Transcriptional profiling elucidates biofilm functionality in the dynamic environment of an enhanced biological phosphorus removal reactor

ABSTRACT Recently, biofilms, complex and dynamic structures of microorganisms, have been applied to enhanced biological phosphorus removal (EBPR), a wastewater treatment configuration dependent on cyclic shifts between anaerobic and aerobic conditions. In this study, comparative metagenomics and metatranscriptomics were performed on biofilms collected from seven sites of a moving-bed-biofilm-reactor-based EBPR process. The aim was to examine the functional ecology of phosphorus-accumulating biofilms throughout a single EBPR cycle. Taxonomic profiling revealed high microbial diversity, stable throughout the EBPR cycle. The dominant phosphorus-accumulating organisms (PAOs) were identified as Candidatus accumulibacter, Candidatus phosphoribacter, and Candidatus lutibacillus. However, these did not show the highest transcriptional activities. Propionivibrio, a glycogen-accumulating organism, was the most transcriptionally active. Comparative analysis of biofilms from different EBPR stages showed a progressive change in metatranscriptome composition, correlating with nutrient removal. Analysis of differentially expressed genes in abundant PAOs revealed key genes associated with the uptake of phosphorus, degradation of glycogen, biosynthesis of polyhydroxyalkanoates, and acetate production. In conclusion, this study reveals that biofilms possess the capability to adapt to environmental fluctuations primarily through alterations in microbial gene expression activity and subsequent metabolic modulation, and dominant taxa may not necessarily exhibit the highest transcriptional activity in complex microbial communities.