REAKTIV-GELB F-68 072 FW

Long-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

05 May 2006 to 19 July 2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method C.20 (Daphnia magna Reproduction Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Stability of test concentration/s during exposure: Ensured by renewal of test media three times per week and verified by chemical analysis (HPLC) according to the sampling scheme described below.

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 0.1, 0.316, 1.0, 3.16 and 10 mg/l
- Sampling method: Daily by direct extraction, from each test media.
- Sample storage conditions before analysis: Routinely, the samples are analysed immediately. Only in exceptional cases, they are stored overnight deeply frozen and protected from light.

Vehicle:

no

Details on test solutions:

A stock solution was prepared to give the desired series of test concentrations. To achieve this approx. 40 mg of the test item were added to 2 litres of dilution water, and diluted accordingly.

Test organisms (species):

Daphnia magna

Details on test organisms:

Name: Daphnia magna STRAUS, parthenogenetic females
Source: Strain of Bundesgesundheitsamt Berlin
Maintenance and Acclimatisation: A population of parthenogenetic females of synchronized
age structure has been maintained for more than 15 years in the test facility under constant temperature conditions (20 +/- 1 °C) at a 16 : 8 hour light-darkness photoperiod (illumination: < 1000 lux). The culture water (so-called 'M4 medium') is partly renewed once a week. The Daphnia are exclusively fedunicellular green algae (Desmodesmus snbspicatus) 'ad libitum'. Mortalities of parent Daphnia during the culture period are recorded daily in a semi-quantitative way. The neonates are separated from their parent Daphnia by filtration prior to the reproduction test.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Post exposure observation period:

No post exposure observation period

Hardness:

The hardness of the dilution water ranged from 15.2 - 16.1 °dH (=271.32 - 287.39 mg/L CaC03) during the test.

Test temperature:

mean ± 2 °C (in the temperature range 18-22 °C)

pH:

7.9 - 8.2

Dissolved oxygen:

8.3 - 8.8 mg/l

Salinity:

Not applicable.

Nominal and measured concentrations:

Test concentration/s (nominal): 0.1, 0.316, 1.0, 3.16 and 10 mg/l

Details on test conditions:

Reconstituted water (so-called 'M4 medium', originally described in Water Research 24 (9), September 1990: 1157-1167), was prepared according to the recommendations of Bundesgesundheitsamt Berlin. This standard dilution water is used for both, the maintenance of the test animals and the preparation of stock and test solutions of the test item.

Test vessels: 150 ml glass beakers holding 1 neonate in 100 ml of test medium
Experimental design: 5 test concentrations plus 1 control. 1 neonate per vessel, 10 replicates per concentration/control. Semi-static system (renewal of the test media on Monday, Wednesday and Friday) respectively.
Method of initiation: neonates were placed in prepared media
Photoperiod: 16 h light: 8 h darkness
Temperature: mean +/- 2 °C (in the temperature range 18-22 °C)
Aeration: none
Test concentration/s (nominal): 0.1, 0.316, 1.0, 3.16 and 10 mg/l
Light intensity: not exceeding 800 lux
Food source and feeding rate: Desmodesmus subspicatus
day 0-7: 0.1 mg C / Daphnia / day
day 8-21: 0.2 mg C / Daphnia / day
Duration of exposure: 21 days
Criteria of effects: The criterion of adverse effects used in this study was the item-induced inhibitory effect on the parthenogenetic reproduction rate and the mobility behaviour/mortality rate of parent Daphnia, all effects being assessed at least three times a week.

Reference substance (positive control):

no

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

>= 10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

> 10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

>= 10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

> 10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

Based on different biological effects observed and endpoints assessed, the following results were obtained at the end of the 21 day exposure period of the Daphnia reproduction test:

Results [mg/l]

21d-NOEC (reproduction) (tα 0.05): ≥10
21d-LOEC (reproduction) (tα 0.05): >10
21d-NOEC (mortality) (tα 0.05): ≥10
21d-LOEC (mortality) (tα 0.05): >10
1st brood on day: 7

The results are expressed in terms of nominal concentrations. Recovery rates in the highest test item concentration of 10 mg/l ranged from 91.8 – 99.7% % of the nominal values in the freshly prepared media and from 85.8.0 - 97.8 % in the media after 48 hours of exposure, respectively.

COMMENTS
In the highest test concentration the reproduction exceeded 27.1% in comparison with the control. Also in the highest test concentration some of the live juveniles were lethargic and stayed on the bottom of the test vessels.

Results with reference substance (positive control):

Not applicable.

Reported statistics and error estimates:

The reproductive output of the Daphnia exposed to the test item was compared to that of the control in order to determine the Lowest Observed Effect Concentration (LOEC) and the No Observed Effect Concentration (NOEC) according to Williams Multiple Sequential t-test Procedure. Additionally, the threshold concentration with survival of the parent Daphnia was determined using Fishers Exact Binomial Test with Bonferroni Correction and the time to production of first brood recorded.

Nominal Concentration [mg/l]: Control

	Number of living offspring per replicate
	1	2	3	4	5	6	7	8	9	10
Σ (1-21)	160	171	167	142	121	141	122	191	175	185
	Number of dead offspring / aborted eggs per replicate
	1	2	3	4	5	6	7	8	9	10
Σ (1-21)	0/0	0/0	0/0	0/0	0/0	0/0	0/0	0/0	0/0	0/0

 

Nominal Concentration [mg/l]: 0.1

	Number of living offspring per replicate
	1	2	3	4	5	6	7	8	9	10
Σ (1-21)	156	181	161	175	171	162	182	197	156	150
	Number of dead offspring / aborted eggs per replicate
	1	2	3	4	5	6	7	8	9	10
Σ (1-21)	0/0	0/0	0/0	0/0	0/0	0/0	0/0	0/0	0/0	0/0

 

Nominal Concentration [mg/l]: 0.316

	Number of living offspring per replicate
	1	2	3	4	5	6	7	8	9	10
Σ (1-21)	140	157	160	171	163	160	157	180	182	163
	Number of dead offspring / aborted eggs per replicate
	1	2	3	4	5	6	7	8	9	10
Σ (1-21)	0/0	0/0	0/0	0/0	0/0	0/0	0/0	0/0	0/0	0/0

 

Nominal Concentration [mg/l]: 1.0

	Number of living offspring per replicate
	1	2	3	4	5	6	7	8	9	10
Σ (1-21)	158	187	+	171	201	141	146	166	172	160
	Number of dead offspring / aborted eggs per replicate
	1	2	3	4	5	6	7	8	9	10
Σ (1-21)	0/0	0/0	0/0	0/0	0/0	0/0	0/7	0/8	0/9	0/0

 

Nominal Concentration (mg/l]: 3.16

	Number of living offspring per replicate
	1	2	3	4	5	6	7	8	9	10
Σ (1-21)	184	+	200	157	177	177	206	201	210	225
	Number of dead offspring / aborted eggs per replicate
	1	2	3	4	5	6	7	8	9	10
Σ (1-21)	3/0	0/0	0/0	0/0	0/0	4/0	0/0	2/0	0/0	5/0

 

Nominal Concentration [mg/l]: 10

	Number of living offspring per replicate
	1	2	3	4	5	6	7	8	9	10
Σ (1-21)	185	206	208	193	215	167	197	228	+	203
	Number of dead offspring / aborted eggs per replicate
	1	2	3	4	5	6	7	8	9	10
Σ (1-21)	14/0	18/0	4/0	0/0	0/0	28/0	16/0	15/0	23/0	0/0

 

 

Validity criteria fulfilled:

yes

Conclusions:

Results [mg/l]

21d-NOEC (reproduction) (tα 0.05): ≥10
21d-LOEC (reproduction) (tα 0.05): ≥10
21d-NOEC (mortality) (tα 0.05): ≥10
21d-LOEC (mortality) (tα 0.05): 10
1st brood on day: 7

Executive summary:

A study was performed to assess the effects of Reaktiv Gelb F68072 FW on the reproductive output of parthenogenetic Daphnia magna STRAUS under semi-static conditions.

 

The study was conducted in accordance with EEC Methods for Determination of Ecotoxicity Annex to Directive 97/548/EEC Part C, Method 20 'Daphnia magnaReproduction Test" (adopted: 2001), which equals to the OECD Test guideline 211(Daphnia magnaReproduction Test; adopted: Sept. 21, 1998).This study was performed in accordance with the principles of Good Laboratory Practice (GLP) and reported with a GLP certificate.

 

Young female Daphnia, aged less than 24 hours at the start of the test, were exposed to a range of concentrations, nominally 0.1, 0.316, 1.0, 3.16 and 10 mg/l of Reaktiv Gelb F68072 FW dissolved in water.

 

 

After a 21d-exposure period, the total number of living offspring produced per parent animal alive at the end of the test was assessed. The reproductive output of the Daphnia exposed to the test item was compared to that of the control in order to determine the Lowest Observed Effect Concentration (LOEC) and the No Observed Effect Concentration (NOEC) according to Williams Multiple Sequential t-test Procedure. Additionally, the threshold concentration with survival of the parent Daphnia was determined using Fishers Exact Binomial Test with Bonferroni Correction and the time to production of first brood recorded. The results are as follows in mg/l:

 

21d-NOEC (reproduction) (tα 0.05): ≥10

21d-LOEC (reproduction) (tα 0.05): >10

21d-NOEC (mortality) (tα 0.05): ≥10

21d-LOEC (mortality) (tα 0.05): >10

1st brood on day: 7

 

The results are expressed in terms of nominal concentrations. Recovery rates in the highest test item concentration of 10 mg/l ranged from 91.8 – 99.7% % of the nominal values in the freshly prepared media and from 85.8.0 - 97.8 % in the media after 48 hours of exposure, respectively.

 

The hardness of the dilution water ranged from 15.2 - 16.1 °dH (= 271.32 - 287.39 mg/L CaCO3) during the test.

Description of key information

The NOEC for reproduction and mortality after 21d-exposure was determined to be >= 10 mg/L (nominal conc.).

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

NOEC

Remarks:

(reproduction and mortality)

Effect concentration:

>= 10 mg/L

Additional information

A study was performed to assess the effects of Reaktiv Gelb F68072 FW on the reproductive output of parthenogenetic Daphnia magna STRAUS under semi-static conditions.

 

The study was conducted in accordance with EEC Methods for Determination of Ecotoxicity Annex to Directive 97/548/EEC Part C, Method 20 'Daphnia magnaReproduction Test" (adopted: 2001), which equals to the OECD Test guideline 211(Daphnia magnaReproduction Test; adopted: Sept. 21, 1998).This study was performed in accordance with the principles of Good Laboratory Practice (GLP) and reported with a GLP certificate.

 

After a 21d-exposure period, the total number of living offspring produced per parent animal alive at the end of the test was assessed. The reproductive output of the Daphnia exposed to the test item was compared to that of the control in order to determine the Lowest Observed Effect Concentration (LOEC) and the No Observed Effect Concentration (NOEC) according to Williams Multiple Sequential t-test Procedure. Additionally, the threshold concentration with survival of the parent Daphnia was determined using Fishers Exact Binomial Test with Bonferroni Correction and the time to production of first brood recorded.

The results are expressed in terms of nominal concentrations. Recovery rates in the highest test item concentration of 10 mg/l ranged from 91.8 – 99.7% % of the nominal values in the freshly prepared media and from 85.8.0 - 97.8 % in the media after 48 hours of exposure, respectively.