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Background  Obligate intracellular bacteria of arthropods often exhibit a significant role in either human health or arthropod ecology.    Methodology/Principal Findings  An obligate intracellular gamma-proteobacterium was isolated from the actively questing hard tick  Ixodes ricinus  using mammalian and amphibian cell lines. Transmission electron microscopy revealed a unique morphology of the bacterium, including intravacuolar localization of bacteria grouped predominantly in pairs and internal structures composed of electron-dense crystal-like structures and regular multilayer sheath-like structures. The isolate 20B was characterized to determine its taxonomic position using a polyphasic approach. Comparative 16S rRNA gene sequence analysis showed that this strain belongs to the family  Coxiellaceae , order  Legionellales  of Gamma-proteobacteria, and the closest relatives are different  Rickettsiella  spp. The level of 16S rRNA gene sequence similarity between strain 20B and other recognized species of the family was below 94.5%. Partial sequences of the  rpoB ,  parC  and  ftsY  genes confirmed the phylogenetic position of the new isolate. The G+C content estimated on the basis of whole genome analysis of strain 20B was 37.88%. On the basis of its phenotypic and genotypic properties, together with phylogenetic distinctiveness, we propose that strain 20B to be classified in the new genus  Diplorickettsia  as the type strain of a novel species named  Diplorickettsia massiliensis  sp. nov.    Conclusions/Significance  Considering the source of its isolation (hard tick, often biting humans) the role of this bacterium in the pathology of humans, animals and ticks should be further investigated.

A Novel Obligate Intracellular Gamma-Proteobacterium Associated with Ixodid Ticks, Diplorickettsia massiliensis, Gen. Nov., Sp. Nov