In this study, a granular sludge sequencing batch reactor (GSBR) was started-up for the biological aerobic treatment of wastewater containing highly toxic 2,4-dichlorophenol (2,4-DCP), in presence of readily biodegradable sodium acetate (NaAc) as the growth substrate. Different influent concentrations of NaAc (420–800 mg/L) and 2,4-DCP (0–20 mg/L), as well as different operating conditions (i.e. cycle length), were tested in order to determine the optimal strategy for successful GSBR start-up: stable granulation and complete 2,4-DCP removal were achieved only when high NaAc influent concentration and volumetric organic loading rates (800 mg/L and 1.9 kgCOD/(m3·d), respectively), prolonged reaction phase (cycle time of 4 hours) and gradual increase of 2,4-DCP concentration in the influent were applied, thus providing useful information for process optimization in view of future scale-up. Granules were initially colonized by fungi which progressively disappeared during the start-up process, and complete 2,4-DCP removal was mostly due to bacterial activity, in particular Betaproteobacteria, as shown by fluorescence in situ hybridization (FISH).

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