The Legionella colonization frequency at 385 Greek hotel hot and cold water distribution systems was 20.8%. Legionella contamination was associated with the presence of an oil heater (odds ratio [OR]=2.04, 95% confidence interval [CI]=1.12 to 3.70), with the sample temperature (OR=0.26, 95% CI=0.1 to 0.5), with seasonal operation (OR=3.23, 95% CI=1.52 to 6.87), and with the presence of an independent disinfection system (OR=0.30, 95% CI=0.15 to 0.62). The same water temperatures, free-chlorine levels, and pHs differently affect the survival of various Legionella spp.