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Journal of Bacteriology, December 2002, p. 7025-7041, Vol. 184, No. 24
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.24.7025-7041.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Ultrastructural Analysis of Differentiation in Legionella pneumophila

Gary Faulkner1 and Rafael A. Garduño1,2*

Department of Microbiology and Immunology,1 Department of Medicine, Division of Infectious Diseases, Faculty of Medicine, Dalhousie University, Halifax, Nova Scotia B3H-4H7, Canada2

Received 14 May 2002/ Accepted 19 September 2002

Legionella pneumophila is an adaptive pathogen that replicates in the intracellular environment of fundamentally divergent hosts (freshwater protozoa and mammalian cells) and is capable of surviving long periods of starvation in water when between hosts. Physiological adaptation to these quite diverse environments seems to be accompanied by morphological changes (Garduño et al., p. 82-85, in Marre et al., ed., Legionella, 2001) and conceivably involves developmental differentiation. In following the fine-structural pathway of L. pneumophila through both in vitro and in vivo growth cycles, we have now discovered that this bacterium displays an unprecedented number of morphological forms, as revealed in ultrathin sections and freeze-fracture replicas for transmission electron microscopy. Many of the forms were identified by the obvious ultrastructural properties of their cell envelope, which included changes in the relative opaqueness of membrane leaflets, vesiculation, and/or profuse invagination of the inner membrane. These changes were best documented with image analysis software to obtain intensity tracings of the envelope in cross sections. Also prominent were changes in the distribution of intramembranous particles (clearly revealed in replicas of freeze-fractured specimens) and the formation of cytoplasmic inclusions. Our results confirm that L. pneumophila is a highly pleomorphic bacterium and clarify some early observations suggesting sporogenic differentiation in L. pneumophila. Since morphological changes occurred in a conserved sequence within the growth cycle, our results also provide strong evidence for the existence of a developmental cycle in L. pneumophila that is likely accompanied by profound physiological alterations and stage-specific patterns of gene expression.

* Corresponding author. Mailing address: Department of Microbiology & Immunology, Sir Charles Tupper Medical Building, 7th floor, Dalhousie University, 5859 University Avenue, Halifax, Nova Scotia, Canada B3H-4H7. Phone: (902) 494-6575. Fax: (902) 494-5125. E-mail: rafael.garduno{at}dal.ca.

Journal of Bacteriology, December 2002, p. 7025-7041, Vol. 184, No. 24
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.24.7025-7041.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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