First water safety plan approach applied to a Dental Clinic complex: identification of new risk factors associated with Legionella and P. aeruginosa contamination, using a novel sampling, maintenance and management program

Dental unit waterlines (DUWLs) represent a complex environment able to promote microbial contamination, due to functional, mechanical and practical risk factors. According to a water safety plan approach, the main goal is to preserve the health of dentists, dental staff and patients. The aim of this study is to develop a DUWLs water safety plan that is able to support correct and effective maintenance and disinfection procedures. Three different water systems serve 60 dental chairs: (i) water that comes directly from municipal water (Type A), (ii) water supplied by municipal water and water bottles (Type B) and (iii) water supplied only via water bottles (Type C). For each type, Legionella and Pseudomonas aeruginosa contamination was studied, by applying a new sampling scheme, based on separate sampling from water bottles, cup filler and handpieces. Type B DUWL is the only type of DUWL contaminated by L. pneumophila (ST 59) and L. anisa (mean contamination: 608.33 ± 253.33 cfu/L) detected in cup filler and handpieces, as well as the high presence of P. aeruginosa (44.42 ± 13.25 cfu/100 mL). Two subsequent shock treatments and resampling procedures were performed by increasing disinfectant dosage and contact time and removing some DUWL components linked to biofilm growth in DUWLs. A significant reduction of contamination was obtained for both microorganisms (Legionella spp.: −100%, p < 0.001 and P. aeruginosa: −99.86%, p = 0.006). The sampling strategy proposed allows us to identify the source of contamination and better focus on the maintenance and disinfection procedures. DUWLs represent an environment that requires a multidisciplinary approach, combining the knowledge of all DUWL components to correct procedures that are able to preserve the health of personnel and patients, as well as guaranteeing DUWLs’ safe functionality

Cooling towers influence in an urban environment: A predictive model to control and prevent Legionella risk and Legionellosis events

Cooling towers (CTs) are used to dissipate excess heat from water by evaporation, common in large facilities as hospital, companies, and hotels. The main risk attributed to CTs is represented by Legionella, a Gram-negative bacterium associated with a severe form of pneumonia known as Legionnaires' disease (LD). The infection route is by inhalation of aerosols reaching the lower respiratory tract. Despite several events associated with CTs, the knowledge in this field is still limited. The aim of this study was to develop a predictive model of bioaerosol dispersion using PM10 particles as a proxy, to generate risk maps of Legionella spread in the surrounding area in several weather and microbiological conditions. The Legionella contamination in the CT basin was 40938 ± 24523 cfu/L, with four peaks independent of the season, associated with an increase in air minimum temperature values (+1–2 °C) and a high relative humidity (66–100%) preceded by rainfall (0.2–30.6 mm/day). The model revealed that the most extensive bioaerosol spread is predicted in winter and summer, with an increase in Legionella risk at a distance of up to 1.5 km from the CT. This method represents a novel integrated approach for the prevention and management of LD risk in CTs

Development of a cooling tower predictive aerosol dispersion model to prevent Legionella infection

Cooling towers (CTs) are widely used devices in hospitals, industries, and other settings to dissipate heat from water via evaporation. The use of large amounts of water, stagnant conditions and poor maintenance programs promote the proliferation of bacteria such as Legionella . Since the 1980s, Legionella -containing aerosols produced by CTs have been linked to a number of outbreaks. To date, no studies have been conducted to assess the risk associated with Legionella dispersion through the CTs circuit, much less the preventive models that trace the map of its spread. The aim of the study is to develop a predictive model of bioaerosol dispersion linked to PM10, to develop a map of Legionella risk in a radius of 5-10 km from the hospital’s CT. This model represents a novel approach to the Legionella risk in CTs. Furthermore, the model could already be used in the design and installation phases, in addition to managing the CTs maintenance required to avoid outbreaks

Characterization of a Novel Species of <i>Legionella</i> Isolated from a Healthcare Facility: <i>Legionella resiliens</i> sp. nov

Two Legionella-like isolates, 8cVS16T and 9fVS26, were isolated from a water distribution system (WDS) in a healthcare facility. Cells were Gram- and Ziehl Neelsen-stain-negative, rod-shaped, motile, and exhibited a blue-white fluorescence under Wood’s lamp at 365 nm. The strains grew in a range of 32–37 °C on BCYE with L-cysteine (Cys+), GVPC, and MWY agar medium, with a positive reaction for oxidase, catalase, and gelatinase. The dominant fatty acids were summed features 3 (C16:1ω7c/C16:1ω6c) (27.7%), C16:0 iso (17.5%), and C16:0 (16.3%), and Q13 as the major ubiquinone. The mip and rpoB gene sequences showed a similarity of 96.7% and 92.4%, with L. anisa (ATCC 35292T). The whole genomes sequencing (WGS) performed displayed a GC content of 38.21 mol% for both. The digital DNA-DNA hybridization (dDDH) analysis demonstrated the separation of the two strains from the phylogenetically most related L. anisa (ATCC 35292T), with ≤43% DNA-DNA relatedness. The Average Nucleotide Identity (ANI) between the two strains and L. anisa (ATCC 35292T) was 90.74%, confirming that the two isolates represent a novel species of the genus Legionella. The name proposed for this species is Legionella resiliens sp. nov., with 8cVS16T (=DSM 114356T = CCUG 76627T) as the type strain