SUMMARYEarlier studies on the ecology of leptospirosis in temperate regions focused mainly on free-ranging rats in rural areas. Here we report on the occurrence of Leptospira spp. in Rattus norvegicus living in sewers in a suburban area in Copenhagen, Denmark. In 2006-2007, about 30 rats were captured in sewers at each of six different locations. Rat kidneys were screened by PCR for pathogenic Leptospira spp. In one location no infected rats were found, whereas the prevalence in the remaining five locations ranged between 48% and 89%. Micro-agglutination tests showed that serogroup Pomona, Sejroe, and Icterohaemorrhagiae were the most common. Infection was related to age with the highest prevalence observed for adult rats but there was no difference in infection rate between sexes, suggesting primarily environmental transmission. Since most reported rat problems in urban areas are related to sewer rats, the surprisingly high level of infection calls for an increased public health concern.