Microbial biofilms are ubiquitous in aquatic environments where they provide important ecosystem functions. A key property believed to influence the community structure and function of biofilms is thickness. However, since biofilm thickness is inextricably linked to external factors such as water flow, temperature, development age and nutrient conditions, its importance is difficult to quantify. Here, we designed an experimental system in a wastewater treatment plant whereby nitrifying biofilms with different thicknesses (50 or 400 µm) were grown in a single reactor, and thus subjected to identical external conditions. The 50 and 400 µm biofilm communities were significantly different. This beta-diversity between biofilms of different thickness was primarily caused by deterministic factors. Turnover (species replacement) contributed more than nestedness (species loss) to the beta-diversity, i.e. the 50 µm communities were not simply a subset of the 400 µm communities. Moreover, the two communities differed in the composition of nitrogen-transforming bacteria and in nitrogen transformation rates. The study illustrates that biofilm thickness alone is a key driver for community composition and ecosystem function, which has implications for biotechnological applications and for our general understanding of biofilm ecology.

This study evaluates the effect of biofilm thickness on the nitrifying activity in moving bed biofilm reactors (MBBRs) in a controlled environment. In-depth understanding of biofilm properties in MBBRs and their effect on the overall treatment efficiency is the key to optimizing process stability and efficiency. However, evaluating biofilm properties in continuously operated MBBRs can be extremely challenging. This study uses a carrier design which enables comparison of four different biofilm thicknesses, in otherwise equally operated lab-scale MBBRs. The results show that within the studied range (200-500 µm) and specific operation conditions, biofilm thickness alone had no significant effect on the overall ammonium removal. The nitrate production, however, decreased with a decreasing biofilm thickness, and the ratio between nitrite and ammonia-oxidizing activity decreased both with increasing load and decreasing oxygen concentration for all thicknesses. The suggestion that nitratation is disfavoured in thin biofilms is an interesting contribution to the current research being performed on nitrite-oxidizing bacteria inhibition for deammonification applications. By indicating that different groups of bacteria respond differently to biofilm thickness, this study accentuates the importance of further evaluation of these complex systems.

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A new principle for mainstream nitrogen removal through nitritation followed by anammox was studied in a two-stage moving bed biofilm reactor (MBBR) configuration. The first stage was optimized for nitritation by using thin biofilms and a feed alternating between synthetic mainstream wastewater at 15°C and, for shorter periods, synthetic reject water at 30 °C. The exposure of the biofilm to reject water conditions aimed to improve the growth conditions for ammonia oxidizing bacteria, while inhibiting nitrite oxidizing bacteria. The biofilm thickness was maintained below 200 μm to ensure high exposure of the total biomass to the bulk reactor conditions. Nitritation was successfully achieved in the configuration, with a nitrite accumulation ratio above 75% during the majority of the study, and ammonia removal rates between 0.25 and 0.50 g NH4-N/L,d. The anoxic second stage, optimized for anammox, was fed with the effluent from the nitritation reactor, reaching nitrogen removal rates above 0.20 g TN/L,d.

The moving bed biofilm reactor (MBBR) technology is a proven standalone and add-on technology for carbon and nutrient removal from municipal wastewaters. The key challenge of the carbon removal MBBR...