The first isolation, in vitro propagation, and genetic characterization of Ehrlichia canis in Israel

• Published in: Veterinary parasitology 1996-04, Vol.62 (3), p.331-340
• Main Authors: Keysary, Avi, Waner, Trevor, Rosner, Miri, Warner, Cynthia K., Dawson, Jacqueline E., Zass, Rosa, Biggie, Kristine L., Harrus, Shimon
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Bacterial - blood; Cell Line; DNA, Ribosomal - analysis; Dog Diseases; Dog-Rickettsia; Dogs; Ehrlichia - genetics; Ehrlichia - growth & development; Ehrlichia - isolation & purification; Ehrlichia canis; Ehrlichiosis - blood; Ehrlichiosis - diagnosis; Ehrlichiosis - veterinary; Florida; Fluorescent Antibody Technique, Indirect; Genes, Bacterial; Humans; Israel; Microscopy, Electron; Molecular Sequence Data; Monocytes - microbiology; Oklahoma; Phylogeny; Reagent Kits, Diagnostic; RNA, Ribosomal, 16S - genetics
• ISSN: 0304-4017; 1873-2550
• DOI: 10.1016/0304-4017(95)00866-7

Ehrlichia canis, the etiologic agent of canine ehrlichiosis, was isolated in Israel from a naturally infected dog with acute signs of the disease. The organism designated E. canis 611, was passaged experimentally to a beagle, from which it was propagated in primary canine monocytes. The organism was then grown in vitro in a continuous canine cell line, DH82. Nine beagles subsequently injected with whole E. canis-infected blood all developed typical symptoms of ehrlichiosis. An indirect immunofluorescence antibody test to E. canis was developed and compared with a commercial kit, revealing a good correlation between the two assays. Transmission electron microscopy of DH82 cells infected with the Israeli strain of E. canis (611), revealed organisms similar to those described in the literature: two different forms of morulae appeared, one tightly, the other loosely, packed. The 16S rRNA gene sequence obtained from the Israeli Ehrlichia isolate was compared with other isolates, E. canis Oklahoma and E. canis Florida. The Israeli strain 16S rRNA had three nucleotide differences from the Oklahoma isolate, and four nucleotide differences from the Florida isolate, in addition to one nucleotide gap in each. The Israeli isolate was found to be 0.54% different from the Oklahoma strain, and 0.61% different from the Florida strain. These are the same magnitudes of differences displayed by the other most closely related group in the phylogenetic tree, namely Ehrlichia equi, Ehrlichia phagocytophilia and the human granulocytic ehrlichia.