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A 1 L Erlenmeyer flask was equipped with an oxygen electrode connected to a recorder. The solution for the SOUR test for ammonium oxidizers composed of 100 mg N/L ammonium and other minerals without any organic compound or nitrite. The solution for nitrite oxidizers composed of 100 mg N/L nitrite and other minerals without organic compound or ammonium. A control test contained only minerals. A mixed liquor sample or a detached-biofilm suspension was saturated with oxygen by shaking and transferred to the flask.The initial biomass concentration in the batch reactors was in the range of 700¨C1000 mg VSS/L for suspended growth cells and 150¨C300 mg VSS/L for detached biofilm cells. The flask was carefully closed without leaving air bubbles. For the attached cell experiment, the detachment of the biofilm from the surface of the media collected from the reactor was performed using a vortex (MO Bio Vortex) for 5 min at the maximum speed and by adding polyox-yethylene lauryl ether to aid in removing cells from surfaces and promoting
their lyses . This procedure was repeated three times. The degree of detachment was monitored to determine the suitability of the method by measuring microbial activity remaining in the medium by adenosine triphosphate (ATP) and no activity was observed in the medium (data not shown).The cell suspension was stirred with a magnetic stirrer at 150 rpm, 30 8C and pH 8, respectively. The SOUR (mg O
2/g VSS d) of each sample was equal to the slope of DO depletion versus time divided by the VSS in the flask.
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