Although the results obtained are not conclusive with regard to the pathogenic potential of this novel genus and species of Chlamydiales, we were not able to attribute the clinical signs to any known disease.
Immunohistochemical staining for chlamydia (IMAGEN Chlamydia; Oxoid, Bas ingstone, UK) showed cell-associated fluorescently stained aggregates in liver tissue, suggestive of Chlamydiales bacteria.
A PCR (4) to detect the 16S rRNA of all Chlamydiales bacteria, performed on liver tissue samples from all animals, yielded positive results in all 5 Corsican fire salamanders; in 4/7, 1/3, and 1/1 yellow spotted newts; and in 4/5 and 1/1 Strauch's spotted newts.
Despite the short sequence length of the 15 samples, the tree was well resolved with the Chlamydiales sequences and formed 3 clusters (Chlamydiaceae, Rhabdochlamydiaceae/Simkaniaceae, and Parachlamydiaceae/Waddliaceae/Criblamydiaceae) (Figure).
Molecular evidence for the existence of additional members of the order Chlamydiales.
The Chlamydiales infect a wide range of animals including humans (27,34).
Analysis of the 16S rRNA gene of microorganism WSU8-1044 from an aborted bovine foetus reveals that it is a member of the order Chlamydiales proposal of Waddliaceae ram.
For phylogenetic analyses, sequence data on complete 16S rRNA genes for each of the Chlamydiales genera were retrieved from GenBank and aligned with ClustalW (24).
in guinea-pigs and their zoonotic potential.
Phylogenetic diversity among geographically dispersed Chlamydiales endosymbionts recovered from clinical and environmental isolates of Acanthamoeba spp.
Electron micrographs of Acanthamoeba demonstrate the presence of bacteria at different developmental stages typical of the Chlamydiales, such as elementary and reticulate bodies (Figure 2).
(3,5), Bradyrhizobiaceae (37), Rickettsiales (38), Listeria monocytogenes (39), M.