Identification and genetic characterization of Toxoplasma gondii in free-ranging bristle-spined porcupine (Chaetomys subspinosus), a threatened arboreal mammal from the Brazilian Atlantic Forest

Rodrigo Alves Bezerra, Gastón Andrés Fernandez Giné, Bianca Mendes Maciel, Fernanda Amato Gaiotto, George Rêgo Albuquerque

Strains of Toxoplasma gondii in Brazil have high genetic diversity compared to North America and Europe. The bristle-spined porcupine, Chaetomys subspinosus, is often subject to hunting for human food, but it is not known whether it can be a reservoir of this parasite. The aim of this study was to verify the occurrence of T. gondii in C. subspinosus from southern Bahia, Brazil, and genetically characterize and compare the strains found with those isolated in previous studies of the same region to quantify their genetic diversity by multilocus PCR-RFLP and PCR sequencing. Twelve free-ranging C. subspinosus captured in forest fragments of the Una Biological Reserve and adjacent areas were evaluated. Three isolates of T. gondii (TgCsBr01-03) were detected. Two different genotypes were identified by applying multilocus PCR-RFLP with six molecular markers (SAG1, SAG2, SAG3, c22-8, PK1, and Apico). The isolates TgCsBr02 and TgCsBr03 were indistinguishable by this technique. However, the three isolates differed from all the reference strains and from the samples from the same region. Nevertheless, when the six genetic markers were used in multilocus PCR sequencing, all three isolates of T. gondii were different. The phylogenetic analysis revealed a greater genetic distance for TgCsBr01, which was closer to isolates from pigs from the same region, while TgCsBr02-03 was classified in the same lineage and was closer to isolates from sheep from this region. All the isolates differed from the clonal genotypes of types I, II, and III using both genotyping techniques.