For the need of bio-degradable flocculant in stage of wastewater treatment, some cultural conditions of bioflocculant production were optimized with Aeromonas hydrophila KH-54. About 260 strains of type culture and bacteria isolated from marsh, pond, activated sludge, etc were examined for their ability to flocculate kaolin particles and swine wastewater. Among them, KH-54 showed the highest flocculating activity and was identified as Aeromonas hydrophila according to the cultural, morphological and physiological properties. The maximum production of the flocculant secreted by Aeromonas hydrophila KH-54 was observed in culture medium containing 2.0% mannitol, 0.05% ammonium chloride, 0.02% potassium phosphate dibasic, 0.01% MgSO₄· 7H₂O and 0.05% yeast extract at initial pH 7.0 when cultured on rotary shaker controlled at 25℃ and 150 rpm. Under the optimized condition, the flocculating ability reached to 770 units/㎖ of kaolin flocculating activity and 81% of NTU removal efficiency against swine wastewater after 4 days cultivation. The bioflocculant was also effective on various organic wastewaters other than swine wastewater, showing NTU removal rate ranging from 92% to 34%.