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Tachyzoite clones obtained from a single  Toxoplasma gondii  oocyst field sample were genotyped and characterized regarding mouse virulence. PCR-RFLP genotyping of tachyzoites initially isolated from interferon-γ-knockout (GKO) mice, BALB/c mice and VERO cell culture using the nine independent, unlinked genetic markers nSAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico revealed mixed  T. gondii  infections showing combinations of type II and type III alleles at different loci. Forty-five individual clones were obtained from all mixed  T. gondii  tachyzoite cell cultures by limiting dilution. Sixteen  T. gondii  clones showed type III alleles at all loci and 29 clones displayed a combination of type II and type III alleles at different loci. Five clone groups were identified in total, four of which include  T. gondii  clones that showed a non-canonical allele pattern and have never been described in natural infections before. All tested clones, except two, were highly virulent in BALB/c mice. The isolation of different non-canonical  T. gondii  clones originating from an oocyst sample of a single naturally infected cat demonstrate that sexual recombination as well as re-assortment of chromosomes via a sexual cross of  T. gondii  occur under natural conditions and result in the emergence of clones with increased virulence in mice.

veterinary research

Toxoplasma gondii sexual cross in a single naturally infected feline host: Generation of highly mouse-virulent and avirulent clones, genotypically different from clonal types I, II and III