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• The effects of anionic, cationic and neutral organic matters on the aggregation of microplastics are investigated. • All of the organic matters show positive effects on the growth of biofilm-coated microplastic aggregates. • The two-class population balance model is used to track the small and large microplastic subgroups under turbulence. Biofilm-coated microplastics are omnipresent in aquatic environments, carrying different organic matter (OM) that in turn influences the flocculation and settling of microplastic aggregates. In this study, the effects of chitosan, guar gum, humic acid, and xanthan gum on the flocculation of anthropogenic microplastics are examined under controlled shear through the mixing chamber experiments. The results show that all of the selected OMs have positive effects on biofilm culturing and thus enhance the growth of microplastic flocs, with more evident promoting effects for cationic and neutral OMs (i.e., chitosan and guar gum) than anionic OMs (i.e., humic acid and xanthan). No critical shear rate is observed in the size vs. shear relationship based on our measurements. In addition, the quadrature-based two-class population balance model is employed to track the development of bimodal floc size distributions (FSDs) composed of small and large microplastic flocs. The model predictions show reasonable agreement with the observed FSDs. The largest error of settling flux from the two-class model is 7.8% in contrast with the reference value measured by the camera-based FSDs with 30 bins. This study highlights the role of different OMs on microplastic flocculation and indicates that a two-class model may be sufficient to describe microplastic transport processes in estuaries. [Display omitted] [ABSTRACT FROM AUTHOR]