scholarly journals Isolation of a Spotted Fever Group Rickettsia, Rickettsia peacockii, in a Rocky Mountain Wood Tick, Dermacentor andersoni, Cell Line

2001 ◽  
Vol 67 (2) ◽  
pp. 546-552 ◽  
Author(s):  
Jason A. Simser ◽  
Ann T. Palmer ◽  
Ulrike G. Munderloh ◽  
Timothy J. Kurtti
Keyword(s):  
Cell Line ◽  
Citrate Synthase ◽  
Spotted Fever ◽  
Fluorescent Antibody ◽  
Rocky Mountain ◽  
Embryonic Cell ◽  
Host Cells ◽  
Spotted Fever Group ◽  
Dermacentor Andersoni ◽  
Rocky Mountain Wood Tick

ABSTRACT An embryonic cell line (DAE100) of the Rocky Mountain wood tick,Dermacentor andersoni, was observed by microscopy to be chronically infected with a rickettsialike organism. The organism was identified as a spotted fever group (SFG) rickettsia by PCR amplification and sequencing of portions of the 16S rRNA, citrate synthase, Rickettsia genus-specific 17-kDa antigen, and SFG-specific 190-kDa outer membrane protein A (rOmpA) genes. Sequence analysis of a partial rompA gene PCR fragment and indirect fluorescent antibody data for rOmpA and rOmpB indicated that this rickettsia was a strain (DaE100R) of Rickettsia peacockii, an SFG species presumed to be avirulent for both ticks and mammals.R. peacockii was successfully maintained in a continuous culture of DAE100 cells without apparent adverse effects on the host cells. Establishing cell lines from embryonic tissues of ticks offers an alternative technique for isolation of rickettsiae that are transovarially transmitted.

scholarly journals Rickettsia felis from Cat Fleas: Isolation and Culture in a Tick-Derived Cell Line

2006 ◽  
Vol 72 (8) ◽  
pp. 5589-5595 ◽  
Author(s):  
Walairat Pornwiroon ◽  
Susan S. Pourciau ◽  
Lane D. Foil ◽  
Kevin R. Macaluso
Keyword(s):  
Cell Line ◽  
Citrate Synthase ◽  
Ixodes Scapularis ◽  
Spotted Fever ◽  
Protein A ◽  
Etiologic Agent ◽  
Host Cells ◽  
Spotted Fever Group ◽  
Rickettsia Felis ◽  
Cat Fleas

ABSTRACT Rickettsia felis, the etiologic agent of spotted fever, is maintained in cat fleas by vertical transmission and resembles other tick-borne spotted fever group rickettsiae. In the present study, we utilized an Ixodes scapularis-derived tick cell line, ISE6, to achieve isolation and propagation of R. felis. A cytopathic effect of increased vacuolization was commonly observed in R. felis-infected cells, while lysis of host cells was not evident despite large numbers of rickettsiae. Electron microscopy identified rickettsia-like organisms in ISE6 cells, and sequence analyses of portions of the citrate synthase (gltA), 16S rRNA, Rickettsia genus-specific 17-kDa antigen, and spotted fever group-specific outer membrane protein A (ompA) genes and, notably, R. felis conjugative plasmids indicate that this cultivatable strain (LSU) was R. felis. Establishment of R. felis (LSU) in a tick-derived cell line provides an alternative and promising system for the expansion of studies investigating the interactions between R. felis and arthropod hosts.

scholarly journals Sequence and Expression Analysis of the ompA Gene of Rickettsia peacockii, an Endosymbiont of the Rocky Mountain Wood Tick, Dermacentor andersoni

2004 ◽  
Vol 70 (11) ◽  
pp. 6628-6636 ◽  
Author(s):  
Gerald D. Baldridge ◽  
Nicole Y. Burkhardt ◽  
Jason A. Simser ◽  
Timothy J. Kurtti ◽  
Ulrike G. Munderloh
Keyword(s):  
Spotted Fever ◽  
Consensus Sequence ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Rocky Mountain ◽  
Tick Population ◽  
Spotted Fever Group ◽  
East Side ◽  
Dermacentor Andersoni ◽  
Interference Phenomenon

ABSTRACT The transmission dynamics of Rocky Mountain spotted fever in Montana appears to be regulated by Rickettsia peacockii, a tick symbiotic rickettsia that interferes with transmission of virulent Rickettsia rickettsii. To elucidate the molecular relationships between the two rickettsiae and glean information on how to possibly exploit this interference phenomenon, we studied a major rickettsial outer membrane protein gene, ompA, presumed to be involved in infection and pathogenesis of spotted fever group rickettsiae (SFGR) but which is not expressed in the symbiont. Based on PCR amplification and DNA sequence analysis of the SFGR ompA gene, we demonstrate that R. peacockii is the most closely related of all known SFGR to R. rickettsii. We show that R. peacockii, originally described as East Side agent in Dermacentor andersoni ticks from the east side of the Bitterroot Valley in Montana, is still present in that tick population as well as in D. andersoni ticks collected at two widely separated locations in Colorado. The ompA genes of R. peacockii from these locations share three identical premature stop codons and a weakened ribosome binding site consensus sequence relative to ompA of R. rickettsii. The R. peacockii ompA promoter closely resembles that of R. rickettsii and is functional based on reverse transcription-PCR results. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting showed that OmpA translation products were not detected in cultured tick cells infected with R. peacockii. Double immunolabeling studies revealed actin tail structures in tick cells infected with R. rickettsii strain Hlp#2 but not in cells infected with R. peacockii.

Spotted fever group rickettsiae in immature and adult ticks (Acari: Ixodidae) from a focus of Rocky Mountain spotted fever in Connecticut

1985 ◽  
Vol 31 (12) ◽  
pp. 1131-1135 ◽  
Author(s):  
Louis A. Magnarelli ◽  
John F. Anderson ◽  
Willy Burgdorfer ◽  
Robert N. Philip ◽  
W. Adrian Chappell
Keyword(s):  
Spotted Fever ◽  
Fluorescent Antibody ◽  
Rocky Mountain ◽  
Dermacentor Variabilis ◽  
Etiologic Agent ◽  
Ixodid Ticks ◽  
Spotted Fever Group ◽  
Rocky Mountain Spotted Fever ◽  
Serum Samples ◽  
Rickettsia Rickettsii

Immature and adult ixodid ticks were collected during 1983 and 1984 in Newtown, Connecticut, an area endemic for Rocky Mountain spotted fever (RMSF), to determine prevalence of infection by spotted fever group (SFG) rickettsiae. Direct fluorescent-antibody (FA) staining revealed SFG organisms in 6 (1.8%) of 332 Dermacentor variabilis larvae, 5 (7.8%) of 64 D. variabilis nymphs, and in 2 (40%) of 5 Ixodes cookei nymphs removed from small- and medium-sized mammals. Hemolymph tests detected rickettsia-like organisms in 15 (8.8%) of 170 D. variabilis adults; 8 specimens retested by direct FA were negative. In contrast, hemocytes from 5 (8.6%) of 58 Ixodes texanus females contained organisms that stained positively in both hemolymph and direct FA tests. An indirect microimmunofluorescence test identified specific antibodies to Rickettsia rickettsii, the etiologic agent of RMSF, in serum samples from a chipmunk, raccoons, and white-footed mice. Results indicate that immature or adult ticks of at least three species may be involved in the maintenance and transmission of SFG rickettsiae at Newtown.

Experiments on Oral Secretion of the Rocky Mountain Wood Tick, Dermacentor andersoni Stiles (Acarina: Ixodidae)

1957 ◽  
Vol 89 (1) ◽  
pp. 1-5 ◽  
Author(s):  
J. D. Gregson
Keyword(s):  
British Columbia ◽  
Pacific Northwest ◽  
Rocky Mountain ◽  
Dermacentor Variabilis ◽  
Oral Secretion ◽  
Amblyomma Maculatum ◽  
Dermacentor Andersoni ◽  
The Pacific ◽  
Tick Paralysis ◽  
Rocky Mountain Wood Tick

Tick paralysis continues to be one of the most baffling and fascinating tickborne diseases in Canada. It was first reported in this country by Todd in 1912. Since then about 250 human cases, including 28 deaths, have been recorded from British Columbia. Outbreaks in cattle have affected up to 400 animals at a time, with losses in a herd as high as 65 head. Although the disease is most common in the Pacific northwest, where it is caused by the Rocky Mountain wood tick, Dermacentor andersoni Stiles, it has lately been reported as far south as Florida and has been produced by Dermacentor variabilis Say, Amblyomma maculatum Koch, and A. americanum (L.) (Gregson, 1953). The symptoms include a gradual ascending symmetrical flaccid paralysis. Apparently only man, sheep, cattle, dogs, and buffalo (one known instance) are susceptible, but even these may not necessarily be paralysed.

scholarly journals A streamlined method for transposon mutagenesis ofRickettsia parkeriyields numerous mutations that impact infection

2018 ◽  
Author(s):  
Rebecca L. Lamason ◽  
Natasha M. Kafai ◽  
Matthew D. Welch
Keyword(s):  
Host Cell ◽  
Virulence Factors ◽  
Vascular Endothelial Cells ◽  
Spotted Fever ◽  
Transposon Mutagenesis ◽  
Host Cells ◽  
Spotted Fever Group ◽  
Loss Of Function ◽  
Rickettsia Parkeri ◽  
Obligate Intracellular

AbstractThe rickettsiae are obligate intracellular alphaproteobacteria that exhibit a complex infectious life cycle in both arthropod and mammalian hosts. As obligate intracellular bacteria,Rickettsiaare highly adapted to living inside a variety of host cells, including vascular endothelial cells during mammalian infection. Although it is assumed that the rickettsiae produce numerous virulence factors that usurp or disrupt various host cell pathways, they have been challenging to genetically manipulate to identify the key bacterial factors that contribute to infection. Motivated to overcome this challenge, we sought to expand the repertoire of available rickettsial loss-of-function mutants, using an improvedmariner-based transposon mutagenesis scheme. Here, we present the isolation of over 100 transposon mutants in the spotted fever group speciesRickettsia parkeri. These mutants targeted genes implicated in a variety of pathways, including bacterial replication and metabolism, hypothetical proteins, the type IV secretion system, as well as factors with previously established roles in host cell interactions and pathogenesis. Given the need to identify critical virulence factors, forward genetic screens such as this will provide an excellent platform to more directly investigate rickettsial biology and pathogenesis.

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