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Abstract: Background: Bartonella is the agent of cat-scratch disease, but is also responsible for more severe conditions such as retinitis, meningoencephalitis, endocarditis and bacillary angiomatosis. Its seroprevalence is unknown in Brazil. Methods: Patients in an AIDS clinic, asymptomatic at the time of the study, were enrolled prospectively. They answered a structured questionnaire and had blood taken for serological and molecular assays. Cat breeder''s pets were tested serologically and collected ectoparasites were tested by molecular biology techniques. Blood donors, paired by age and sex, were tested for Bartonella IgG antibodies. Results: 125 HIV positive patients with a median age of 34 were studied; 61 were male and 75% were on HAART. Mean most recent CD4 count was 351–500cells/mm3. A high rate of contact with ticks, fleas and lice was observed. Bartonella IgG seroreactivity rate was 38.4% in HIV positive individuals and breeding cats was closely associated with infection (OR 3.6, CI 1.1–11.9, p <0.05). No difference was found between the sexes. Titers were 1:32 in 39 patients, 1:64 in seven, 1:128 in one and 1:256 in one. In the control group, IgG seroreactivity to Bartonella spp. was 34%, and female sex was correlated to seropositivity. Fourteen of 61 (23%) males vs 29/64 (45.3%) females were seroreactive to Bartonella (OR 2.8, CI 1.2–6.5, p <0.01). Titers were 1:32 in 29 patients, 1:64 in ten and 1:128 in four. Conclusions: Bartonella spp. seroprevalence is high in HIV positive and in blood donors in Rio de Janeiro. This may be of public health relevance. [Copyright &y& Elsevier]