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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:
2016-08-31 to 2016 12 21, with the definitive exposure phase from 2016-11-30 to 2016-12-21
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 211 (Daphnia magna Reproduction Test)
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
Determination of the test item
The limit concentration (pH-neutralized and un neutralized) were analyzed via LC-MS/MS once within 7 days in fresh media at the start of an exposure interval (0 hours) and in old media at the end of an exposure interval (48 or 72 hours). The method was validated prior to this study according to SANCO 3029/99 rev.4 (2000).

Sampling for the analytica monitoring
At the start of the exposure intervals, samples of the fresh media were taken after preparation of the limit concentration (pH-neutralized and un neutralized) and analyzed. At the end of the exposure intervals (48 or 72 hours), samples of the old media were taken directly from the test vessels. For the longest exposure interval of 72 hours samples were taken at the start (0 hours) and at the end of the exposure interval (72 hours) once within the test period.

Range (target)
Recoveries of the test item should be within ± 20% of the nominal or initially measured concentration.
Vehicle:
not specified
Details on test solutions:
Definitive Test
Based on the results of an acute immobilization test performed at the test facility (Noack Study-ID: 151014CS / DAI16839), the definitive reproduction test was carried out with a limit concentration of the test item Genamin DMG 75 of 133 mg/L (pH-neutralized and un-neutralized), prepared with test medium.

Preparation of the limit concentration
The limit concentration (133 mg/L of the test item was weighed out) was freshly prepared with dilution water for each exposure interval. The limit concentration was mixed thoroughly by manual agitation.

pH-neutralized concentration
The limit concentration 133 mg/L was initially adjusted to pH 7 ± 0.2 by the addition of 1 M HCl and tested under the same conditions as the un-neutralized concentration.

Control
10 replicates of dilution water without test item

Test method
The study was performed with a semi-static test design. A medium renewal 3 times per week (i.e. on Monday, Wednesday and Friday) was sufficient.

Renewal of thetest solutions
The test solutions were renewed 3 times per week (after 48 or 72 hours). For this purpose, a second set of test vessels was filled with the freshly prepared test solutions and the daphnids were transferred by pipette (see ‘Application’).

Test duration
21 days

Test vessels
Glass beakers (5 (ID) x 8 (H) cm), 100 mL capacity, loosely covered with watch glasses

Test volume
50 mL

Dilution water
Same as the culture medium, therefore no acclimation necessary

Number of daphnids and replicates
10 daphnids in 10 replicates were used for the limit concentration (pH-neutralized and un-neutralized) and the control with one daphnid per replicate, which was held individually.

Age of the daphnids at the start of the exposure
Less than 24 hours old daphnids from a healthy stock were used for the study. Juvenile daphnids were removed from the culture vessels at the latest 24 hours before the start of the exposure and discarded. The juveniles born within this period of max. 24 hours preceding the exposure were used for the test. No first brood progeny was used for the test.

Application
50 g test solution per replicate was weighed out into the test vessels. This corresponds to 50 mL. The daphnids were transferred in a small amount of dilution water or test solution by pipette.

Feeding
Daily feeding per test vessel; Pseudokirchneriella subcapitata (0.889 - 1.35 mL) and Desmodesmus subspicatus (0.650 -0.877 mL) suspension was provided as food corresponding to 0.2 mg C per Daphnia and day.
There was variation according to the density of the algae suspension, but it was the same for all test groups on each feeding day.

Test temperature (target)
20 ± 1 °C

Light intensity (target)
Max. 1500 lx

Photoperiod
16/8 hours light/dark cycle

Aeration
Test vessels were not aerated during the test.
Test organisms (species):
Daphnia magna
Details on test organisms:
Test system Daphnia magna STRAUS (Clone 5)

Reason for the selectionof the test system
Daphnia magna is the preferred species in accordance with the
test guideline and is bred at the test facility.
The Daphnia magna culture of Clone 5 has shown to meet the validity criteria for the reproduction test of a mean of ≥ 60 living juveniles per survived parent animal when cultured under the described conditions.

Origin
Institut für Wasser-, Boden- und Lufthygiene (WaBoLu),
14195 Berlin, Germany

Breeder
Noack Laboratorien GmbH,
Käthe-Paulus-Str. 1, 31157 Sarstedt, Germany

Culture
In glass vessels (2 - 3 L capacity) with approximately 1.8 L culture medium, at 20  2 °C, in an incubator, 16 hours illumination; light intensity of max. 20 µEm-2  s-1 (max. 1340 lx)

Culture medium
Elendt M4, according to OECD 202, Annex 3 (2004), modified to a total hardness of 160 to 180 mg CaCO3/L, is used. The composition of the culture medium is presented

Culture feeding
The culture daphnids are fed at least 5 times per week ad libitum with a mix of unicellular green algae, e.g. Pseudokirchneriella subcapitata and Desmodesmus subspicatus, with an algae cell density of > 106 cells/mL. The algae are cultured at the test facility.

Origin of the food algae
Sammlung von Algenkulturen (SAG), Pflanzenphysiologisches Institut der Universität Göttingen, Nikolausberger Weg 18, 37073 Göttingen, Germany
Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
21 d
Hardness:
Total Hardness [mg/L] as CaCO3
Nominal
test item concentrations
[mg/L] fresh old fresh old fresh old
Day 0 Day 2 Day 14 Day 16 Day 16 Day 19
2016-11-30 2016-12-02 2016-12-14 2016-12-16 2016-12-16 2016-12-19
133 (pH-neutralized) 174 163 168 163 174 169
133 (un-neutralized) 180 164 169 160 176 164
Control 174 172 171 164 176 173
The total hardness was above 140 mg/L as CaCO3.
Test temperature:
Temperature [°C]
Nominal
test item concentrations
[mg/L] fresh old fresh old fresh old
Day 0 Day 2 Day 14 Day 16 Day 16 Day 19
2016-11-30 2016-12-02 2016-12-14 2016-12-16 2016-12-16 2016-12-19
133 (pH-neutralized) 20.2 20.3 20.2 20.2 20.3 20.0
133 (un-neutralized) 20.1 20.4 20.2 20.2 20.4 20.1
Control 19.9 20.4 20.9 20.2 20.9 20.0
The temperature was within the range of 20 ± 1 °C.
pH:
pH-Values
Nominal
test item concentrations
[mg/L] fresh old fresh old fresh old
Day 0 Day 2 Day 14 Day 16 Day 16 Day 19
2016-11-30 2016-12-02 2016-12-14 2016-12-16 2016-12-16 2016-12-19
133 (pH-neutralized) 6.92 7.50 7.17 7.09 6.99 7.30
133 (un-neutralized) 9.40 7.65 9.39 7.98 9.26 7.94
Control 7.56 7.19 8.63 7.84 8.10 7.56
The pH-values should be in the range of 6 – 9. Significant deviations ≥ 1.5 units between the initial and final pH-values were not observed in the control and in the pH-neutralized limit concentration. However, the initial pH-value of the un-neutralized limit concentration was > pH 9 and the deviation between the initial and final pH-value was > 1.5 units in the first exposure interval. Since initial pH-values of > 9 were expected, the limit concentration was additionally tested pH-neutralized.
Dissolved oxygen:
Dissolved Oxygen Concentration [mg/L]
Nominal
test item concentrations
[mg/L] fresh old fresh old fresh old
Day 0 Day 2 Day 14 Day 16 Day 16 Day 19
2016-11-30 2016-12-02 2016-12-14 2016-12-16 2016-12-16 2016-12-19
133 (pH-neutralized) 9.18 9.55 8.99 7.04 9.36 8.75
133 (un-neutralized) 9.32 8.38 8.80 7.21 9.29 8.68
Control 9.57 7.91 9.28 7.27 9.54 8.64
The dissolved oxygen concentration was above 3 mg/L.
Details on test conditions:
see detail on test solutions
Reference substance (positive control):
yes
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
>= 133 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Duration:
21 d
Dose descriptor:
LOEC
Effect conc.:
> 133 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Results with reference substance (positive control):
The percentage of immobility for the reference item potassium dichromate (SIGMA-ALDRICH, batch number MKBV0900V, purity 99.0%, CAS RN 7778-50-9) was determined after 24 hours from 2016 11-24 to 2016-11-25.

EC50-Value (with 95% confidents limits) of the Reference Item Potassium dichromate
based on nominal concentrations mg/L, (0 - 24 hours)
Current Study Valid Range
EC50 0.802 mg/L
0.6 - 2.4 mg/L, acc. to AQS P 9/2 (02/2000); clone 5
0.6 - 2.1 mg/L, acc. to OECD 202 (2004); clone A
95% confidence limits 0.597 - 0.942 mg/L


EC-values and statistical analyses for reference item
The EC50-value for the reference item was calculated by sigmoidal dose-response regression. The respective 95% confidence limits were calculated from the standard error and the t-distribution. All calculations were carried out from the best-fit values with the software GraphPad Prism5.
Reported statistics and error estimates:
Statistical evaluation
The NOEC and LOEC for the reproduction were determined with the software ToxRat Professional as specified below.
Significant deviations of reproduction were determined in comparison to the control using statistical standard procedures as normality test (Shapiro-Wilk’s test), variance homogeneity test (Levene’s test), trend analysis (Trend analysis by contrasts (monotonicity of concentration-response) and a multiple test (William’s test; a one-sided test for reduction of the reproductive output and a two-sided for the stimulation of reproduction ).
P values of the normality and the variance homogeneity test were 0.01 and 0.05 for the trend analysis. The -value (acceptable probability of incorrectly concluding that there is a difference) was  = 0.01 for the normality test and the variance homogeneity test and  = 0.05 for the multiple test.
For the effects on reproduction, the juveniles per survived parent excluding accidental and inadvertent mortality (reproductive output) were calculated. Since no accidental and inadvertent mortality was observed, no parental daphnids were excluded from the evaluation of the reproductive output. Since all parental daphnids survived until the end of the study, the number of survived parent animal is the same as for the introduced parental daphnids and therefore no additional evaluation per introduced parental daphnids is necessary.
Since no mortality appeared in this study, statistical evaluation for the adult mortality was not necessary.
The coefficients of variation around the mean number of living offspring produced per introduced parent and per survived parent in the control and the test groups were evaluated.
The NOEC and the LOEC given are based on the nominal test item concentration, since the measured test item concentrations were all within ± 20% of the nominal concentration.





The number of juveniles in all replicates of the control and the test groups was counted and recorded every day.

The average number of living juveniles at the end of the test after 21 days per survived parental daphnid was 113 in the control group and thus thevalidity criterion for the reproduction of a mean of ≥ 60 living offspring per survived parent animalin the control was met.

No parental daphnids were excluded due to accidental or inadvertent mortality from the evaluation of the reproductive output. No statistically significant reduction of the reproductive output in comparison to the reproductive output in the control was determined at the limit concentration 133 mg/L (pH-neutralized and un-neutralized). Both test item concentrations (pH-neutralized andun-neutralized) caused a significant stimulating effect (approx. 60% stimulation) on the reproduction of the daphnids. This stimulating effect is considered to be not an adverse effect.

 

Effects on Reproduction for all parental Daphnids (based on the nominalconcentration ofGenamin DMG 75)

Nominal

test item concentration

Mean number of offspring per survived / introduced parental daphnid

[mg/L]

Mean

SD

CV

133 (pH-neutralized)

181

17.1

 9.5

133 (un-neutralized)

184

25.2

13.7

Control

113

17.0

15.0

 

 

 Adult Mortality

 

Adult mortality was not observed in this study. Therefore, no statistical evaluation was carried out for this parameter.

 

Mortality [%] of the Adult Daphnids after 7, 14 and 21 Days of Exposure

                (n = 10)

Nominal

test item concentrations

[mg/L]

Adult Mortality [%]

7 days

14 days

21 days

133 (pH-neutralized)

0

0

0

133 (un-neutralized)

0

0

0

Control

0

0

0

 

 

  Intrinsic Rates of Natural Increase (IR)

 

Theintrinsic rates of natural increase (IR)of the parental daphnids accounting for generation time and number of offspring were used for calculation of population growth and maintenance. For details, seeTable5.

 

  Intrinsic Rates of Natural Increase

Nominal

test item concentrations

[mg/L]

Intrinsic rate of natural increase in replicate no.

Mean IR

CV

1

2

3

4

5

6

7

8

9

10

MV±SD

[%]

133 (pH-neutralized)

0.421

0.503

0.403

0.405

0.440

0.437

0.421

0.400

0.413

0.460

0.431±

0.0318

 7.4

133 (un-neutralized)

0.473

0.459

0.467

0.422

0.475

0.418

0.408

0.453

0.410

0.457

0.444±

0.0267

 6.0

Control

0.454

0.365

0.441

0.363

0.341

0.471

0.427

0.344

0.388

0.374

0.397±

0.0474

11.9

Number of Broods and first Appearance of living Juveniles

 

At the end of the test after 21 days, the number of broods with living juveniles produced by the parental daphnids was 4 in the control. However, one parental daphnid in the control produced 5 broods with living juveniles. In the pH-neutralized limit concentration of 133 mg/L 5 broods with living juveniles were produced by the parental daphnids. In the un-neutralized limit concentration of 133 mg/L, two parental daphnids produced 4 broods with living juveniles and eight parental daphnids produced 5 broods.
The
first appearance ofliving juveniles was observed on days 7 to 10 at all parental daphnids of the control and the limit concentration 133 mg/L(pH-neutralized and un‑neutralized).

 

 First Appearance of Living Juveniles and Mean Number of Broods in the Individual Groups

Nominal

test item concentrations

[mg/L]

Day of first appearance of living juveniles at the parental daphnid in replicate no.

First

Mean

appearance

number

1

2

3

4

5

6

7

8

9

10

mean day

of broods

133 (pH-neutralized)

8

7

9

9

 9

9

9

 9

9

8

8.6

5.0

133 (un-neutralized)

8

8

8

9

 8

9

9

 8

9

9

8.5

4.8

Control

8

9

9

9

10

7

9

10

9

9

8.9

4.1

 

 

 Stillborn Juveniles and Aborted Eggs

 

Nostillborn juveniles or aborted eggs were observed in the control and in the limit concentration 133 mg/L(pH-neutralized and un‑neutralized) during the exposure period of 21 days.

 

 Stillborn Juvenilesand Aborted Eggs produced by the Parental Daphnids after 21 Days

Nominal

test item concentrations

[mg/L]

Number of

Total no.

 

å

Number of
adults

N

stillborn

juveniles

aborted

eggs

133 (pH-neutralized)

0

0

0

10

133 (un-neutralized)

0

0

0

10

Control

0

0

0

10

N   = number of the parental daphnids

 

  Relative Number of Dead (Stillborn + Aborted Eggs)to Total Number of Juvenilesper Parental Daphnids

Nominal

test item concentrations

[mg/L]

Number of juveniles

Percentage of

dead juveniles#)

[%]

Dead

Alive

Total

133 (pH-neutralized)

0

1807

1807

0

133 (un-neutralized)

0

1843

1843

0

Control

0

1133

1133

0

Dead   = aborted eggs + stillborn juveniles

Total  = dead + alive juveniles

#)          = related to the total number of juveniles

 Growth (Total Body Length and Mean Dry Weight) of the Parental Daphnids

 

The parental daphnids of the test concentrations were slightly larger and heavier than the animals of the control group.

The mean values of the body length (excluding the anal spine) of the parental daphnids in the pH‑neutralized and un‑neutralizedlimit concentration 133 mg/L were 4.93 mm per daphnid and 4.88 mm per daphnid in the control group.

The mean values of the dry weight of the parental daphnids were 0.86 and 0.97 mg per daphnid in thelimit concentration 133 mg/L (pH-neutralized and un‑neutralized)and 0.67 mg per daphnid in the control .

 

  Total Body Length and Mean Dry Weight of the Parental Daphnids

Nominal

test item concentrations

[mg/L]

Total length of the parent animals [mm]

MV

N

Mean
dry weight

[mg]

Replicate no.

 

 

1

2

3

4

5

6

7

8

9

10

[mm]

 

S

MV

133 (pH-neutralized)

5.00

5.00

5.00

4.75

5.00

5.00

4.75

5.00

4.75

5.00

4.93

10

9.7

0.97

133 (un-neutralized)

5.00

5.00

5.00

4.75

5.00

4.75

5.00

5.00

4.75

5.00

4.93

10

8.6

0.86

Control

5.00

4.75

5.00

5.00

4.75

4.75

5.00

5.00

4.75

4.75

4.88

10

6.7

0.67

N  = number of the parental daphnids

  

 Presence of Males

 No males were observed in the control or in the test groups during the test.

  

 Occurrence of Ephippia (Winter Eggs)

 No ephippiawere observedin the control or in the test groups during the test.

Observations in the Control

Date

(2016)

01.12.

02.12.

03.12.

04.12.

05.12.

06.12.

07.12.

08.12.

09.12.

10.12.

11.12.

12.12.

13.12.

14.12.

15.12.

16.12.

17.12.

18.12.

19.12

20.12.

21.12.

 

Day

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

Replicate No.

Number of Juveniles

total

1

23

36

15

12

19

 105

2

14

25

37

10

   86

3

25

44

48

30

 147

4

13

26

33

29

 101

5

18

 2

29

30

28

 107

6

14

30

33

7

28

 112

7

23

41

 6

28

16

 5

 119

8

19

 1

 1

39

27

24

 111

9

25

13

32

26

38

 134

10

15

31

34

31

 111

Based on all survived / introduced parental daphnids:

1133

Mortality of Parental      (N)

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

     0

Mortality of Parental     [%]

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

     0

 

     = no juveniles observed

 


 

Observations in the Nominal Test Item Concentration of 133 mg/L (pH-neutralized)

Date

(2016)

01.12.

02.12.

03.12.

04.12.

05.12.

06.12.

07.12.

08.12.

09.12.

10.12.

11.12.

12.12.

13.12.

14.12.

15.12.

16.12.

17.12.

18.12.

19.12

20.12.

21.12.

 

Day

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

Replicate No.

Number of Juveniles

total

1

12

38

40

41

39

 170

2

16

48

50

35

27

 176

3

22

35

43

32

26

 158

4

21

 1

40

 1

45

 5

26

28

 167

5

26

48

41

40

19

 2

 176

6

22

52

45

58

37

 214

7

28

39

 1

47

41

34

 190

8

18

38

57

46

35

 194

9

25

42

40

28

32

 167

10

24

40

50

45

36

 195

Based on all survived / introduced parental daphnids:

1807

Mortality of Parental      (N)

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

     0

Mortality of Parental     [%]

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

     0

 

     = no juveniles observed

 

 


 

Observations in the Nominal Test Item Concentration of 133 mg/L (un-neutralized)

Date

(2016)

01.12.

02.12.

03.12.

04.12.

05.12.

06.12.

07.12.

08.12.

09.12.

10.12.

11.12.

12.12.

13.12.

14.12.

15.12.

16.12.

17.12.

18.12.

19.12

20.12.

21.12.

 

Day

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

Replicate No.

Number of Juveniles

total

1

24

58

54

50

 186

2

22

50

54

41

27

 194

3

23

57

49

41

34

 204

4

28

44

49

33

25

 179

5

25

 2

53

51

38

32

 201

6

28

35

1

52

29

 145

7

21

44

47

31

 143

8

 5

25

46

54

42

22

18

 212

9

22

42

1

51

32

21

 169

10

30

58

51

39

30

 2

 210

Based on all survived / introduced parental daphnids:

1843

Mortality of Parental      (N)

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

     0

Mortality of Parental     [%]

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

     0

 

     = no juveniles observed

Validity criteria fulfilled:
yes
Conclusions:
Since all parental daphnids survived until the end of the study, the effects on reproduction and other sub-lethal effects were evaluated based on the reproduction per survived parent animal, which is equivalent to the introduced parental daphnids.
The overall effect threshold for effects of the test item under the test conditions was ≥ 133 mg/L (NOEC) and > 133 mg/L (LOEC) based on the nominal concentration of the test item Genamin DMG 75. These values are based on inhibition. Both test item concentrations (pH-neutralized and un-neutralized) caused a significant stimulating effect (approx. 60% stimulation) on the reproduction of the daphnids.
Executive summary:

A Daphnia magna reproduction test (semi-static, 21 d) of the test item Genamin DMG 75 (batch number:RAK-KRS-00022) was conducted at the test facility according to OECD 211 (2012) from 2016-08-31 to 2016‑12‑21, with the definitive exposure phase from 2016-11-30 to 2016-12-21.

Test species was Daphnia magna STRAUS (Clone 5). Ten daphnids, held individually, were used per concentration level and control. At test start the daphnids were less than 24 hours old. The study was carried out under semi-static conditions with a medium renewal three times per week. Aim of the test was to assess the effects on the reproduction capacity and other sub-lethal effects.

The test item Genamin DMG 75 is a slightly yellowish solution, which is soluble in water. The study was performedas a limit test with the limit concentration of the test item Genamin DMG 75 of 133 mg/L (pH-neutralized and un-neutralized). Since the test item induced high pH-values in aqueous solution (> pH 9), the limit concentration of 133 mg/L was tested pH-neutralized in addition to the un-neutralized concentration. The pH-neutralization by addition of 1 M HCl reflects the actually existing environmental conditions in the buffered aquatic systems.

The test item concentrations of Genamin DMG 75 were analytically verified via LC-MS/MS in fresh media at the start of the exposure intervals on days 0, 12, 16 (0 hours) and in old media at the end of the exposure intervals on days 2, 14, 19 (48 or 72 hours, respectively) in the limit concentration (pH-neutralized and un-neutralized) and the control.

The measured concentrations of the test item in fresh media at the start of the respective exposure intervals (0 hours) were in the range of 103 to 110% of the nominal concentration. At the end of the respective exposure intervals (48 and 72 hours), the measured concentrations in old media were in the range of 96 to 101% of the nominal concentration. The measured test item concentrations were all within±20% of the nominal concentration. Therefore,the nominal concentration of the test item Genamin DMG 75 was used for the evaluation of the effect concentrations.

The environmental conditionswere within the acceptable limits.The validity criteria of the test guideline were met.

Since all parental daphnids survived until the end of the study, the effects on reproduction and other sub-lethal effects were evaluated based on the reproduction per survived parent animal, which is equivalent to the introduced parental daphnids. A summary of all endpoints based on the nominal concentration of the test item GenaminDMG 75 is given below.

 

 Endpoints for Reproduction (based on the nominal test item concentration)

Effect concentration

Genamin DMG 75

Nominal test item concentration
[mg/L]

LOECReproduction

> 133

NOECReproduction

133

 

The values in above are based on inhibition. Both test item concentrations (pH-neutralized andun-neutralized) caused a significant

stimulating effect (approx. 60% stimulation) on the reproduction of the daphnids.The parental daphnids of the test concentrations were slightly larger and heavier than the amimals of the control group.

Description of key information

Since all parental daphnids survived until the end of the study, the effects on reproduction and other sub-lethal effects were evaluated based on the reproduction per survived parent animal, which is equivalent to the introduced parental daphnids.

The overall effect threshold for effects of the test item under the test conditions was ≥ 133 mg/L (NOEC) and > 133 mg/L (LOEC) based on the nominal concentration of the test item Genamin DMG 75. These values are based on inhibition. Both test item concentrations (pH-neutralized and un-neutralized) caused a significant stimulating effect (approx. 60% stimulation) on the reproduction of the daphnids.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates
Effect concentration:
133 mg/L

Additional information