Machine Learning Approach Reveals the Assembly of Activated Sludge Microbiome with Different Carbon Sources during Microcosm Startup.

Activated sludge (AS) microcosm experiments usually begin with inoculating a bioreactor with an AS mixed culture. During the bioreactor startup, AS communities undergo, to some extent, a distortion in their characteristics (e.g., loss of diversity). This work aimed to provide a predictive understanding of the dynamic changes in the community structure and diversity occurring during aerobic AS microcosm startups. AS microcosms were developed using three frequently used carbon sources: acetate (A), glucose (G), and starch (S), respectively. A mathematical modeling approach quantitatively determined that 1.7-2.4 times the solid retention time (SRT) was minimally required for the microcosm startups, during which substantial divergences in the community biomass and diversity (33-45% reduction in species richness and diversity) were observed. A machine learning modeling application using AS microbiome data could successfully (>95% accuracy) predict the assembly pattern of aerobic AS microcosm communities responsive to each carbon source. A feature importance analysis pinpointed specific taxa that were highly indicative of a microcosm feed source (A, G, or S) and significantly contributed for the ML-based predictive classification. The results of this study have important implications on the interpretation and validity of microcosm experiments using AS.