Detection of a novel Rickettsia (Alphaproteobacteria: Rickettsiales) in rotund ticks (Ixodes kingi) from Saskatchewan, Canada

Anstead, Clare A.; Chilton, Neil B.

doi:10.1016/j.ttbdis.2012.11.013

Cited by 31 publications

(20 citation statements)

References 30 publications

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“…Total genomic DNA (gDNA) was extracted and purified from each tick using a DNeasy Blood & Tissue kit (Qiagen, Hilden, Germany), but with the modifications described previously (40,41). PCR analyses were conducted to test for the presence of Rickettsiella DNA in the total gDNA of each tick.…”

Section: Dna Extraction and Pcrmentioning

confidence: 99%

Discovery of Novel Rickettsiella spp. in Ixodid Ticks from Western Canada

Anstead

Chilton

2014

Appl Environ Microbiol

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The genomic DNA from four species of ixodid ticks in western Canada was tested for the presence of Rickettsiella by PCR analyses targeting the 16S rRNA gene. Eighty-eight percent of the Ixodes angustus (n ‫؍‬ 270), 43% of the I. sculptus (n ‫؍‬ 61), and 4% of the I. kingi (n ‫؍‬ 93) individuals examined were PCR positive for Rickettsiella, whereas there was no evidence for the presence of Rickettsiella in Dermacentor andersoni (n ‫؍‬ 45). Three different single-strand conformation polymorphism profiles of the 16S rRNA gene were detected among amplicons derived from Rickettsiella-positive ticks, each corresponding to a different sequence type. Furthermore, each sequence type was associated with a different tick species. Phylogenetic analyses of sequence data of the 16S rRNA gene and three other genes (rpsA, gidA, and sucB) revealed that all three sequence types were placed in a clade that contained species and pathotypes of the genus Rickettsiella. The bacterium in I. kingi represented the sister taxon to the Rickettsiella in I. sculptus, and both formed a clade with Rickettsiella grylli from crickets (Gryllus bimaculatus) and "R. ixodidis" from I. woodi. In contrast, the Rickettsiella in I. angustus was not a member of this clade but was placed external to the clade comprising the pathotypes of R. popilliae. The results indicate the existence of at least two new species of Rickettsiella: one in I. angustus and another in I. kingi and I. sculptus. However, the Rickettsiella strains in I. kingi and I. sculptus may also represent different species because each had unique sequences for all four genes.

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Section: Dna Extraction and Pcrmentioning

confidence: 99%

Discovery of Novel Rickettsiella spp. in Ixodid Ticks from Western Canada

Anstead

Chilton

2014

Appl Environ Microbiol

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“…There were no sequence differences among amplicons of R. peacockii for the 17-kDa gene. PCR-SSCP has been shown previously to be a powerful and effective method to prescreen for genetic variation among rickettsial DNAs derived from the total gDNA of individual ticks, particularly when combined with DNA sequencing of multiple samples of each SSCP profile type (7,15). For example, a previous study (15) demonstrated a lack of variation in the SSCP profiles of gltA amplicons derived from 386 R. peacockii-infected D. andersoni adults collected from localities up to 450 km apart (i.e., from Lethbridge, Alberta, Canada, to Outlook, SK, Canada) and among gltA amplicons derived from 66 R. montanensis-infected D. variabilis adults collected from localities up to 780 km apart (i.e., from Buffalo Pound Provincial Park, SK, Canada, to Kenora, Ontario, Canada) (15).…”

Section: Discussionmentioning

confidence: 99%

“…Total genomic DNA (gDNA) was extracted and purified from the complete bodies of 378 individual ticks using the DNeasy blood and tissue kit (Qiagen, Hilden, Germany) and the modifications described previously (7,22). All ticks were identified to the species level by morphological examination and genetic characterization (22,23), the latter of which served as a control for the quality of the gDNA extracted from each tick.…”

Section: Methodsmentioning

confidence: 99%

“…The presence/absence of Rickettsia DNA in each tick was tested by nested PCR (n-PCR) targeting part (434 bp) of the rickettsia-specific 17-kDa antigen gene using primers 17K-5 (5=-GCTTTACAAAATTCTAAA AACCATATA-3=) and 17K-3 (5=-TGTCTATCAATTCACAACTTGCC-3=) for the first phase and primers 17kD1 (5=-GCTCTTGCAACTTCTAT GTT-3=) and 17kD2 (5=-CATTGTTCGTCAGGTTGGCG-3=) for the second phase (24), as well as the protocols and cycling conditions described previously (7). All PCR-positive samples were then subjected to SSCP analyses (25) to prescreen for genetic variation.…”

Section: Methodsmentioning

confidence: 99%

“…Recently, a novel Rickettsia organism was detected in rotund ticks, Ixodes kingi, feeding on northern pocket gophers (Thomomys talpoides) in Saskatchewan, Canada (7). Phylogenetic analyses of sequence data for four genes revealed that this putative new species, provisionally named "Candidatus Rickettsia kingi," did not belong to the SFG, TRG, or TG of rickettsiae but represented a sister taxon to R. canadensis and "Candidatus Rickettsia tarasevichiae" (7), the latter of which occurs in I. persulcatus from Russia and Japan (3).…”

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confidence: 99%

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A Novel Rickettsia Species Detected in Vole Ticks (Ixodes angustus) from Western Canada

Anstead

Chilton

2013

Appl Environ Microbiol

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The genomic DNA of ixodid ticks from western Canada was tested by PCR for the presence of Rickettsia. No rickettsiae were detected in Ixodes sculptus, whereas 18% of the I. angustus and 42% of the Dermacentor andersoni organisms examined were PCR positive for Rickettsia. The rickettsiae from each tick species were characterized genetically using multiple genes. Rickettsiae within the D. andersoni organisms had sequences at four genes that matched those of R. peacockii. In contrast, the Rickettsia present within the larvae, nymphs, and adults of I. angustus had novel DNA sequences at four of the genes characterized compared to the sequences available from GenBank for all recognized species of Rickettsia and all other putative species within the genus. Phylogenetic analyses of the sequence data revealed that the rickettsiae in I. angustus do not belong to the spotted fever, transitional, or typhus groups of rickettsiae but are most closely related to "Candidatus Rickettsia kingi" and belong to a clade that also includes R. canadensis, "Candidatus Rickettsia tarasevichiae," and "Candidatus Rickettsia monteiroi."

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