How Molecular Typing Can Support Legionella Environmental Surveillance in Hot Water Distribution Systems: A Hospital Experience.

In this study, we aimed to associate the molecular typing of Legionella isolates with a culture technique during routine Legionella hospital environmental surveillance in hot water distribution systems (HWDSs) to develop a risk map able to be used to prevent nosocomial infections and formulate appropriate preventive measures. Hot water samples were cultured according to ISO 11731:2017. The isolates were serotyped using an agglutination test and genotyped by sequence-based typing (SBT) for Legionella pneumophila or macrophage infectivity potentiator (mip) gene sequencing for non-pneumophila Legionella species. The isolates' relationship was phylogenetically analyzed. The Legionella distribution and level of contamination were studied in relation to temperature and disinfectant residues. The culture technique detected 62.21% of Legionella positive samples, characterized by L. pneumophila serogroup 1, Legionella non-pneumophila, or both simultaneously. The SBT assigned two sequence types (STs): ST1, the most prevalent in Italy, and ST104, which had never been isolated before. The mip gene sequencing detected L. anisa and L. rubrilucens. The phylogenetic analysis showed distinct clusters for each species. The distribution of Legionella isolates showed significant differences between buildings, with a negative correlation between the measured level of contamination, disinfectant, and temperature. The Legionella molecular approach introduced in HWDSs environmental surveillance permits (i) a risk map to be outlined that can help formulate appropriate disinfection strategies and (ii) rapid epidemiological investigations to quickly identify the source of Legionella infections.

Advances in Legionella Control by a New Formulation of Hydrogen Peroxide and Silver Salts in a Hospital Hot Water Network.

Legionella surveillance is an important issue in public health, linked to the severity of disease and the difficulty associated with eradicating this bacterium from the water environment. Different treatments are suggested to reduce Legionella risk, however long-term studies of their efficiency are lacking. This study focused on the activity of a new formulation of hydrogen peroxide and silver salts, WTP828, in the hospital hot water network (HWN) to contain Legionella contamination during two years of treatment. The effectiveness of WTP828 was tested measuring physical-chemical and microbiological parameters such as Legionella, Pseudomonas aeruginosa (P. aeruginosa), and a heterotopic plate count (HPC) at 36 °C. Legionella isolates were identified by serotyping and genotyping. WTP 828 induced a reduction in Legionella-positive sites (60% to 36%) and contamination levels (2.12 to 1.7 log10 CFU/L), with isolates belonging to L. pneumophila SG1 (ST1 and ST104), L. anisa and L. rubrilucens widely distributed in HWN. No relevant contamination was found for other parameters tested. The long-term effect of WTP828 on Legionella containment suggest the easy and safe application of this disinfectant, that combined with knowledge of building characteristics, an adequate environmental monitoring and risk assessment plan, become the key elements in preventing Legionella contamination and exposure.

Comparison between Two Types of Dental Unit Waterlines: How Evaluation of Microbiological Contamination Can Support Risk Containment.

Infection risk management in a dental unit waterline (DUWL) involves healthcare personnel and patients and is related to routine exposure to water and aerosols that may contain bacterial species. To improve water safety plans, maintenance, and sanitation procedures, analyses of heterotrophic plate counts (HPCs) at 36 °C, and two other microorganisms frequently associated with biofilms, Pseudomonas aeruginosa and Legionella spp., were performed in order to evaluate differences in microbiological contamination between two types of DUWLs: Type A, provided by a water tank, and Type B, directly connected to municipal water. The data showed that the water supply and water safety plan differentially influenced microbiological contamination: Type A DUWLs were more contaminated than Type B DUWLs for all microbiological parameters tested, with significant changes in the percentage of positive samples and contamination levels that were beyond the limits of standard guidelines. The results obtained show how the storage tank, the absence of anti-retraction valves, and the disinfection procedures performed are the main critical points of Type A DUWLs, which confirms that dental unit management (maintenance/sanitization) is often missed or not correctly applied by stakeholders, with an underestimation of the real risk of infection for patients and operators.