Membrane bioreactors (MBRs) offer promising solution for wastewater treatment and reuse to address the problem of water scarcity. Nevertheless, this technology is still facing challenges associated with membrane biofouling. This phenomenon has been mainly investigated in lab-scale MBRs with little or no insight on biofouling in full-scale MBR plants. Furthermore, the temporal dynamics of biofouling microbial communities and their extracellular polymeric substances (EPS) are less studied. Herein, a multidisciplinary approach was adopted to address the above knowledge gaps in lab- and full-scale MBRs. In the full-scale MBR study, 16S rRNA gene pyrosequencing with multivariate statistical analysis revealed that the early and mature biofilm communities from five full-scale MBRs differed significantly from the source community (i.e. activated sludge), and random immigration of species from the source community was unlikely to shape the community structure of biofilms. Also, a core biofouling community was shared between the five MBR plants sampled despite differences in their operating conditions. In the lab-scale MBR studies, temporal dynamics of microbial communities and their EPS products were monitored on different hydrophobic and hydrophilic membranes during 30 days. At the early stages of filtration (1 d), the same early colonizers belonging to the class Betaproteobacteria were identified on all the membranes. However, their relative abundance decreased on day 20 and 30, and sequence reads belonging to the phylum Firmicutes and Chlorobi became dominant on all the membranes on day 20 and 30. In addition, the intrinsic membrane characteristic did not select any specific EPS fractions at the initial stages of filtration and the same EPS foulants developed with time on the hydrophobic and hydrophilic membranes. Our results indicated that the membrane surface characteristics did not select for specific biofouling communities or EPS foulants, and the same early colonizers were selected from the source community (i.e. activated sludge), and then went through significant changes to form a mature biofilm. Our findings from these studies could support future research aimed at developing enhanced biological-based strategies to control biofouling in MBRs.