Panama has a large number of wild animal species, which could host a highly diverse amount of genetic variants of Toxoplasma gondii (T. gondii). In this context, we highlight the importance of understanding the population structure of T. gondii in Panamanian wildlife and the genetic variants that can be rapidly transferred to domestic environments. This study analyzed the infection frequency and allelic composition of T. gondii in different tissue samples from wild animals. The infection frequency was measured by the PCR technique using the B1 gene as a molecular marker. The results showed a high frequency (65.6%) of infection in tissue samples collected from 221 wild animals. Stratified analyses for bird and mammal samples showed positivity rates of 67.2% and 70.12%, respectively, with no statistically significant differences. Infection frequency was also measured in five types of organs (brain, liver, heart, lung, and skeletal muscle), which showed homogeneous frequencies. The genetic diversity of the T. gondii population contained in the tissues of wild animals was analyzed by the Multilocus Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) technique, using five genes called SAG1, SAG2, SAG3, GRA6, and Apico. This analysis revealed the presence of alleles of these genes corresponding to T. gondii lineages I, II, and III. Allele III was only identified with the Apico gene in a single reptile individual analyzed. Our findings indicated diverse allelic distribution at the analyzed loci, suggesting that the tissues were probably infected by non-archetypal individuals of T. gondii.

Background: Panama is recognized as a global biodiversity hotspot; however, little is known about the genetic diversity patterns of Toxoplasma gondii in wildlife. This exploratory study evaluated the allelic and haplotypic diversity of the Alt. SAG2, GRA6, and Apico markers using wildlife tissue samples; additionally, it compared the genealogical relationships between the identified variants and a defined panel of reference sequences. Methods: Three cloned amplicons per tissue and locus were analyzed by bidirectional Sanger sequencing from 18 tissue samples collected from 13 wild animals. Locus-specific sequence relationships were examined using alignments, NeighborNet networks, and unrooted Neighbor-Joining trees based on p-distances. Results: A total of 162 clone sequences were generated, comprising 54 sequences for each locus. Alt.SAG2 variants clustered with reference alleles associated with T. gondii types II and III, whereas GRA6 variants clustered with reference alleles associated with type I, II, and III. The analyzed Apico fragment showed limited resolution among the three reference groups. Multiple sequence variants were recovered from some tissue-derived clone sets. Conclusions: The genetic variation detected in Panamanian wildlife comprises a heterogeneous set of T. gondii sequence variants associated with archetypal clonal variants and with sequence variants described in geographically distant regions. These findings suggest that the allelic variation present in the analyzed wildlife is not restricted to the regional context but is part of a broader pattern of T. gondii diversity in the Neotropics.