Legionella is an environmental pathogen that is responsible for respiratory disease and is a common causative agent of water-related outbreaks. Due to their ability to survive in a broad range of environments, transmission of legionellosis is possible from a variety of sources. Unfortunately, a disproportionate amount of research that is devoted to studying the occurrence of Legionella in environmental reservoirs is aimed toward cooling towers and premise plumbing. As confirmed transmission of Legionella has been linked to many other sources, an over-emphasis on the most common sources may be detrimental to increasing understanding of the spread of legionellosis. This review aims to address this issue by cataloguing studies which have examined the occurrence of Legionella in less commonly investigated environments. By summarizing and discussing reports of Legionella in fresh water, ground water, saltwater, and distribution system drinking water, future environmental and public health researchers will have a resource to aid in investigating these pathogens in relevant sources.

As a waterborne pathogen of increasing concern, techniques for cost-effective and rapid characterization of Legionella are vital. This study examines the development of a Matrix Assisted Laser Desorption/Ionization-Time of Flight Mass Spectrometry (MALDI-TOF-MS) analysis methodology for this microbe. First, optimal sample preparation methods for the analysis of environmental Legionella isolates via MALDI-TOF-MS were determined. These methods were then implemented to perform strain-level characterization of environmental Legionella isolates from central Arizona. Results demonstrate that a MALDI-TOF-MS method involving BCYE agar-based culturing and protein extraction-based sample preparation yield high-quality mass spectra. Twenty-eight environmental Legionella isolates originating from two separate drinking water distribution systems were analyzed. Multiple species were detected, and strain-level characterization was achieved, with 12 unique strains distinguished. In addition, isolates of L. pneumophila, the most common species observed in the study, were correctly assigned to specific sampling sites. These results demonstrate the potential for this technique to be applied for sub-species characterization of Legionella with significant benefits over established methodologies.