Comparative analysis of the roles of Ixodes persulcatus and I. trianguliceps ticks in natural foci of ixodid tick-borne borrelioses in the Middle Urals, Russia.

Long-term studies on natural foci of ixodid tick-borne borrelioses (ITBB) have been performed in Chusovskoi district of Perm region, the Middle Urals, where the vectors of these infections are represented by two ixodid tick species: the taiga tick Ixodes persulcatus and many times less abundant vole tick I. trianguliceps. Over 10 years, more than 6000 half-engorged ticks were collected from small forest mammals using the standard procedure, and 1027 I. persulcatus and 1142 I. trianguliceps ticks, individually or in pools, were used to inoculate BSK-2 medium. As a result, 199 Borrelia isolates were obtained. Among them, 177 isolates were identified, and the rrf(5S)-rrl(23S) intergenic spacer sequence was determined in 57 isolates. The prevalence of Borrelia infection in I. persulcatus larvae and nymphs averaged 31.0 and 53.3%, while that in I. trianguliceps larvae, nymphs, and adult ticks was five to ten times lower: 2.6, 10.2, and 8.1%, respectively. Each of the two tick species was found to carry both ITBB agents circulating in the Middle Ural foci (Borrelia garinii and B. afzelii), but the set of genogroups and genovariants of these spirochetes in I. trianguliceps proved to be far less diverse. According to the available data, this tick, compared to I. persulcatus, is generally less susceptible to Borrelia infection (especially by B. afzelii). Taking into account of its relatively low abundance, it appears that I. trianguliceps cannot seriously influence the course of epizootic process in ITBB foci of the study region, whereas highly abundant I. persulcatus with the high level of Borrelia infection is obviously a key component of these parasitic systems. A similar situation may well be typical for the entire geographic range shared by the two tick species.

Ultrastructural evidence of the ehrlichial developmental cycle in naturally infected Ixodes persulcatus ticks in the course of coinfection with Rickettsia, Borrelia, and a flavivirus.

Ehrlichiae are small gram-negative obligately intracellular bacteria that multiply within vacuoles of their host cells and are associated for a part of their life cycle with ticks, which serve as vectors for vertebrate hosts. Two morphologically and physiologically different ehrlichial cell types, reticulate cells (RC) and dense-cored cells (DC), are observed during experimental infection of cell cultures, mice, and ticks. Dense-cored cells and reticulate cells in vertebrate cell lines alternate in a developmental cycle. We observed ultrastructure of RC and DC of Ehrlichia muris in morulae in salivary gland cells and coinfection with Borrelia burgdorferi sensu lato (sl), "Candidatus Rickettsia tarasevichiae," and a flavivirus (presumably, tick-borne encephalitis virus [TBEV]) of Ixodes persulcatusticks collected in the Cis-Ural region of Russia. Polymerase chain reaction revealed 326 (81.5%) of 400 ticks carrying at least one infectious agent, and 41.5% (166 ticks) were coinfected with two to four agents. Ehrlichiae and rickettsiae were identified by sequencing of 359 bp of the 16S rRNA gene of E. muris and of 440 bp of the 16S rRNA gene and 385 bp of the gltA gene of "R. tarasevichiae." Different organs of the same tick harbored different microorganisms: TBEV in salivary gland and borreliae in midgut; E. muris in salivary gland; and "R. tarasevichiae" in midgut epithelium. Salivary gland cells contained both RC and DC, a finding that confirmed the developmental cycle in naturally infected ticks. Dense-cored cells in tick salivary glands were denser and of more irregular shape than DC in cell cultures. Ehrlichia-infected salivary gland cells had lysed cytoplasm, suggesting pathogenicity of E. muris for the tick host at the cellular level, as well as potential transmission during feeding. Rickettsiae in the midgut epithelial cells multiplied to significant numbers without altering the host cell ultrastructure. This is the first demonstration of E. muris, "R. tarasevichiae," and the ehrlichial developmental cycle in naturally infected I. persulcatus sticks.