Abstract
Legionnaires’ disease incidence is on the rise, with the majority of cases attributed to the intracellular pathogen, Legionella pneumophila. Nominally a parasite of protozoa, L. pneumophila can also infect alveolar macrophages when bacteria-laden aerosols enter the lungs of immunocompromised individuals. L. pneumophila pathogenesis has been well characterized; however, little is known about the >25 different Legionella spp. that can cause disease in humans. Here, we report for the first time a study demonstrating the intracellular infection of an L. bozemanae clinical isolate using approaches previously established for L. pneumophila investigations. Specifically, we report on the modification and use of a green fluorescent protein (GFP)-expressing plasmid as a tool to monitor the L. bozemanae presence in the Acanthamoeba castellanii protozoan infection model. As comparative controls, L. pneumophila strains were also transformed with the GFP-expressing plasmid. In vitro and in vivo growth kinetics of the Legionella parental and GFP-expressing strains were conducted followed by confocal microscopy. Results suggest that the metabolic burden imposed by GFP expression did not impact cell viability, as growth kinetics were similar between the GFP-expressing Legionella spp. and their parental strains. This study demonstrates that the use of a GFP-expressing plasmid can serve as a viable approach for investigating Legionella non-pneumophila spp. in real time.
Funder
Natural Sciences and Engineering Research Council of Canada
Subject
Virology,Microbiology (medical),Microbiology
Reference49 articles.
1. Surveillance for Waterborne Disease Outbreaks Associated with Drinking Water — United States, 2013–2014
2. Australia’s Notifiable Disease Status, 2014: Annual Report of the National Notifiable Diseases Surveillance;System Commun. Dis. Intell.,2016
3. European Centre for Disease Prevention and Control (ECDC) Surveillance Atlas—Legionnaires Disease
https://www.ecdc.europa.eu/en/legionnaires-disease/surveillance/atlas
4. Public Health Agency of Canada Legionellosis Reported Cases from 1986 to 2018 in Canada
https://diseases.canada.ca/notifiable/charts?c=yl
5. Annual Epidemiological Report for 2017 Legionnaires’ Disease,2019
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