In this study, a microcolony technique was combined with direct fluorescent antibody staining for the specific detection and enumeration of Legionella pneumophila in freshwater samples with growth activity. This method allowed the detection of active L. pneumophila (within 48 h) in 91 bath water samples collected from 30 bathing facilities, with similar sensitivity of a conventional plate-counting method. These results suggest that the microcolony method combined with fluorescent antibody staining could be useful as a monitoring technique for the prevention of Legionnaires' disease through the early detection of L. pneumophila in freshwater.