Calcium 4-chloro-2-(5-hydroxy-3-methyl-1-(3-sulfonatophenyl)pyrazol-4-ylazo)-5-methylbenzenesulfonate

Toxicity to soil macroorganisms except arthropods

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

toxicity to soil macroorganisms except arthropods: long-term

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

According to OECD 207 and GLP

Qualifier:

according to guideline

Guideline:

OECD Guideline 207 (Earthworm, Acute Toxicity Tests)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

no

Details on sampling:

No analytical monitoring

Vehicle:

yes

Details on preparation and application of test substrate:

- Method of mixing into soil (if used): The test item was weighed out on quartz sand (5 g/replicate).
- Method of mixing into soil (if used): The weighed test item with quartz sand given to the artificial soil. Demineralised water was added to adjust 60 % of the maximum water capacity of the artifical soil. Thereafter the test medium was mixed with a long handled blender for at least 3 minutes and filled into the test vessels.
- Controls: Artificial soil without test item and 5 g quartz sand was used as control medium.

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

TEST ORGANISM
- Common name: Eisenia fetida Savigny 1826 (Annelida, Lumbricidae)
- Subspecies: Eisenia fetida andrei
- Source: Bundesanstalt fuer Verbraucherschutz und Landwirtschaft (BVL), Messeweg 11-12, D-38104 Braunschweig
- Age at test initiation (mean and range, SD): adult (with clitellum)
- Weight at test initiation (mean and range, SD): 0.45 - 0.60 g/worm


ACCLIMATION
- Acclimation period: 2 days
- Acclimation conditions (same as test or not):same as test
- Health during acclimation (any mortality observed): No mortality or abnormal behaviour observed

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

yes

Total exposure duration:

14 d

Test temperature:

17-21 °C

pH:

pH-values

. day 0 day 14
control 5.76 5.79
1000 mg/kg dw 5.78 5.64

Moisture:

Moisture [%] of dry weight

. day 0 day 14
control 48.6 51.6
1000 mg/kg dw 48.2 47.4

Details on test conditions:

TEST SYSTEM
- Test container (material, size): Glass dishes (volume 1.5 L), covered with perforated plastic film to prevent the test medium from drying up
- No. of organisms per container (treatment): 10 per container (40 per treatment)
- No. of replicates per treatment group: 4
- No. of replicates per control: 4
- No. of replicates per vehicle control: 4


SOURCE AND PROPERTIES OF SUBSTRATE (if soil)

- Composition (if artificial substrate): 10 % peat, 20 % kaolin, 69 % quartz sand, 0.37 % calcium carbonate
- Organic carbon (%): Not measured
- Maximum water holding capacity (in % dry weigth): 81
- Pretreatment of soil: demineralised water was added to give an overall moisture content of about 30 % of the maximum water holding capacitiy of the soil.
- Storage (condition, duration): room temperature, protected from light, 16 days



OTHER TEST CONDITIONS
- Photoperiod: 16
- Light intensity: 400 - 800 lx


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
The live weight of the earthworms was measured individually at the day of application (day 0) and at day 14.
Mortality, behaviour and morphological changes of the earthworms were recorded 7 and 14 days after application.

Observation parameter
a) no obvious pathological symptoms
b) no reaction to touching
c) no negative phototactical reaction
d) spasmodic winding
e) spontaneous segmentation and separation
f) yellow excretion from the oral aperture
g) ulcer and skin bleedings
t) dead earthworms


VEHICLE CONTROL PERFORMED: Yes


TEST CONCENTRATIONS

- Range finding study: a preliminary non-GLP range finding test was performed with concentrations 1 - 10 - 100 - 1000 mg/kg DW. Now effects were observed during the test duration. Therefore, the definitive study was conducted as limit test with the concentration of 1000 mg/kg DW.

Nominal and measured concentrations:

Nominal concentration: 1000 mg/kg soil dw

Reference substance (positive control):

yes

Remarks:

2-Chloroacetamide

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: mortality, loss of body weight, other symptoms

Details on results:

- Mortality at end of exposure period: No mortality observed in the control and in the test item concentration
- Morphological abnormalities: No
- Behavioural abnormalities: No
- Other biological observations: No

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- Relevant effect levels: LC50: 27.8 (CI: 26.4 - 29.2) mg/kg soil DW, NOEC: 15.0 mg/kg soil DW

Reported statistics and error estimates:

One Way Analysis of Variance (ANOVA) was carried out for the determination of statistically significant effects in comparison with control replicates. A Normality Test and an Equal Equal Variance Test were conducted prior to ANOVA. P-values for both Normality and Equal Variance Test were < 0.05. The α-value for ANOVA (acceptable probability of incorrectly concluding that there is no difference) is α = 0.05.

No mortality occurred at the limit test item concentration of 1000 mg/kg soil dry weight as well as in the controls after 14 days of exposure to the test item.

During the test the biomass loss of the earthworms was below 20 % in the controls. After 14 days of exposure the earthworm biomass was not statistically significant different at the limit test item concentration of 1000 mg/kg soil dry weight compared to the pooled control.

Mean loss of body weight

	mean loss of body weight
nominal concentration [mg/kg DW]	test start [g/worm]	test end [g/worm]	loss of body weight [%]
control	0.53	0.55	- 4
1000	0.53	0.56	- 6

Mean biomass body weight on day 14 compared to control group, 4 replicates (n = 40)

nominal concentration [mg/kg DW]	mean living weight [g] replicate
	1	2	3	4	MV ± SD	coefficient of variation	statistically significant
control	0.56	0.51	0.57	0.54	0.55 ± 0.06	11	-
1000	0.54	0.58	0.57	0.57	0.56 ± 0.05	9	no

Validity criteria fulfilled:

yes

Conclusions:

No mortality as well as no pathological symptoms or changes in the earthworm behaviour were observed at the limit test item concentration of 1000 mg/kg soil dry weight after 14 days of exposure to Pigment Yellow 191. Compared to the control the biomass of earthworms exposed to test item at the limit test item concentration of 1000 mg/kg soil dry weight was not statistically significantly reduced at test end.
Consequently, the No Observed Effect Concentration (NOEC) of the test item with respect to mortality, loss of body weight and other symptoms was determined to be at 1000 mg/kg soil dry weight.

Executive summary:

The acute effects of the test item on the earthworm Eisenia fetida were determined according to the OECD Guideline 207. The study was conducted with a limit test item concentration of 1000 mg/kg soil dry weight throughout 14 days. Artificial soil without test item was used as control. 40 test organisms with an individual weight between 0.45 - 0.6 g, divided into four replicates, were tested at the limit test item concentration and controls in artificial soil containing 10 % peat.

No mortality as well as no pathological symptoms or changes in the earthworm behaviour were observed at the limit test item concentration of 1000 mg/kg soil dry weight after 14 days of exposure. Compared to the pooled control, the biomass of the earthworms at the limit test item concentration of 1000 mg/kg soil dry weight was not statistically significantly reduced at test end.

Consequently, the no observed effect concentration (NOEC) with respect to mortality, loss of biomass and other symptoms was determined to be at 1000 mg/kg soil dry weight.

All validity criteria of the test guideline were fulfilled.

Reference 2

Endpoint:

toxicity to soil macroorganisms except arthropods: short-term

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reason / purpose for cross-reference:

read-across source

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: mortality, loss of body weight, other symptoms

Details on results:

- Mortality at end of exposure period: No mortality observed in the control and in the test item concentration
- Morphological abnormalities: No
- Behavioural abnormalities: No
- Other biological observations: No

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- Relevant effect levels: LC50: 27.8 (CI: 26.4 - 29.2) mg/kg soil DW, NOEC: 15.0 mg/kg soil DW

Reported statistics and error estimates:

One Way Analysis of Variance (ANOVA) was carried out for the determination of statistically significant effects in comparison with control replicates. A Normality Test and an Equal Equal Variance Test were conducted prior to ANOVA. P-values for both Normality and Equal Variance Test were < 0.05. The α-value for ANOVA (acceptable probability of incorrectly concluding that there is no difference) is α = 0.05.

No mortality occurred at the limit test item concentration of 1000 mg/kg soil dry weight as well as in the controls after 14 days of exposure to the test item.

During the test the biomass loss of the earthworms was below 20 % in the controls. After 14 days of exposure the earthworm biomass was not statistically significant different at the limit test item concentration of 1000 mg/kg soil dry weight compared to the pooled control.

Mean loss of body weight

	mean loss of body weight
nominal concentration [mg/kg DW]	test start [g/worm]	test end [g/worm]	loss of body weight [%]
control	0.53	0.55	- 4
1000	0.53	0.56	- 6

Mean biomass body weight on day 14 compared to control group, 4 replicates (n = 40)

nominal concentration [mg/kg DW]	mean living weight [g] replicate
	1	2	3	4	MV ± SD	coefficient of variation	statistically significant
control	0.56	0.51	0.57	0.54	0.55 ± 0.06	11	-
1000	0.54	0.58	0.57	0.57	0.56 ± 0.05	9	no

Validity criteria fulfilled:

yes

Conclusions:

No mortality as well as no pathological symptoms or changes in the earthworm behaviour were observed at the limit test item concentration of 1000 mg/kg soil dry weight after 14 days of exposure to Pigment Yellow 191. Compared to the control the biomass of earthworms exposed to test item at the limit test item concentration of 1000 mg/kg soil dry weight was not statistically significantly reduced at test end.
Consequently, the No Observed Effect Concentration (NOEC) of the test item with respect to mortality, loss of body weight and other symptoms was determined to be at 1000 mg/kg soil dry weight.

Executive summary:

The acute effects of the test item on the earthworm Eisenia fetida were determined according to the OECD Guideline 207. The study was conducted with a limit test item concentration of 1000 mg/kg soil dry weight throughout 14 days. Artificial soil without test item was used as control. 40 test organisms with an individual weight between 0.45 - 0.6 g, divided into four replicates, were tested at the limit test item concentration and controls in artificial soil containing 10 % peat.

No mortality as well as no pathological symptoms or changes in the earthworm behaviour were observed at the limit test item concentration of 1000 mg/kg soil dry weight after 14 days of exposure. Compared to the pooled control, the biomass of the earthworms at the limit test item concentration of 1000 mg/kg soil dry weight was not statistically significantly reduced at test end.

Consequently, the no observed effect concentration (NOEC) with respect to mortality, loss of biomass and other symptoms was determined to be at 1000 mg/kg soil dry weight.

All validity criteria of the test guideline were fulfilled.

Description of key information

The acute effects of the test item on the earthworm Eisenia fetida were determined according to the OECD Guideline 207. The study was conducted with a limit test item concentration of 1000 mg/kg soil dry weight throughout 14 days.

No mortality as well as no noteworthy pathological symptoms or changes in the earthworm behaviour were observed at the limit test item concentrationof 1000 mg/kg soil dry weight after 14 days of exposure. Compared to the pooled control, the biomass of the earthworms at the limit test item concentration of 1000 mg/kg soil dry weight was not statistically significantly reducedat test end.

Consequently, the no observed effect concentration (NOEC) with respect to mortality, loss of biomass and other symptoms was determined to be at 1000 mg/kg soil dry weight.