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The time between two consecutive samples was dependent on the sampling frequency, and could range from 2 to 7?days depending on the site. Real-time PCR and culture ratios (r) were calculated for http://www.selleckchem.com/products/3-methyladenine.html two consecutive samples as follows: PCR ratio?=?(log GU?l?1 at day n)/(log GU?l?1 at day n-1); culture ratio?=?(log CFU?l?1 at day n)/(log CFU?l?1 at day n-1). For each ratio, the following values were analysed: (i) r?=?1 meant that no changing occurred between two consecutive values; (ii) r??1 meant that an increase occurred between two consecutive values. To evaluate if there were significant differences between culture and real-time PCR ratios for each system, a statistical Wilcoxon test was performed (P? http://www.selleck.cn/products/Cisplatin.html by real-time PCR as a management tool to monitor concentrations of Legionella spp. and L. pneumophila in cooling systems and to support for the cultural standard method AFNOR T90-431, quantifications were achieved for nine cooling systems with different water qualities and for a total of 185 samples from August to December 2006 (Table?1). As expected from the high selectivity of the GVPC medium for L. pneumophila by the culture method (Lee et?al. 1993; Ta et?al. 1995; Bartie et?al. 2003, Riffard et?al., 2003), almost all the numerations of Legionella spp. have given the same concentrations as L. pneumophila (representing 83�C100% of the isolated strains), irrespective of the cooling system analysed. They ranged from the detection limit (except A3 and D1: 5��7?��?103 and 6��6?��?103?CFU?l?1 respectively) to 9��5?��?103?CFU?l?1 (deionized water), to 3��5?��?104?CFU l?1 (river water submitted to a biocide http://www.selleckchem.com/products/PD-0325901.html treatment) and to 1��2?��?106?CFU l?1 (river water). By this observation, we considered that the culture method was of interest for numeration of L. pneumophila only. Hence, only L. pneumophila counts were finally compared with the concentrations measured by the real-time PCR. Among the 165 samples analysed by culture, 23 of them have not been quantified due to the interfering flora, even with the heat and acid treatments. This problem was principally found in the samples of site A provided by raw river water (19/23), but among them some samples have finally been quantified (7/19), after a 10-fold dilution step. As shown in Fig.?1, Legionella spp. and L. pneumophila were quantified by real-time PCR in higher concentrations than by the culture method. In river water samples (sites A, B and D), Legionella spp. were quantified from the quantification limit (3��4?��?102?GU?l?1) to 1��6?��?107?GU?l?1 and L. pneumophila from the quantification limit (3��4?��?102?GU?l?1) to 2��9?��?106?GU?l?1.