Abstract
A Gram-negative, rod-shaped bacterium, designated H63(T), was isolated from aortic valve tissue of a patient with native valve endocarditis. 16S rRNA gene sequencing revealed that H63(T) belongs to the genus Legionella, with its closest neighbours being the type strains of Legionella brunensis (98.8% similarity), L. londiniensis (97.0%), L. jordanis (96.8%), L. erythra (96.2%), L. dresdenensis (96.0%) and L. rubrilucens, L. feeleii, L. pneumophila and L. birminghamensis (95.7%). DNA-DNA hybridization studies yielded values of <70% relatedness between strain H63(T) and its nearest neighbours in terms of 16S rRNA gene sequence similarity, indicating that the strain represents a novel species. Phylogenetic analysis of the 16S rRNA, macrophage infectivity potentiator (mip) and RNase P (rnpB) genes confirmed that H63(T) represents a distinct species, with L. brunensis being its closest sister taxon. Fatty acid composition and biochemical traits, such as the inability to ferment glucose and reduce nitrate, supported the affiliation of H63(T) to the genus Legionella. H63(T) was distinguishable from its neighbours based on it being positive for hippurate hydrolysis. H63(T) was further differentiated by its inability to grow on BCYE agar at 17 °C, its poor growth on low-iron medium and the absence of sliding motility. Also, H63(T) did not react with antisera generated from type strains of Legionella species. H63(T) replicated within macrophages. It also grew in mouse lungs, inducing histopathological evidence of pneumonia and dissemination to the spleen. Together, these results confirm that H63(T) represents a novel, pathogenic Legionella species, for which the name Legionella cardiaca sp. nov. is proposed. The type strain is H63(T) ( = ATCC BAA-2315(T) = DSM 25049(T) = JCM 17854(T)).