Screening-Level Assays for Potentially Human-Infectious Environmental Legionella spp.

Helen Y. Buse; Abby Brehm; Jorge W. Santo Domingo; Nicholas J. Ashbolt

doi:10.1007/s12275-011-0233-z

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Screening-Level Assays for Potentially Human-Infectious Environmental Legionella spp.: Helen Y. Buse ¹, Abby Brehm ¹, Jorge W. Santo Domingo ², Nicholas J. Ashbolt ¹; Journal of Microbiology 2011;49(2):200-207.
DOI: https://doi.org/10.1007/s12275-011-0233-z
Published online: May 3, 2011

¹National Exposure Research Laboratory, Office of Research and Development, US Environmental Protection Agency, Cincinnati, OH 45268, USA, ²National Risk Management Research Laboratory, Office of Research and Development, US Environmental Protection Agency, Cincinnati, OH 45268, USA

Corresponding author: Helen Y. Buse , Tel: +1-513-569-7930,

Received: 29 June 2010 • Accepted: 8 December 2010

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Abstract

In spite of the fact that various Legionella species are isolated from nonclinical water settings, there is no standard method to determine whether environmental legionellae may be infectious to humans. Here we provide a screening-level approach based on an in vivo murine (A/J mouse) model and three in vitro proliferation assays using Acanthamoeba polyphaga, and THP-1 human and J774 murine macrophage cell lines to identify potentially human-infectious legionellae. As an initial demonstration the infectivity potential of three clinical (Legionella pneumophila, L. longbeacheae, and L. micdadei) and three environmental (L. dumoffii, L. maceachernii, and L. sainthelensi) legionellae were evaluated. A/J mice were intranasally infected and by 6 h post infection (p.i.), there were significant bacterial titers in the lungs. L. pneumophila, L. dumoffii, and L. micdadei densities were higher than L. longbeacheae, L. maceacherni, and L. sainthelensi at 24 h p.i. However, only L. pneumophila and L. micdadei persisted in the lungs after 48 h, indicating that the other isolates were rapidly cleared. Results from the in vitro assays showed that only L. pneumophila significantly multiplied within A. polyphaga, THP-1 and J774 cells after 72 h, but lysis of any of the in vitro hosts also flagged the strains for potential concern (e.g. L. dumoffii and L. micdadei). The results demonstrate the value of using multiple approaches to assess the potential level of pathogenicity of Legionella strains isolated from different environmental matrices.

Keywords: Legionella spp.;Acanthamoeba polyphaga;biofilms;virulence;macrophages

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Screening-Level Assays for Potentially Human-Infectious Environmental Legionella spp.

J. Microbiol. 2011;49(2):200-207. Published online May 3, 2011

DOI: https://doi.org/10.1007/s12275-011-0233-z

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