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The incidence of Legionnaires’ disease (LD) in European countries and the USA has been constantly increasing since 1998. Infection of humans occurs through aerosol inhalation. To bridge the existing gap between the concentration of  Legionella  in a water network and the deposition of bacteria within the thoracic region (assessment of the number of viable  Legionella ), we validated a model mimicking realistic exposure through the use of (i) recent technology for aerosol generation and (ii) a 3D replicate of the human upper respiratory tract. The model’s sensitivity was determined by monitoring the deposition of (i) aerosolized water and Tc 99m  radio-aerosol as controls, and (ii) bioaerosols generated from both  Escherichia coli  and  Legionella pneumophila  sg 1 suspensions. The numbers of viable  Legionella  prior to and after nebulization were provided by culture, flow cytometry and qPCR. This study was designed to obtain more realistic data on aerosol inhalation ( vs . animal experimentation) and deposition at the thoracic region in the context of LD. Upon nebulization, 40% and 48% of the initial  Legionella  inoculum was made of cultivable and non-cultivable cells, respectively; 0.7% of both populations reached the filter holder mimicking the thoracic region in this setup. These results are in agreement with experimental data based on quantitative microbial risk assessment methods and bring new methods that may be useful for preventing LD.

Experimental human-like model to assess the part of viable Legionella reaching the thoracic region after nebulization