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Magnetic micro-particles (MMPs) and magnetic field (MF) were utilized as a coupling conditioning unit before dewatering activated sludge (AS) under a horizontal electric field. The removal and migration of free and bound water in AS during coupling magnetic conditioning–horizontal electro-dewatering (CM–HED) were determined. The organic matter migration between the solid and liquid phases of AS biosolids was also analyzed. Results show that MMPs dosage and MF intensity were determined as 0.15 g/g dry solids and 0.065 T for the best dewaterability, respectively. The optimum dewatering conditions for CM–HED with the final water content of 89.98% were 40 V and 120 min as determined using the response surface methodology. MMPs conditioning could induce a slight coagulation among AS flocs, increase the particle size from 85.9 μm to 92.3 μm and decrease mass fractal dimension from 2.18 to 2.07. The MMPs-conditioned AS also showed a network-like structure, banded cells with shrunk surfaces. CM–HED process effectively reduced the free water content (FWC) and bound water content (BWC) but increased the portion of BWC in AS. The corresponding removal ratios of bound water and free water were 52.89% and 95.86% at the anode side and 46.28% and 92.75% at the cathode side, respectively. The coupling magnetic conditioning led to the largest BWC reduction of 23.14% in CM–HED process, and most of this reduction approaching 92.83% occurred during magnetic micro-particle conditioning stage. Gravity field caused the largest sludge reduction of 87.45%. During HED stage, the removal ratio of free water in AS was 63.73% at the anode side and 36.54% at the cathode side, while it was 21.9% and 10.96% for bound water, respectively. Along with water removed by CM–HED process, the organic matter in supernatant/filtrate increased, and a sharp rise occurred during HED stage. Meanwhile, the extracellular polymeric substance (EPS) contents initially decreased at MMPs–MF conditioning stage and then increased at the HED stage. Lower EPS contents in AS indicated better dewaterability. The largest reduction in the EPS content of AS at MMPs–MF conditioning stage corresponded to the largest decline in BWC. Furthermore, the protein-like substances in the supernatant/filtrate increased from 30% to 50%, whereas the fulvic acid-like and humic acid-like substances decreased from 54% to 27% at the HED stage. MMPs significantly reduced the polysaccharide and protein contents in the slime–EPS by 90.26% and 99.25%, respectively. Approximately 92.35% of humic acid-like materials in tightly bound EPS were reduced by MMPs. Thus, coupling magnetic conditioning should be selected as pretreatment prior to HED. [ABSTRACT FROM AUTHOR]