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Canine bartonellosis: serological and molecular prevalence in Brazil and evidence of co-infection with Bartonella henselae and Bartonella vinsonii subsp. berkhoffii

Intracellular Pathogens Research Laboratory, Center for Comparative Medicine and Translational Research, Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, 4700 Hillsborough Street, Raleigh, NC 27606, USA.
Veterinary Research (Impact Factor: 3.38). 01/2007; 38(5):697-710. DOI: 10.1051/vetres:2007023
Source: PubMed

ABSTRACT The purpose of this study was to determine the serological and molecular prevalence of Bartonella spp. infection in a sick dog population from Brazil. At the São Paulo State University Veterinary Teaching Hospital in Botucatu, 198 consecutive dogs with clinicopathological abnormalities consistent with tick-borne infections were sampled. Antibodies to Bartonella henselae and Bartonella vinsonii subsp. berkhoffii were detected in 2.0% (4/197) and 1.5% (3/197) of the dogs, respectively. Using 16S-23S rRNA intergenic transcribed spacer (ITS) primers, Bartonella DNA was amplified from only 1/198 blood samples. Bartonella seroreactive and/or PCR positive blood samples (n=8) were inoculated into a liquid pre-enrichment growth medium (BAPGM) and subsequently sub-inoculated onto BAPGM/blood-agar plates. PCR targeting the ITS region, pap31 and rpoB genes amplified B. henselae from the blood and/or isolates of the PCR positive dog (ITS: DQ346666; pap31 gene: DQ351240; rpoB: EF196806). B. henselae and B. vinsonii subsp. berkhoffii (pap31: DQ906160; rpoB: EF196805) co-infection was found in one of the B. vinsonii subsp. berkhoffii seroreactive dogs. We conclude that dogs in this study population were infrequently exposed to or infected with a Bartonella species. The B. henselae and B. vinsonii subsp. berkhoffii strains identified in this study are genetically similar to strains isolated from septicemic cats, dogs, coyotes and human beings from other parts of the world. To our knowledge, these isolates provide the first Brazilian DNA sequences from these Bartonella species and the first evidence of Bartonella co-infection in dogs.

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    • "during three independent steps: PCR from the original specimen prior to enrichment culture, PCR from the BAPGM liquid medium after enrichment culture, and PCR from subculture agar plate isolates, if obtained, [15] [16]. Briefly, a portion of each original specimen (2 ml of aseptically-obtained ethylenediaminetetraacetic acid (EDTA)-anticoagulated blood or 200 ␮l to 2 ml of a diagnostic fluid specimen diluted proportionally to maintain a 1:5 ratio of specimen to media) was inoculated into liquid medium (BAPGM) and incubated as previously described. "
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    • "DNA was amplified by PCR targeting a variable region of the 16S-like rRNA gene [30] and H. canis DNA by a PCR targeting a fragment of the 18S rRNA gene [31], as described previously. Finally, Bartonella spp. was amplified by PCR targeting a fragment of the RNA polymerase b subunit (rpoB gene) as described elsewhere [32]. "
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    • "during three independent steps: PCR from the original specimen prior to enrichment culture, PCR from the BAPGM liquid medium after enrichment culture, and PCR from subculture agar plate isolates, if obtained, [15] [16]. Briefly, a portion of each original specimen (2 ml of aseptically-obtained ethylenediaminetetraacetic acid (EDTA)-anticoagulated blood or 200 ␮l to 2 ml of a diagnostic fluid specimen diluted proportionally to maintain a 1:5 ratio of specimen to media) was inoculated into liquid medium (BAPGM) and incubated as previously described. "
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