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Background Currently, more than 300 genotypes of  Toxoplasma gondii  ( T. gondii)  have been described throughout the world, demonstrating its wide genetic diversity. The  SAG3  locus is one of the genes included in the genotyping panel of this parasite. It is associated with its virulence since it participates during the invasion process of the host cells. Therefore, cloning, sequencing, and bioinformatic analysis were used to deepen the understanding of the  SAG3  locus genetic diversity of  T. gondii  in blood samples from feral cats. Results Six different  SAG3  sequences were detected, five of which were detected in one feline. Three sequences were first reported here; one of them was an intragenic recombinant. In the cladogram, four out of ten  SAG3  sequences did not share nodes with others reported worldwide. Conclusions Cloning and sequencing of samples with more than one restriction pattern by PCR-RFLP were very helpful tools to demonstrate the presence of more than three genotypes of  T. gondii  in the blood of feral cats from southeastern Mexico. This suggests a potential mixed infection of multiple  T. gondii  strains and high genetic diversity of the parasites in felines in this tropical region of Mexico.

SAG3 Toxoplasma gondii cloning reveals unexpected fivefold infection in the blood of feral cats in the Mexican Caribbean