Phycosphere microbiome succession tracks algal growth dynamics during phosphorus-limited wastewater treatment

Algal-based wastewater treatment (phycoremediation) relies on microbial interactions within the algal phycosphere that are associated with algal growth and nutrient removal. However, the temporal dynamics of these communities under operational conditions remain poorly resolved. Using 16S and 18S rRNA gene metabarcoding, this study characterized bacterial and eukaryotic communities across alga-attached and free-living size fractions and over time in a raceway-based, pilot-scale phycoremediation system using a filamentous algal co-culture under semicontinuous municipal wastewater flow. Bacterial community composition in the phycosphere overlapped substantially with that observed in a previous laboratory-scale study using the same algal co-culture, with many highly abundant ASVs shared across studies, supporting consistency of key community members across scales. Phycosphere community dynamics were temporally aligned with algal growth, with bacterial alpha diversity in the alga-attached fraction highest during periods of active algal growth and declining with the onset of algal phosphorus limitation. During this high-diversity phase, several orders within Alphaproteobacteria were enriched, followed by declines as the algal culture progressed toward reduced growth. Eukaryotic communities also showed clear successional trends, with Perkinsids (Alveolata) increasing during peak algal biomass before giving way to diverse protists and rotifers. These findings demonstrate coordinated temporal patterns between algal growth dynamics and phycosphere microbial succession in pilot-scale wastewater raceways, providing operational insight into microbial community structure under phosphorus-limited phycoremediation. • Pilot-scale raceway cultivation under semicontinuous municipal wastewater flow • Phycosphere bacterial alpha diversity highest during active algal growth • Phosphorus limitation in algae precedes and drives phycosphere diversity decline. • Phycosphere composition overlaps with previous lab-scale study. • Protist succession shifts from perkinsids to diverse protists and rotifers.