The nonculturable form of Legionella pneumophila in multiple-nutrient starvation culture was studied. During extended starvation, the total direct counts (TDC) of L. pneumophila did not change significantly, but no colonies were detected on day 50. Quantitative PCR detection of L. pneumophila DNA demonstrated that nonculturable cells retained PCR-detectable DNA even after starvation for 300 days, and part of the nonculturable population possessed nonspecific esterase activity. However, resuscitation trials of nonculturable L. pneumophila on day 100 of starvation recovered no colony-forming units, and electrophoresis of nucleic acids extracted from nonculturable cells revealed rRNA subunit (23S, 16S and 5S) degradation. When legionellae have lost the ability to multiply, at least some DNA and enzyme functions may be retained for prolonged periods.