Chlamydia-like obligate parasite of free-living amoebae.
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Biomedical subjects
Publications and source records attributed to T J Rowbotham.
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Following a recent phylogenetic analysis of all 39 species of the genus Legionella and related organisms, the name Legionella lytica has been proposed for bacteria previously considered either as Sarcobium lyticum or as 'Legionella-like amoebal pathogens' (LLAPs). To investigate the phylogenetic integrity of this newly proposed species, we determined the 16S rRNA gene sequences of 10 LLAPs isolated from various environmental sources. All 10 isolates clustered within a monophyletic group containing all other members of the genus Legionella. Eight of the 10 isolates formed a monophyletic subgroup within the genus which also included the two previously characterized L. lytica strains. Four of these 10 isolates shared a specific and very close relationship with the L. lytica type strain (> 99% sequence similarity). However, although clearly legionellae, the remaining two LLAP strains bore no specific evolutionary relationship to either L. lytica or any other Legionella species (< 96% sequence similarity). Both isolates lay on their own relatively deep-rooted branches within the radiation of the Legionella cluster. LLAPs do not, therefore, represent a unique species or even a single line of descent within the genus, and investigation of more isolates may reveal them to be as evolutionarily diverse as the other presently recognized Legionella species.
DNA coding for the 16S rRNA of an intracellular bacterium was directly amplified from lysed cells of a host amoebae using the polymerase chain reaction and primers specific for eubacteria. The amoebae had been used to recover an uncultured bacterium observed in the sputum of a patient with pneumonia. The amplified DNA was sequenced directly and compared with published 16S rRNA sequences. The analysis revealed that the intracellular bacterium is a member of the genus Legionella and that it is different from species, including L. pneumophila, for which 16S ribosomal RNA sequence data are available.
In the investigation of a large outbreak of non-pneumonic legionellosis at a leisure complex in Lochgoilhead, Scotland all direct cultures of environmental samples were initially negative for legionellae. Legionellae readily infect appropriate protozoa under suitable conditions, and following immunofluorescence to select specimens for special study, Legionella micdadei was isolated from whirlpool water via co-cultivation with Acanthamoeba polyphaga. L micdadei was also isolated, along with host amoebae, from the whirlpool filter. The use of amoebae in the isolation of legionellae from environmental (and other) sources can be of great value, especially if specimens shown by indirect immunofluorescence to contain legionellae fail to yield legionellae on routine culture.
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Amoebae are natural hosts for legionellae. The significance of this with regard to the multiplication of legionellae in the environment, the protection of legionellae from biocides, and the epidemiology of Legionnaires' Disease and Pontiac fever are discussed. A vesicle, or small amoeba, containing many legionellae is suggested as the main infective particle for man. We believe Pontiac fever can be explained as involving hypersensitivity to acanthamoebae, and comment on the lack of phages for legionellae. Legionella-like amoebal pathogen 1 (LLAP-1) was isolated, and probably belongs to a new species of Legionella. Further amoebal hosts for legionellae were found.
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The development of an amoebal enrichment method, for the recovery of viable Legionella pneumophila from clinical materials is described. The method has been used successfully in five of six cases of Legionnaires' disease in which L pneumophila was isolated. Studies on those L pneumophila isolates led to the discovery that virulent legionellae are attracted to, and attack suitable host amoebae, rather than infection following chance ingestion.
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Legionella pneumophila, the causative organism of Legionnaires' disease, is pathogenic for free living, ubiquitous, freshwater, and soil amoebae of the genera Acanthamoeba and Naegleria. Some species support the growth of strains from serogroups 1 to 6, others only strains from certain serogroups. Initial studies with seeded material indicate that amoebal enrichment could be utilised for the isolation of legionellae from clinical specimens and natural habitats. It is suggested that a vacuole, or amoeba, full of legionellae, rather than free legionellae, could be the infective particle for man.