Aerobic granular sludge (AGS) has been considered a breakthrough within the wastewater treatment sector. However, the long reactor start-up for the formation of granules is challenging and may hinder the spread of this technology. To circumvent this obstacle, inoculation of the reactors with pre-formed granules from existing plants is an interesting approach. In this context, issues related to biomass storage becomes very relevant. In this study, reactivation of aerobic granular biomass after storage was evaluated in a sequencing batch reactor (SBR) designed for achieving simultaneous organic matter, nitrogen and phosphorus removal. Two different scenarios, short (40 days) and long (180 days) storage periods, were assessed, and their influence on the granules physical properties and bioactivity was addressed. The results revealed that the granules stored for a shorter period showed higher resistance to breakage and underwent smooth color changes. On the other hand, the biomass stored for a longer period acquired a dark color and was more susceptible to disruption during reactivation. The granules stored for 6 months become swollen and exhibited an irregular morphology and fluffy structure within the first days of reactivation. Consequently, their settling properties were adversely affected, and some parameters such as the food-to-microorganism ratio had to be adjusted to prevent granules disintegration. Regarding the bioactivity of important microbial functional groups, COD removal was rapidly restored within a few days of SBR operation with the biomass stored for a shorter period. However, it took longer for the biomass stored for 180 days to reach the same performance observed for the granules stored for 40 days. A similar trend is valid for nitrification. In the experiments with sludge stored for a longer time, it took almost twice as long to reach effluent ammonium concentrations lower than 1 mg NH4+-N L−1 compared to the test using biomass stored for 40 days. Phosphate removal was strongly affected by biomass storage, especially after 180 days of inactivity, a condition found to be detrimental for polyphosphate-accumulating organisms. Finally, cycle tests were also conducted to assess substrate conversion rates for comparison between different trials and evaluate the influence of temperature (10–35 °C) on nitrification and phosphate removal rates.
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