Nitrogen (N) removal from high-strength wastewater can be accomplished in single-stage combined nitritation-anammox reactors with suspended growth biomass composed of floccular sludge, granular sludge, or of any mix of these 2 different sludge fractions. To date, the influence of floccular biomass on granular sludge reactor performance and stability has not been investigated experimentally or numerically. To address this knowledge gap, two 1D multi-species models were developed in Aquasim to assess the importance of small levels of flocs in putatively granular sludge combined nitritation-anammox reactors for different bulk oxygen concentrations and organics loads. The models included the growth and decay of aerobic ammonium-oxidizing organism (AOO), nitrite-oxidizing organisms (NOO), heterotrophic organisms (OHO), and anammox organisms (AMO) in exclusively granular sludge reactors, and in granular sludge reactors with small levels (∼5% of total biomass) of flocs. While maximum N removal efficiencies were similar for both model structures, floc addition led to a lower optimal dissolved oxygen concentration (DO) as well as a narrower maximum N removal peak, suggesting that small levels of floccular material may decrease process robustness to bulk oxygen changes. For some DO levels, this led to drastic efficiency drops. Furthermore, floc addition also led to substantial segregation in activity and microbial population distribution, with AOO, NOO and OHO concentrated in flocs and AMO concentrated in granules. Increased organic loading (COD:N = 4:3) improved maximum N removal efficiency in both model structures, but yielded substantially different predictions for optimal DO setpoint and process robustness to variations in DO. Taken together, our results indicate that even small levels of floccular biomass in biofilm reactors can have profound implications for reactor performance and optimization and for segregation of linked microbial processes, and suggest that the common practice of neglecting small levels of floccular material in biofilm models and in practice may lead to erroneous predictions.

A model describing a given system should be as simple as possible – but not simpler. The appropriate level of complexity depends both on the type of system and on the intended use of the model. This paper addresses the critical question of which purposes justify increased complexity of biofilm (reactor) models. Additional model features compared to conventional models considered are: (1) the inclusion of microbial diversity, distinguishing between different species performing the same function; and (2) the distinction between flocs and granules in putatively granular sludge reactors. With a multispecies model considering interspecies diversity, it was demonstrated that a given macroscopic reactor performance does not necessarily reflect steady state conditions on the microscale. In a second case study, it was shown that the addition of a small level of flocs can have a significant impact on macroscale process performance and on microbial population and activity distributions in granular sludge reactors. It was concluded that increased complexity in biofilm models, concerning microbial diversity or mesoscale aggregate architecture, is likely more useful when the focus is on understanding fundamental microscale outputs, but under specific conditions, these additional model features can be critically informative for bulk reactor behavior prediction and general understanding.