Isolation and genotyping of Toxoplasma gondii in the Midwestern Brazil revealed high genetic diversity and new genotypes.

We aimed to describe the genetic diversity of Toxoplasma gondii strains isolated from domestic animals, wildlife and humans in the Midwestern Brazil. For this purpose, fragments of tissue samples (heart, brain and lung) from 35 dogs, four cats, 105 wildlife, and amniotic fluids from eight pregnant women were collected and submitted to mouse bioassay test. In a total, 22 isolates from nine dogs, one cat, ten wild animals and two women were obtained. The DNA was extracted from T. gondii isolates (lungs and brains of infected mice) and from "primary samples" (aliquots of tissue homogenate from wild animals and amniotic fluids from pregnant women) in order to screen using a Polymerase Chain Reaction (PCR) targeting a repeated 529-base pairs fragment of the T. gondii genome. All positive PCR samples were genotyped using restriction fragment length polymorphism (RFLP) analysis. To the best of our knowledge, this was the first study to report isolates of T. gondii from Leopardus pardalis, Crax fasciolata, and Dasyprocta azarae. Moreover, multilocus PCR-RFLP revealed 11 T. gondii RFLP genotypes, comprising nine previously described, including the archetypal lineage #2 type III (n = 1); two clonal Brazilian lineages, #6 type BrI (n = 1) and #8 type BrIII (n = 5); #14 (n = 2), #41 (n = 1), #108 (n = 1), #140 (n = 2), #166 (n = 4), #190 (n = 1), one potentially mixed, and two new described genotypes in two isolates. Our results confirmed the high diversity of T. gondii strains in Brazil, including identical genotypes circulating among humans, domestic dogs and wildlife.