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Ecotoxicological information

Toxicity to microorganisms

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Reference
Endpoint:
toxicity to microorganisms, other
Type of information:
experimental study
Adequacy of study:
key study
Study period:
December 2006
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
No CoA included in report, but molecular formula & weight were provided by the study sponsor. Not all validity criteria discussed and volatility of test substance not shown/given to conclude on test system suitability.
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
other: OECD 301F (manometric respiratory test)
Qualifier:
according to guideline
Guideline:
other: EU Method C.4D
GLP compliance:
yes (incl. QA statement)
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The test item was weighed by means of an analytical balance and transferred to the test flasks with water. No emulsifiers or solvents were used.
- Controls: In this study the test item was tested and in parallel an inoculum control, procedure control (with reference substance), abiotic control and toxicity control (with the test substance and reference substance)
Test organisms (species):
activated sludge of a predominantly domestic sewage
Details on inoculum:
- Source of inoculum/activated sludge: Aerobic activated sludge from a waste water treatment plant ARA Ergolz II, Fullinsdorf, Switzerland treating predominantly domestic sewage.
- Preparation of inoculum for exposure: The sludge was washed twice with tap water by centrifugation and the supernatant liquid phase was decanted. A homogenized aliquot of the final sludge suspension was weighed thereafter dried and the ration of wet to dry weight was calculated. Calculated amounts of wet sludge were suspended in the test water to obtain a concentration equivalent to 4 g (10%) dry material per liter. During the holding, the sludge was aered at room temperature until use. Prior to use, the sludge was aerated at room temperature until use. Prior to use, the sludge was first thoroughly mixed and then diluted with test water at a concentration of 1 g per liter (dry weight basis).
- Concentration of sludge: Based on the determined dry weight of this diluted activated sludge defined amounts were added to test water to obtain a final concetration of 30 mg dry material per liter.
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
28 d
Test temperature:
22 °C
pH:
t= 0 7.4
t = 28 d 7.4- 7.7
Nominal and measured concentrations:
Nominal: 101 mg/L test substance plus 100 mg/L reference substance sodium benzoate
Details on test conditions:
TEST CONDITIONS
- Composition of medium: Test water was prepared according to the test guidelines OECD 301F.
- pH adjusted: yes, to obtain the final test water the pH was adjusted from 7.8 to 7.4 with diluted hrdochloric acid solution.
- Aeration of dilution water: The sludge was aered at room temperature prior to use. Aeration of dilution water not specified.
- Continuous darkness: yes

TEST SYSTEM
- Culturing apparatus: 500 mL Erlenmeyer flasks. The test flasks were incubated under continuous stirring in a SAPROMAT D12 (voith GmbH, Heidenheim, Germany).
- Number of culture flasks/concentration: 2
- Measuring equipment: Electrode type manometer
- Details of trap for CO2 and volatile organics if used: The CO2 is absorbed by soda lime and the total pressure drop is detected and converted into an electrical signal by means of an electrode type manometer. The consumed oxygen is replaced by electrolytically generated oxygen froma copper sulfate solution.

SAMPLING
- Sampling frequency: Oxygen consumption was recorded manually by taking a daily reading at least on each working day.
- Sampling method: manually

CONTROL AND BLANK SYSTEM
- Inoculum blank: yes
- Abiotic sterile control: yes
- Toxicity control: yes, with reference item sodium benzoate

Reference substance (positive control):
no
Key result
Duration:
14 d
Dose descriptor:
NOEC
Effect conc.:
101 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
inhibition of total respiration
Details on results:
- Main result regarding toxicity to microorganisms: Biodegradation in the toxicity control: The percent biodegradation in the toxicity control, containing both the test item and the reference item, was calculted based on the sum of the ThoD of the test item and the reference item. In the toxicity control, the course of biodegradation over the 28-day exposure period was similar to the two procedure controls, containing only the reference item. Within 14 days of exposure, biodegradation amounted to 35% based on the ThoD.
Thus according to the test guidelines (OECD301F), the test item had no inhibitory effect on activated sludge microorganisms at the tested concentration of 100 mg/L because biodegradation in the toxicity control was > 25% within 14 days.

- Additional results (OECD 301F study):
Biodegradation of the reference item: The percent biodegradation of the reference item sodium benzoate was calculated based on the theoretical oxygen demand of 1.67 O2/mg. In the procedure controls, the reference item was degraded by an average of 88% by exposure day 14. At the end of the test (day 28), the reference item was degraded by an average of 91%.

Abiotic control: No degradation of the test item occured in the abiotic control under the test conditions within 28 days.

The biochemical oxygen demand of the test item in the test media significantly increased from exposure day 5 until test termination after 28 days. At the end of the 28-day exposure period, the mean biodegradation of the test item amounted to 55%. Consequently the test item was found to be biodegradable under the test conditions within 28 days. However the pass level for ready biodegradabiliy, i.e. biodegradation of at least 60% of the ThoD in a 10-day window within the 28-day period of the test, was not reached.


Validity criteria fulfilled:
yes
Remarks:
based on criteria for the OECD TG301F
Conclusions:
Under the conditions of the test, 101 mg test substance did not cause toxicity to activated sludge microorganisms.
Executive summary:

A manometric respiratory test was performed according to OECD TG 301F and in compliance with GLP. The oxygen consumption was followed for 28-days in an aerobic system inoculated with domestic activated sludge. 101.0 mg/L test substance (274 mg/L ThOD) was added to the test system. The experiment was performed in duplicate and inoculum control, procedure control, abiotic control, and toxicity control were performed in parallel. The toxicity control was performed with the test substance and reference substance sodium benzoate (100 mg/L). In the procedure controls, the reference item was degraded by an average of 88% by exposure day 14. At the end of the test (day 28), the reference item was degraded by an average of 91%. The percent biodegradation in the toxicity control, containing both the test item and the reference item, was calculated based on the sum of the ThOD of the test item and the reference item. In the toxicity control, the course of biodegradation over the 28-day exposure period was similar to the two procedure controls, containing only the reference item. Within 14 days of exposure, biodegradation amounted to 35% based on the ThOD. Thus according to the test guidelines (OECD301F), the test item had no inhibitory effect on activated sludge microorganisms at the tested concentration of 100 mg/L because biodegradation in the toxicity control was > 25% within 14 days. Under the conditions of the test, 101 mg test substance did not cause toxicity to activated sludge microorganisms and hence was considered the NOEC.

Description of key information

A manometric respiratory test was performed according to OECD TG 301F and in compliance with GLP. The oxygen consumption was followed for 28-days in an aerobic system inoculated with domestic activated sludge. 101.0 mg/L test substance (274 mg/L ThOD) was added to the test system. The experiment was performed in duplicate and inoculum control, procedure control, abiotic control, and toxicity control were performed in parallel. The toxicity control was performed with the test substance and reference substance sodium benzoate (100 mg/L). In the procedure controls, the reference item was degraded by an average of 88% by exposure day 14. At the end of the test (day 28), the reference item was degraded by an average of 91%. The percent biodegradation in the toxicity control, containing both the test item and the reference item, was calculated based on the sum of the ThOD of the test item and the reference item. In the toxicity control, the course of biodegradation over the 28-day exposure period was similar to the two procedure controls, containing only the reference item. Within 14 days of exposure, biodegradation amounted to 35% based on the ThOD. Thus according to the test guidelines (OECD301F), the test item had no inhibitory effect on activated sludge microorganisms at the tested concentration of 100 mg/L because biodegradation in the toxicity control was > 25% within 14 days. Under the conditions of the test, 100 mg test substance did not cause toxicity to activated sludge microorganisms and hence was considered the NOEC.

Key value for chemical safety assessment

EC10 or NOEC for microorganisms:
100 mg/L

Additional information