Prevalence and Characterization of Legionella pneumophila and Related Species from Water-Based Recreational Sites

Abstract Background and Objectives   Legionella is a genus of gram-negative bacteria common in soil and aquatic systems and is associated with protists. They have emerged as a pathogenic group due to the increased use and poor maintenance of artificial water environments. This study aims at phenotypic and molecular identification of Legionella in water and swab samples collected from water-based recreational sites. The biofilm-forming ability of Legionella on exposure to various temperatures and iron concentrations was also studied. Methods  A total of 60 samples including 30 swab samples and 30 water samples (decorative fountain ponds, swimming pools, garden sprinklers, drip irrigation system) were collected from in and around Mangalore, Karnataka, India. From each source, swab and water samples were collected as per the Indian standard IS: 1622. The collected samples were processed within 4 hours of collection. The samples were subjected to microbiological and chemical estimation followed by filtration through a 0.2 µm membrane filter. Isolation of Legionella from collected samples was performed as per US Centers for Disease Control and Prevention guidelines 2005. The positive isolates were then checked for biofilm-forming ability at various temperatures (25°C, 35°C, and 45°C) and iron concentration (3 mg/L, 30 mg/L, and 300 mg/L) using crystal violet assay. Results  Out of 30, water and swab samples tested, one water sample from a garden sprinkler, swimming pool, and one from both water and swab samples from a decorative fountain showed the presence of Legionella . A biofilm study of Legionella at various temperatures and iron concentrations categorized the bacteria as a moderate biofilm former. Conclusion  This study revealed that most of the water and swab samples were found to be negative for Legionella that is quite encouraging and the contamination of water systems in recreational facilities can be reduced by decontamination techniques and proper hygienic practices.