conditions).

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates
Effect concentration:
0.272 mg/L

Additional information

One long-term study in freshwater invertebrates is available for Allyl methacrylate which is considered reliable without restriction (Klimisch score 1).

The 21-day-chronic toxicity of Allyl methacrylate to Daphnia magna Straus was studied under semi static conditions according to OECD guideline 211 (adopted 3 October 2008 and adopted 02 October 2012). Daphnids were exposed to control and test chemical at nominal concentrations of 0 (control), 0.125, 0.25, 0.5, 1.0 and 2.0 mg/L.

The 21 day NOEC based on mortality was 0.25 mg/L. The 21 day EC50 based on reproductive effects was 1.46 mg/L(95 % c.l. 1.15 – 2.03 mg/L). The 21 -d EC10 based on reproduction was 0.272mg/L(95 % c.l. 0.130 – 0.402 mg/L).The 21-day NOEC based on reproduction was 0.125 mg/L. (nominal concentrations).

The sublethal effects included were reproduction rate and appearance of first brood. Production of offspring in the treated groups indicate that Allyl methacrylate had an effect on the reproduction at concentrations greater than 0.125 mg/L. The most sensitive end point was the reduction of the reproductive output.