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Isolation and Characterization of Two European Strains of Ehrlichia phagocytophila of Equine Origin

Department of Clinical Microbiology, Kalmar County Hospital, SE-391 85 Kalmar,¹ Department of Chemistry, Biology and Environmental Science, University of Kalmar, SE-391 82 Kalmar,³ Department of Infectious Diseases and Medical Microbiology, Lund University, SE-223 62 Lund, Sweden,² Viral and Rickettsial Zoonoses Branch, Centers for Disease Control and Prevention, Atlanta, Georgia⁴

Received 20 March 2001/ Returned for modification 7 June 2001/ Accepted 24 October 2001

We report the isolation and partial genetic characterization of two equine strains of granulocytic Ehrlichia of the genogroup Ehrlichia phagocytophila. Frozen whole-blood samples from two Swedish horses with laboratory-verified granulocytic ehrlichiosis were inoculated into HL-60 cell cultures. Granulocytic Ehrlichia was isolated and propagated from both horses. DNA extracts from the respective strains were amplified by PCR using primers directed towards the 16S rRNA gene, the groESL heat shock operon gene, and the ank gene. The amplified gene fragments were sequenced and compared to known sequences in the GenBank database. With respect to the 16S rRNA gene, the groESL gene, and the ank gene, the DNA sequences of the two equine Ehrlichia isolates were identical to sequences found in isolates from clinical cases of granulocytic ehrlichiosis in humans and domestic animals in Sweden. However, compared to amplified DNA from an American Ehrlichia strain of the E. phagocytophila genogroup, differences were found in the groESL gene and ank gene sequences.