Isolation of Ehrlichia canis and its molecular and immunological analyses

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Surveys of antibody to Ehrlichia canis by immunofluorescent antibody (IFA) test and E.canis 16S rDNA by nested polymerase chain reaction (PCR) in household and stray dogs in Japan were performed. In serosurvey for E.canis infection, 17.7% (228) of 1,285 stray dogs in Okinawa Prefecture, 2.8% (11) of 399 stray dogs in Kagoshima Prefecture, 4.9% (22) of 448 private household dogs in Yamaguchi-shi, Yamaguchi Prefecture (6.2% (19) of 303 outdoor dogs were positive and all of 82 indoor dogs were negative.), and 2.3% (8) of 341 household dogs in Tokushima-shi, Tokushima Prefecture (8.3% (3) of 36 dogs infected with babesiosia gibsoni and 1.6% (5) of 305 dogs uninfected babesiosia gibsoni) were showed IFA positive.PCRs for detection of E.canis, E.muris and E.platys 16S rDNA for IFA-positive samples were performed. Three of 70 stray dogs (4.3%) in Okinawa Prefecture and 1 of 11 stray dogs (9.1%) in Kagoshima Prefecture were positive for E.canis PCR.The DNA fragments of E.muris and E.platys were not amplified. The determination of sequences for partial DNA fragments (389-bp) of 16S rDNA in four PCR-positive samples were carried out. All samples were showed quite the same sequences of the partial DNA fragments. This sequence was identical with E.canis isolated in China by Pan et al. in 2000. However, it differed from E.canis Oklahoma strain, in the United State of America, in only one base.