Recent advances into the unique biology of Toxoplasma tissue cysts and the bradyzoites they house necessitates optimization of tissue cyst recovery from infected mouse brains. Here, we present data from 68 tissue cyst purifications of Type II ME49 tissue cysts in CBA/J mice performed over a period of two years. The effects if infecting with both tissue culture tachyzoites as well as ex vivo tissue cysts were assessed. Significant mortality was restricted to tachyzoite infections with female mice being more susceptible. Infection with tissue cysts was associated with both lower overall symptomology and mortality exhibiting no sex bias. Cumulatively, host sex did not impact overall tissue cyst yields, although, tachyzoite initiated infections generated significantly higher yields compared to tissue cyst-initiated infections. Notably, serial passage of tissue cysts was accompanied with a decreasing trend for subsequent cyst recovery. The time of tissue cyst harvest, a potential reflection of bradyzoite physiological state, had no significant impact on subsequent cyst yield at the selected time points. In aggregate, the data reveal the considerable heterogeneity associated with tissue cyst yield making the design of adequately powered experiments critical. This is particularly the case for drug studies where overall tissue cyst burden currently serves as the primary and often sole metric of efficacy, as the data presented here demonstrate that cyst recovery between preparations of untreated animals can mirror the reported effects of drug treatment.

Recent advances into the unique biology of Toxoplasma tissue cysts and the bradyzoites they house necessitate optimization of tissue cyst recovery from infected mouse brains. Here, we present data from 83 tissue cyst purifications of Type II ME49 tissue cysts in CBA/J mice performed over a period of 3 years.