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Toxicity to microorganisms

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Reference
Endpoint:
activated sludge respiration inhibition testing
Type of information:
experimental study
Adequacy of study:
key study
Study period:
09 February 2012 - 09 February 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.11 (Biodegradation: Activated Sludge Respiration Inhibition Test)
Deviations:
no
Qualifier:
according to
Guideline:
ISO 8192 (Water quality - Test for inhibition of oxygen consumption by activated sludge for carbonaceous and ammonium oxidation)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
Silsequioxanes, phenyl (tested as received) was a clear colourless liquid that was not completely soluble in test medium at the loading rates initially prepared. Therefore, 1-Litre test bottles were filled with 200 ml of test substance mixtures in Milli-RO water (tap water purified by reverse osmosis; Millipore Corp., Bedford, Mass., USA) with initial loading rates of 2.5 times the final loading rate. These mixtures were stirred in closed dark brown bottles for 23½ hours, except in the abiotic control (25½ hours). Subsequently, 16 ml synthetic sewage feed, 250 ml sludge and Milli-RO water up to 500 ml were added resulting in the required loading rates. Optimal contact between the test substance and test organisms was ensured applying continuous aeration and stirring.
Test organisms (species):
activated sludge of a predominantly domestic sewage
Details on inoculum:
The sludge was coarsely sieved, washed and diluted with ISO-medium. A small amount of the sludge was weighed and dried overnight at ca. 105°C to determine the amount of suspended solids (3.0 g/l of sludge, as used for the test). The pH was 7.4 on the day of testing. The batch of sludge was used one day after collection; therefore 50 ml of synthetic sewage feed was added per litre of activated sludge at the end of the collection day. The sludge was kept aerated at test temperature until use.
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
3 h
Post exposure observation period:
After the 3-hour contact time, the oxygen consumption was recorded for a period of approximately 10 minutes. During measurement, the sample was not aerated but continuously stirred on a magnetic stirrer. The pH and temperature were determined in the remaining part of the reaction mixture.
Test temperature:
Between 18.1 and 20.5°C.
pH:
Before addition of sludge : 7.6 - 7.7.
After the 3 hour exposure period : 7.5 - 8.3.
Dissolved oxygen:
The oxygen concentration at the start of measurements was at least 7.0 mg O2/l

Oxygen consumption:
Controls : 48 mg O2/l/h (mean value)
silsequioxanes, phenyl 10 mg/l : 42 mg O2/l/h
silsequioxanes, phenyl 100 mg/l : 49 mg O2/l/h
silsequioxanes, phenyl 1000 mg/l : 51 mg O2/l/h (mean value)
Nominal and measured concentrations:
Nominal loading rates: 10, 100 and 1000 mg/l.
Details on test conditions:
TEST SYSTEM
- Type: open
- Material, size, headspace, fill volume: All glass
- Aeration: During exposure with clean, oil-free air
- No. of vessels per concentration (replicates): 1 for 10 and 100 mg/l and 3 for 1000 mg/l
- No. of vessels per control (replicates): 2
- Biomass loading rate: 1.5 g/l suspended solids in final test mixture

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Tap-water purified by reverse osmosis (Milli-RO
EFFECT PARAMETERS MEASURED (with observation intervals if applicable): The respiration rate from each vessel, in mg O2/l/hr, was calculated from the linear part of the respiration curve, which was generally between 2 and 7 mg O2/l.

TEST CONCENTRATIONS
- Test concentrations: combined limit/range-finding test: at 10, 100 and 1000 mg/l (loading rates).
Reference substance (positive control):
yes
Remarks:
3,5-dichlorophenol
Duration:
3 h
Dose descriptor:
NOEC
Effect conc.:
>= 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
(loading rate)
Basis for effect:
inhibition of total respiration
Remarks:
respiration rate
Duration:
3 h
Dose descriptor:
EC50
Effect conc.:
> 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
(loading rate)
Basis for effect:
inhibition of total respiration
Remarks:
respiration rate
Details on results:
In the combined limit/range-finding test no inhibition of the respiration rate of the sludge was recorded at the highest loading rate tested, 1000 mg silsequioxanes, phenyl per litre.

There was no oxygen uptake from abiotic processes and the results at 1000 mg/l with a nitrification inhibitor showed that the heterotrophic inhibition of the respiration rate was comparable to the total inhibition.
Results with reference substance (positive control):
The EC50 of 3,5-dichlorophenol was in the accepted range of 2 to 25 mg/l for total respiration (4.0 mg/l).
Reported statistics and error estimates:
ECx
For the reference substance the percentage inhibition was plotted against the logarithm of the concentrations and the EC50 was determined using linear regression analysis.

For silsequioxanes, phenyl no EC50 could be calculated because the test substance proved to be non-toxic (EC50 > a loading rate of 1000 mg/l).

NOEC estimation
Since no inhibition of the respiration rate was observed at the highest loading rate tested, no statistical analysis of the data was performed.
Validity criteria fulfilled:
yes
Conclusions:
Under the conditions of this present test silsequioxanes, phenyl was not toxic to waste water (activated sludge) bacteria up to and including a loading rate of 1000 mg/l (NOEC).

The EC50 exceeded a loading rate of 1000 mg/l.

Description of key information

Toxicity to microorganisms: 3 hour EC50 >1000 mg/l; NOEC >=1000 mg/l (OECD 209), read-across from phenyl silsesquioxanes ( EC 939-487-8).

Key value for chemical safety assessment

Additional information

There are no microorganism toxicity data available for 3,3-bis[(dimethylvinylsilyl)oxy]-1,1,5,5-tetramethyl-1,5-divinyltrisiloxane (ViM4Q).

A category approach is applied to this endpoint and is detailed in the Siloxane Category report (PFA, 2017a). The hypothesis for read-across is that siloxane substances exhibit no evidence of significant toxicity to sewage treatment plant microorganisms. This is consistent with a non-polar narcosis mode of action. Therefore, good quality data for the structurally-related substance phenyl silsesquioxanes (EC 939-487-8), have been read across to ViM4Q.

ViM4Q and phenyl silsesquioxanes are members of the Reconsile Siloxanes Category within which, in general, the substances exhibit no evidence of significant toxicity to microorganisms.

The table below presents microorganism toxicity data available for substances within the Siloxanes Category.

It is therefore considered valid to read-across the results for phenyl silsesquioxanes to fill the data gap for the registered substance.


Microorganism toxicity data available for substances within the Siloxanes Category

CAS

Name

Result: E(I)C50(mg/l)

Result: NOEC (or EC10/ EC20) (mg/l)

Guideline Number

Test method

Duration (h)

Reliability

540-97-6

Dodecamethylcyclohexasiloxane

>100 mg/l

 

OECD 209

ASRI

3

1

541-05-9

Hexamethylcyclotrisiloxane

>100 mg/l

>100 mg/l

OECD 209

ASRI

3

1

541-02-6

Decamethylcyclopentasiloxane

>2000 mg/l

 

C11

ASRI

3

1

107-46-0

Hexamethyldisiloxane

>>100

≥100

OECD 209

ASRI

3

1

141-62-8

Decamethyltetrasiloxane

>100 mg/l

 

OECD 209

ASRI

3

1

352230-22-9

Siloxanes and Silicones, di-Ph, Ph (trimethylsilyl)oxy

>1000 mg/l

≥1000 mg/l

OECD 209

ASRI

3

1

2627-95-4

1,1,3,3-tetramethyl-1,3-divinyldisiloxane

>1000 mg/l

≥1000 mg/l

OECD 209

ASRI

3

1

2554-06-5

2,4,6,8-tetramethyl-2,4,6,8-tetravinylcyclotetrasiloxane

>1000 mg/l

≥1000 mg/l

OECD 209

ASRI

3

1

N/A (EC 939-487-8)

phenyl silsesquioxanes

>1000

≥1000

OECD 209

ASRI

3

1

99363-37-8

dimethyl, methyl(3-aminopropyl) siloxane, trimethylsiloxy-terminated

> water soluble fraction

≥ water soluble fraction

OECD 209

ASRI

3

1

71750-79-3

dimethyl, (aminoethylaminopropyl)methyl siloxane, trimethylsiloxy-terminated

> water soluble fraction

≥ water soluble fraction

OECD 209

ASRI

3

1


Additional information is given in a supporting report (PFA, 2017a) attached in Section 13 of the IUCLID dossier.

Under the conditions of this present test, phenyl silsesquioxanes was not toxic to waste water (activated sludge) bacteria up to and including a loading rate of 1000 mg/l (NOEC).