Spatiotemporal succession of phosphorous accumulating biofilms during the first year of establishment.

Many wastewater treatment plants are dependent on the utilization of microorganisms in biofilms. Our knowledge about the establishment of these biofilms is limited, particular with respect to biofilms involved in enhanced biological phosphorus removal (EBPR). These biofilms rely on polyphosphate-accumulating organisms (PAOs), requiring alternating oxic and anaerobic conditions for phosphorous uptake. This challenge has been solved using the Hias process, which combines moving-bed biofilm-reactor (MBBR) technology with physical transfer of biofilm-carriers from oxic to anaerobic zones. We combined biofilm fractionation with temporal analyses to unveil the establishment in the Hias process. A stable phosphorous removal efficiency of >95% was reached within 16 weeks of operation. Phosphorus removal, however, was not correlated with the establishment of known PAOs. The biofilms seemed associated with an outer microbiota layer with rapid turnover and an inner layer with a slow expansion. The inner layer showed an overrepresentation of known PAOs. In conclusion, our spatiotemporal analyses of phosphorous accumulating biofilm establishment lead to a new model for biofilm growth, while the mechanisms for phosphorous removal remain largely unresolved.

Spatial fractionation of phosphorus accumulating biofilm: stratification of polyphosphate accumulation and dissimilatory nitrogen metabolism.

The spatial distribution of microorganisms represents a critical issue in understanding biofilm function. The aim of the current work was to develop a method for biofilm fractionation, facilitating the analysis of individual spatial biofilm layers using metagenomic approaches. Phosphorus accumulating biofilm applied in an enhanced biological phosphorus removal wastewater treatment plant, were fractionated, and analyzed. The fractionated biofilm revealed a surprising spatial distribution of bacteria and genes, where potential polyphosphate accumulating organisms account for ∼ 47% of the inner layer microbiome. A spatial distribution of genes involved in dissimilatory nitrogen reduction was observed, indicating that different layers of the biofilm were metabolically active during the anoxic and aerobic zones of the phosphorus removal process. The physical biofilm separation into individual fractions unveiled functional layers of the biofilm, which will be important for future understanding of the phosphorus removal wastewater process.

Microbial ecological processes in MBBR biofilms for biological phosphorus removal from wastewater.

Phosphorus is both a major environmental pollutant and a limiting resource. Although enhanced biological phosphorus removal (EBPR) is used worldwide for phosphorus removal, the standard activated sludge-based EBPR process shows limitations with stability and efficiency. Recently, a new EBPR moving bed biofilm reactor (MBBR) process has been developed at HIAS (Hamar, Norway), enabling a phosphorus removal stability above 90% during a whole year cycle. To increase the knowledge of the HIAS (MBBR) process the aim of the current work was to characterize the MBBR microbiota and operational performance weekly for the operational year. Surprisingly, we found a major succession of the microbiota, with a five-fold increase in phosphorus accumulating organisms (PAOs), and major shifts in eukaryote composition, despite a stable phosphorus removal. Temperature was the only factor that significantly affected both phosphorus removal and the microbiota. There was a lower phosphor removal during the winter, coinciding with a higher microbiota alpha diversity, and a lower beta diversity. This differs from what is observed for activated sludge based EBPR. Taken together, the knowledge gained from the current microbiota study supports the efficiency and stability of MBBR-based systems, and that knowledge from activated sludge-based EBPR approaches cannot be translated to MBBR systems.