A microbiological study for the isolation and characterization of Leptospira interrogans was performed in an attempt to define the characteristics of Leptospirosis in Korea. The results are summerised as follows. Thirty-five cultures were isolated from 11 patients with leptospirosis and 24 wild rodents captured in Paju area. The isolation rate of Leptospira from wild rodents reached 23.1%. All 35 cultures were identified as Leptospira interrogans by their characteristic morphology and motility in dark field microscopy, pathogenecity in Guinea pig and sensitivity to 8-Azaguanine. Cultures formed diffuse subsurface colonies with hazy margin and were catalase, peroxidase and oxdase positive. The fifty percent lethal dose of isolate HM 4 in Guinea pig was $8.9{ imes}10^4$ org. Mouse passage method was sucecssfully applied in maintenance of fresh isolates without any loss of their original virulency.