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The aim of this study was to investigate the seroprevalence and isolation of Toxoplasma gondii in free-range chickens in the state of Paraiba, northeastern Brazil. For this, blood samples were collected from 483 chickens in five municipalities in the state of Paraiba. The indirect immunofluorescence assay for anti-T. gondii antibodies was performed. The seropositive birds were slaughtered, and their brains and hearts were collected in order to perform a bioassay in mice. An epidemiological questionnaire was applied on the smallholdings visited, and univariable and multivariable logistic regressions were used to evaluate risk factors. The prevalence of chickens seropositive for T. gondii was found to be 31.5 % (152/483), and 86.1 % (56/65) of the smallholdings were positive. Among the 71 chickens subjected to bioassaying in mice, isolates of T. gondii were obtained from 33 (46.5 %). The isolates were named TgCkBrPB1 to 33. It was observed that the higher the chickens’ antibody titer was, the greater the chance of isolating the parasite also was. Sixteen of the 33 isolates (48.5 %) were lethal for all the mice inoculated until 30 days post-inoculation. The risk factors for infection with T. gondii among these free-range chickens were extensive and semi-extensive rearing systems, smallholdings located in urban areas, and presence of cats. The results indicate that the prevalence of T. gondii among chickens in the state of Paraiba is high. Many parasites remained viable in the tissues of the birds studied, and presence of the protozoan was directly related to the management of these birds.