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A multicentric Italian investigation on legionnaires' disease is in course to clarify host factors as well as pathogen associated characteristics involved in the infection/disease. The main goal of the research plan is to account for some critical aspects concerning identification and prevention of legionellosis. To improve knowledge on factors associated with Legionella spp colonisation in hot waters, to detect cases and to characterize risk factors in subjects which develop pneumonia are specific objectives of the research programme. Preliminary results show that hot waters of houses and hotels are frequently contaminated (22.6% and 54.6%, respectively), mainly by L. pneumophila. Microbial concentrations were low in domestic waters (1.000 ufc/l), but higher in samples from the hotels (geom. mean 1.85 x 10(3) ufc/l). Warming system, age of the plant, type of building were risk factors significantly associated with Legionella spp positivity. The active surveillance on patients affected by pneumonia with search for Legionella urinary antigen allowed the identification of 34 cases, 3 of which of nosocomial origin, corresponding to 4.2% of the screened pneumonia. After informed consent, 26 subjects were recruited for a case-control-study to clarify risk factors for the disease.

The relationship between Legionella and Pseudomonas spp contamination and mineral content of domestic hot waters was investigated in a cross-sectional Italian survey. Pseudomonas spp contamination was associated with elevated Ca, but lower Fe and free chlorine content in the water. Waters with Cu concentrations50 microg/L prevented Legionella spp colonisation (OR = 0.14, 95% CI = 0.02-1.13), whereas the risk of legionellae occurrence increased in waters with Mn levels3 microg/L (OR = 2.37, 95% CI = 1.06-5.30). Furthermore, Mn was positively associated with the risk of colonisation by eterotrophic bacteria growing at either 22 and 36 degrees C. Legionella species and serogroups were differently distributed according to Cu and Mn concentrations, suggesting that Legionella strains may have a different sensibility/resistance to trace elements. A specific action of Cu as decontamination factor is suggested and the consideration of Mn as a risk indicator for bacterial colonisation and biofilm presence is proposed.

A cross-sectional multicenter survey of Italian hotels was conducted to investigate Legionella spp. contamination of hot water. Chemical parameters (hardness, free chlorine concentration, and trace element concentrations), water systems, and building characteristics were evaluated to study risk factors for colonization. The hot water systems of Italian hotels were strongly colonized by Legionella ; 75% of the buildings examined and 60% of the water samples were contaminated, mainly at levels of ≥10 3 CFU liter −1 , and Legionella pneumophila was the most frequently isolated species (87%). L. pneumophila serogroup 1 was isolated from 45.8% of the contaminated sites and from 32.5% of the hotels examined. When a multivariate logistic model was used, only hotel age was associated with contamination, but the risk factors differed depending on the contaminating species and serogroup. Soft water with higher chlorine levels and higher temperatures were associated with L. pneumophila serogroup 1 colonization, whereas the opposite was observed for serogroups 2 to 14. In conclusion, Italian hotels, particularly those located in old buildings, represent a major source of risk for Legionnaires' disease due to the high frequency of Legionella contamination, high germ concentration, and major L. pneumophila serogroup 1 colonization. The possible role of chlorine in favoring the survival of Legionella species is discussed.

We investigated Legionella and Pseudomonas contamination of hot water in a cross-sectional multicentric survey in Italy. Chemical parameters (hardness, free chlorine, and trace elements) were determined. Legionella spp. were detected in 33 (22.6%) and Pseudomonas spp. in 56 (38.4%) of 146 samples. Some factors associated with Legionella contamination were heater type, tank distance and capacity, water plant age, and mineral content. Pseudomonas presence was influenced by water source, hardness, free chlorine, and temperature. Legionella contamination was associated with a centralized heater, distance from the heater point >10 m, and a water plant >10 years old. Furthermore, zinc levels of 50 microg/L appeared to be protective against Legionella colonization [corrected]. Legionella species and serogroups were differently distributed according to heater type, water temperature, and free chlorine, suggesting that Legionella strains may have a different sensibility and resistance to environmental factors and different ecologic niches.

Several pathologies of respiratory tract are referred to be related to bacteria belonging to Legionella genus1. This pathology mainly affects immunocompromized people as nosocomial or community patients. Surveillance programs are required for monitoring bacteria diffusion and optimization of periodic decontamination action. Detection of Legionella pneumophila in water is based on well-standardized cultural and morphological analysis indicated in national guidelines2. Species assignment is typically performed by rapid agglutination tests3, which can be realized in few minutes. They usually provide univocal results in discrimination and serogrouping of L. pneumophila versus L. species including L. longbeachae, L. bozemanii, L. dumofii, L. jordanis, L. micdadei and L. anisa. This test is based on recognition of lipopolysaccharide present on the outer membrane4,5. We report some evidences where phenotypic procedures failed. During surveillance programs6,7 some unclearly identified bacteria from different regions were isolated. They showed morphological and cultivation features of legionellaceae but species classification, performed by agglutination test, gave negative or equivocal results (Figure 1). In particular they provide an absent or ambiguous agglutination of latex particles as can be observed in the representative Figure 1 (panels 1 and 3) where, positive results were obtained from a clear formation of precipitates (Figure 1, panel 2), as suggested by manufacturing practices and described by Dennis8 and related references. In principle this kind of equivocal results have to be considered negative. Bacteria that showed these results were definitely identified by DNA sequencing.