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Reference
Endpoint:
activated sludge respiration inhibition testing
Type of information:
experimental study
Adequacy of study:
key study
Study period:
28 June 2017 to 28 July 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test (Carbon and Ammonium Oxidation))
GLP compliance:
yes
Analytical monitoring:
not required
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION: stock solution was prepared in deionized water (10 g/L) additional dilutions were sone with deionized water
Test organisms (species):
activated sludge of a predominantly domestic sewage
Details on inoculum:
- Name and location of sewage treatment plant where inoculum was collected: from the mixed treatment plant of urban (about 66%) and industrial (about 34%) liquid sewage situated in San Rocco near Monza (Italy)
- Method of cultivation: under aeration
- Preparation of inoculum for exposure: no settlement, sludge was washed and used on day of collection
- Initial biomass concentration: 3 g SS/L
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
3 h
Test temperature:
20°C ± 2°C
pH:
7.31-7.42 at 6.25-100 mg/L
Nominal and measured concentrations:
pre-test 1, 10 and 100 mg/L
main study: 6.25, 12.5. 25, 50 and 100 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: glass bottles
- Aeration: contimuously
- No. of vessels per concentration (replicates): 5 (3 in pre-test)
- No. of vessels per blank control (replicates): 6 (3 in pre-test)
- No. of vessels per vehicle control (replicates): 5 (3 in pre-test)
- No. of vessels per abiotic control (replicates): 0 (3 in pre-test)
- Sludge concentration (weight of dry solids per volume): 1.5 g/L
- Nutrients provided for bacteria: synthetic sewage feed (as per OECD 209)

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: deionized water
- Particulate matter: none

OTHER TEST CONDITIONS
- Adjustment of pH: yes with NaOH
- Photoperiod: NA

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : dissolved oxygen concentration over period of 6 minutes leading to calculation of the oxygen consumption rate

Reference substance (positive control):
yes
Key result
Duration:
3 h
Dose descriptor:
EC50
Effect conc.:
36.74 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
inhibition of total respiration
Details on results:
- Blank controls oxygen uptake rate: 22.28 mg oxygen per one gram of activated sludge in an hour (pretest 16.79%)
- Coefficient of variation of oxygen uptake rate in control replicates: 8.47% (pretest 0.20%)
Results with reference substance (positive control):
EC50 3,5 Dichlorophenol 6.7 mg/L
Reported statistics and error estimates:
Probit analysis

Concentration

% of inhibition total respiration rate

6.25 mg/l

1.72

12.5 mg/l

11.14

25.0 mg/l

38.70

50.0 mg/l

59.88

100.0 mg/l

86.63

 

Validity criteria fulfilled:
yes
Conclusions:
The EC50 for inhibition of respiration in bacteria from sewage sludge in 36.7 mg/L
Executive summary:

In a test according to OECD 209, bacteria in sewage sludge were exposed to 6.25, 12.5, 25, 50 and 100 mg/L of the substance during 3 hours under continuous aeration. The inhibition of total respiration was measured over a period of 6 minutes. The EC50 was 36.7 mg/L.

Description of key information

In a test according to OECD 209, bacteria in sewage sludge were exposed to 6.25, 12.5, 25, 50 and 100 mg/L of the substance during 3 hours under continuous aeration. The inhibition of total respiration was measured over a period of 6 minutes. The EC50 was 36.7 mg/L.

Key value for chemical safety assessment

EC50 for microorganisms:
36.7 mg/L

Additional information