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Applied and Environmental Microbiology, August 2006, p. 5569-5577, Vol. 72, No. 8
0099-2240/06/$08.00+0     doi:10.1128/AEM.00122-06

Isolation and Identification of Rickettsia massiliae from Rhipicephalus sanguineus Ticks Collected in Arizona

Marina E. Eremeeva,* Elizabeth A. Bosserman, Linda J. Demma, Maria L. Zambrano, Dianna M. Blau, and Gregory A. Dasch

Viral and Rickettsial Zoonoses Branch, Centers for Disease Control and Prevention, Atlanta, Georgia

Received 17 January 2006/ Accepted 21 May 2006

Twenty Rhipicephalus sanguineus ticks collected in eastern Arizona were tested by PCR assay to establish their infection rate with spotted fever group rickettsiae. With a nested PCR assay which detects a fragment of the Rickettsia genus-specific 17-kDa antigen gene (htrA), five ticks (25%) were found to contain rickettsial DNA. One rickettsial isolate was obtained from these ticks by inoculating a suspension of a triturated tick into monolayers of Vero E6 monkey kidney cells and XTC-2 clawed toad cells, and its cell culture and genotypic characteristics were determined. Fragments of the 16S rRNA, GltA, rOmpA, rOmpB, and Sca4 genes had 100%, 100%, 99%, 99%, and 99%, respectively, nucleotide similarity to Rickettsia massiliae strain Bar29, previously isolated from R. sanguineus in Catalonia, Spain (L. Beati et al., J. Clin. Microbiol. 34:2688-2694, 1996). The new isolate, AZT80, does not elicit cytotoxic effects in Vero cells and causes a persistent infection in XTC-2 cells. The AZT80 strain is susceptible to doxycycline but resistant to rifampin and erythromycin. Whether R. massiliae AZT80 is pathogenic or infectious for dogs and humans or can cause seroconversion to spotted fever group antigens in the United States is unknown.

* Corresponding author. Mailing address: Viral and Rickettsial Zoonoses Branch, Mail Stop G-13, National Center for Infectious Diseases, Centers for Disease Control and Prevention, 1600 Clifton Road NE, Atlanta, GA 30333. Phone: (404) 639-4612. Fax: (404) 639-4436. E-mail: MEremeeva{at}cdc.gov.

Applied and Environmental Microbiology, August 2006, p. 5569-5577, Vol. 72, No. 8
0099-2240/06/$08.00+0     doi:10.1128/AEM.00122-06




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