This study developed a magnetic nanoparticles (MNPs) harvesting and reactivation technique for rapid cyanobacteria Microcystis aeruginosa separation. The harvesting of raw MNPs achieved high efficiency of 99.6% with the MNPs dosage of 0.58g MNPs/g dry-biomass, but gradually decreased to 59.1% when directly reused 5 times. With extra ultrasonic chloroform:methanol solvent treatment, the MNPs can be effectively reactivated for M. aeruginosa harvesting with 60% efficiency after 5 times reactivation and the separation efficiency kept above 93% with 0.20g MNPs/g dry-biomass dosage. The cyanobacteria-MNPs complex can be effectively disrupted by ultrasonic chloroform:methanol solvent treatment and the zeta potential was recovered for MNPs electrostatic attraction. The MNPs adsorption followed the Langmuir isotherm, and the maximum adsorption capacity and Langmuir constant was 3.74g dry-biomass/g and 311.64L/g respectively. This MNPs reactivation technique can achieve low energy separation and reduce MNPs consumption by 67%, providing potential engineering implementation for cyanobacterial biomass harvesting.