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Protochlamydia naegleriophila as etiologic agent of pneumonia.

Casson N, Michel R, Müller KD, Aubert JD, Greub G - Emerging Infect. Dis. (2008)

Bottom Line: Phenotypic, genetic, and phylogenetic analyses supported its affiliation as Protochlamydia naegleriophila sp. nov.We then developed a specific diagnostic PCR for Protochlamydia spp.When applied to bronchoalveolar lavages, results of this PCR were positive for 1 patient with pneumonia.

View Article: PubMed Central - PubMed

Affiliation: Center for Research on Intracellular Bacteria, Institute of Microbiology, University Hospital Center and University of Lausanne, Lausanne, Switzerland.

ABSTRACT
Using ameba coculture, we grew a Naegleria endosymbiont. Phenotypic, genetic, and phylogenetic analyses supported its affiliation as Protochlamydia naegleriophila sp. nov. We then developed a specific diagnostic PCR for Protochlamydia spp. When applied to bronchoalveolar lavages, results of this PCR were positive for 1 patient with pneumonia. Further studies are needed to assess the role of Protochlamydia spp. in pneumonia.

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Transmission electron microscopy of Protochlamydia naegleriophila. A) Naegleria lovaniensis trophozoite after transfer of endocytobionts; strain KNic (p) from the original host strain showing 15 coccoid bacteria distributed randomly within the cytoplasm of the host ameba. N, nucleus; en, endosome (karyosome) within the nucleus; v, empty food vacuoles. Magnification ×10,500; bar = 1 μm. B) N. lovaniensis trophozoite jammed with numerous endoparasitic stages of Pr. naegleriophila. Magnification ×16,800; bar = 1 μm. C) Enlarged detail of N. lovaniensis trophozoite with intracytoplasmic stages of Pr. naegleriophila. Some stages show binary fission indicated by the fission furrow (arrows). The endoparasites have a wrinkled gram-negative outer membrane rendering a spiny appearance to the endoparasites. Signs of damage are obvious within the cytoplasm of the host ameba. mi, mitochondria. Magnification ×43,500; bar = 0.5 μm. D) Pr. naegleriophila within vacuoles of Acanthamoebae castellanii ameba 2 days postinfection. Elementary bodies (eb) and reticulate bodies (rb) are visible. Elementary bodies harbor a smooth membrane compared with the reticulate bodies, which have a spiny shape. Magnification ×10,000; bar = 2 μm. E) Enlarged detail of A. castellanii trophozoite with intracytoplasmic stages of Pr. naegleriophila 3 days postinfection. Binary fission is observed (di). Magnification ×20,000; bar = 1 μm. F) Crescent body (arrow) within A. castellanii observed 3 days postinfection. Magnification ×20,000; bar = 1 μm.
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Figure 1: Transmission electron microscopy of Protochlamydia naegleriophila. A) Naegleria lovaniensis trophozoite after transfer of endocytobionts; strain KNic (p) from the original host strain showing 15 coccoid bacteria distributed randomly within the cytoplasm of the host ameba. N, nucleus; en, endosome (karyosome) within the nucleus; v, empty food vacuoles. Magnification ×10,500; bar = 1 μm. B) N. lovaniensis trophozoite jammed with numerous endoparasitic stages of Pr. naegleriophila. Magnification ×16,800; bar = 1 μm. C) Enlarged detail of N. lovaniensis trophozoite with intracytoplasmic stages of Pr. naegleriophila. Some stages show binary fission indicated by the fission furrow (arrows). The endoparasites have a wrinkled gram-negative outer membrane rendering a spiny appearance to the endoparasites. Signs of damage are obvious within the cytoplasm of the host ameba. mi, mitochondria. Magnification ×43,500; bar = 0.5 μm. D) Pr. naegleriophila within vacuoles of Acanthamoebae castellanii ameba 2 days postinfection. Elementary bodies (eb) and reticulate bodies (rb) are visible. Elementary bodies harbor a smooth membrane compared with the reticulate bodies, which have a spiny shape. Magnification ×10,000; bar = 2 μm. E) Enlarged detail of A. castellanii trophozoite with intracytoplasmic stages of Pr. naegleriophila 3 days postinfection. Binary fission is observed (di). Magnification ×20,000; bar = 1 μm. F) Crescent body (arrow) within A. castellanii observed 3 days postinfection. Magnification ×20,000; bar = 1 μm.

Mentions: A. castellanii/KNic and N. lovaniensis/KNic cocultures were processed for electron microscopy as described. Ameba filled with bacteria exhibiting 3 developmental stages already described in other Parachlamydiaceae (8) were observed (Figure 1).


Protochlamydia naegleriophila as etiologic agent of pneumonia.

Casson N, Michel R, Müller KD, Aubert JD, Greub G - Emerging Infect. Dis. (2008)

Transmission electron microscopy of Protochlamydia naegleriophila. A) Naegleria lovaniensis trophozoite after transfer of endocytobionts; strain KNic (p) from the original host strain showing 15 coccoid bacteria distributed randomly within the cytoplasm of the host ameba. N, nucleus; en, endosome (karyosome) within the nucleus; v, empty food vacuoles. Magnification ×10,500; bar = 1 μm. B) N. lovaniensis trophozoite jammed with numerous endoparasitic stages of Pr. naegleriophila. Magnification ×16,800; bar = 1 μm. C) Enlarged detail of N. lovaniensis trophozoite with intracytoplasmic stages of Pr. naegleriophila. Some stages show binary fission indicated by the fission furrow (arrows). The endoparasites have a wrinkled gram-negative outer membrane rendering a spiny appearance to the endoparasites. Signs of damage are obvious within the cytoplasm of the host ameba. mi, mitochondria. Magnification ×43,500; bar = 0.5 μm. D) Pr. naegleriophila within vacuoles of Acanthamoebae castellanii ameba 2 days postinfection. Elementary bodies (eb) and reticulate bodies (rb) are visible. Elementary bodies harbor a smooth membrane compared with the reticulate bodies, which have a spiny shape. Magnification ×10,000; bar = 2 μm. E) Enlarged detail of A. castellanii trophozoite with intracytoplasmic stages of Pr. naegleriophila 3 days postinfection. Binary fission is observed (di). Magnification ×20,000; bar = 1 μm. F) Crescent body (arrow) within A. castellanii observed 3 days postinfection. Magnification ×20,000; bar = 1 μm.
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Figure 1: Transmission electron microscopy of Protochlamydia naegleriophila. A) Naegleria lovaniensis trophozoite after transfer of endocytobionts; strain KNic (p) from the original host strain showing 15 coccoid bacteria distributed randomly within the cytoplasm of the host ameba. N, nucleus; en, endosome (karyosome) within the nucleus; v, empty food vacuoles. Magnification ×10,500; bar = 1 μm. B) N. lovaniensis trophozoite jammed with numerous endoparasitic stages of Pr. naegleriophila. Magnification ×16,800; bar = 1 μm. C) Enlarged detail of N. lovaniensis trophozoite with intracytoplasmic stages of Pr. naegleriophila. Some stages show binary fission indicated by the fission furrow (arrows). The endoparasites have a wrinkled gram-negative outer membrane rendering a spiny appearance to the endoparasites. Signs of damage are obvious within the cytoplasm of the host ameba. mi, mitochondria. Magnification ×43,500; bar = 0.5 μm. D) Pr. naegleriophila within vacuoles of Acanthamoebae castellanii ameba 2 days postinfection. Elementary bodies (eb) and reticulate bodies (rb) are visible. Elementary bodies harbor a smooth membrane compared with the reticulate bodies, which have a spiny shape. Magnification ×10,000; bar = 2 μm. E) Enlarged detail of A. castellanii trophozoite with intracytoplasmic stages of Pr. naegleriophila 3 days postinfection. Binary fission is observed (di). Magnification ×20,000; bar = 1 μm. F) Crescent body (arrow) within A. castellanii observed 3 days postinfection. Magnification ×20,000; bar = 1 μm.
Mentions: A. castellanii/KNic and N. lovaniensis/KNic cocultures were processed for electron microscopy as described. Ameba filled with bacteria exhibiting 3 developmental stages already described in other Parachlamydiaceae (8) were observed (Figure 1).

Bottom Line: Phenotypic, genetic, and phylogenetic analyses supported its affiliation as Protochlamydia naegleriophila sp. nov.We then developed a specific diagnostic PCR for Protochlamydia spp.When applied to bronchoalveolar lavages, results of this PCR were positive for 1 patient with pneumonia.

View Article: PubMed Central - PubMed

Affiliation: Center for Research on Intracellular Bacteria, Institute of Microbiology, University Hospital Center and University of Lausanne, Lausanne, Switzerland.

ABSTRACT
Using ameba coculture, we grew a Naegleria endosymbiont. Phenotypic, genetic, and phylogenetic analyses supported its affiliation as Protochlamydia naegleriophila sp. nov. We then developed a specific diagnostic PCR for Protochlamydia spp. When applied to bronchoalveolar lavages, results of this PCR were positive for 1 patient with pneumonia. Further studies are needed to assess the role of Protochlamydia spp. in pneumonia.

Show MeSH
Related in: MedlinePlus