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Applied and Environmental Microbiology, June 2002, p. 3076-3084, Vol. 68, No. 6
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.6.3076-3084.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Crescent Bodies of Parachlamydia acanthamoeba and Its Life Cycle within Acanthamoeba polyphaga: an Electron Micrograph Study

Gilbert Greub and Didier Raoult*

Unité des Rickettsies, Faculté de Médecine, Université de la Méditerranée, Marseille, France

Received 27 September 2001/ Accepted 17 December 2001

Parachlamydiaceae are endosymbionts of free-living amoeba first identified in 1997. Two developmental stages, elementary and reticulate bodies, were observed; however, their localization and proportions according to culture condition and duration remain unknown. The life cycle of Parachlamydia acanthamoeba within Acanthamoeba polyphaga was studied by transmission electron microscopy of 8-, 36-, and 144-h coculture. Morphometry and quantification were performed using SAMBA software. The elementary body, the predominant stage within the amoebae, was located mainly within their vacuoles. The multiplication of Parachlamydia bacteria by binary fission of reticulate bodies was independently associated with culture in PYG broth (odds ratio [OR] = 4.4; 95% confidence interval [CI], 1.55 to 12.46) and with the presence of reticulate bodies within the amoebae (OR = 2.10; 95% CI, 1.53 to 2.89). A third developmental stage was observed, the crescent body. Its presence outside and inside the amoebae was associated mainly with prolonged incubation time (OR = 3.98; 95% CI, 1.49 to 10.68, and OR = 5.98; 95% CI, 1.75 to 20.4, respectively). Elementary and crescent bodies were released into the extracellular medium within vesicles or after amoebal lysis. For both, phagocytosis was their mode of entry. This electron micrograph study revealed another infective developmental stage, the crescent body, and provided quantitative analysis of the life cycle of P. acanthamoeba within A. polyphaga.

* Corresponding author. Mailing address: Unité des Rickettsies, CNRS UPRESA 6020, Faculté de médecine, Université de la Méditerranée, 27 Blvd. Jean Moulin, 13385 Marseille Cedex 05, France. Phone: 00 33 491 32 43 75. Fax: 00 33 491 83 03 90. E-mail: didier.raoult{at}medecine.univ-mrs.fr.

Applied and Environmental Microbiology, June 2002, p. 3076-3084, Vol. 68, No. 6
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.6.3076-3084.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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