Abstract
Fixed-film processes for wastewater treatment are becoming widely used. Their efficiency is usually estimated only from substrate removal rate measurements. A better understanding of fixed culture composition and activity is needed to optimize processes in which they are involved. These analyses often require that biofilms are removed from their substrata, but this procedure is one of the most limiting steps in biofilm investigations, especially when cell counts are involved. The main objective of the study was to develop an optimal removal procedure based on sonication for analysis of biofilm parameters such as total and active bacterial counts by epifluorescent microscopy (4',6-diamidino-2-phenylindole (DAPI) and 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) stainings) and total organic content as estimated by total proteins determination and chemical oxygen demand measurements. Experiments were carried out on nitrifying biofilms developed on a plastic granular substratum. Results confirmed that the removal step is crucial in biofilm analysis. The repeatability of bacterial enumeration was evaluated as well as the efficiency of sonication treatment and optimal conditions for attached cell removal. Within the studied range of sonication conditions, the sonication time and the duty cycle improved the removal efficiency of active bacteria, whereas the sonication power had the opposite effect.
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