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Applied and Environmental Microbiology, August 2006, p. 5589-5595, Vol. 72, No. 8
0099-2240/06/$08.00+0     doi:10.1128/AEM.00532-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

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

Walairat Pornwiroon,1 Susan S. Pourciau,1 Lane D. Foil,2 and Kevin R. Macaluso1*

Department of Pathobiological Sciences, School of Veterinary Medicine, Louisiana State University, Skip Bertman Drive, SVM-3213, Baton Rouge, Louisiana 70803,1 Department of Entomology, Louisiana State University, LSB-413, Baton Rouge, Louisiana 708032

Received 6 March 2006/ Accepted 25 May 2006

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.

* Corresponding author. Mailing address: Department of Pathobiological Sciences, School of Veterinary Medicine, Louisiana State University, Skip Bertman Drive, SVM-3213, Baton Rouge, LA 70803. Phone: (225) 578-9677. Fax: (225) 578-9701. E-mail: kmacaluso{at}vetmed.lsu.edu.

Applied and Environmental Microbiology, August 2006, p. 5589-5595, Vol. 72, No. 8
0099-2240/06/$08.00+0     doi:10.1128/AEM.00532-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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